D-10 Dual Program Instruction Manual English

L20012304
220-0201
D-10™
Dual Program
Instruction Manual
D-10™
Dual Program Reorder Pack
400
for HbA1c, or
200
for HbA2/F/A1c
UNITED STATES, Bio-Rad Laboratories, Inc., Hercules, CA 94547

FRANCE, Bio-Rad, Marnes-la-Coquette
Multiple-Language CD
This kit includes a multiple-language CD-ROM in the following languages: English, German, French, Spanish,
Italian, Portuguese, Swedish, Danish, and Greek.
In Vitro Diagnostic Directive (IVDD, 98/79/EC) Symbols
• Manufacturer • Authorized Representative in the • For In Vitro Diagnostic Use

• European Conformity European Union
• Hersteller • In-vitro-Diagnostikum
• EG-Konformität • Autorisierter Vertreter in der
• Fabricant • Utilisation comme test de
• Conformité européenne Europäischen Union
• Fabricante diagnostic in vitro
• Conforme a la normativa • Représentant agréé pour l'Union
• Produttore • Para uso en diagnóstico in
europea Européenne
• Fabricante vitro
• Conformità Europea • Representante Autorizado en la
• Tillverkare • Per uso diagnostico in vitro
• Conformidade com as Unión Europea
• Producent • Para uso em diagnóstico in
normas europeias • Rappresentante autorizzato per
• Κατασκευαστής vitro
• Uppfyller EU-direktiv l'Unione Europea • För in vitro-diagnostiskt bruk
• CE-mærkning • Representante Autorizado da União • Til in vitro-diagnostisk brug
• Συμμόρφωση με τα Europeia • Για in vitro διαγνωστική
ευρωπαϊκά πρότυπα • Auktoriserad EU-representant χρήση
• Autoriseret repræsentant i EU
• Εξουσιοδοτημένος αντιπρόσωπος
στην Ευρωπαϊκή Ένωση
• Use by • Lot Number • Consult Instructions for Use • Catalog Number

• Haltbar bis • Chargenbezeichnung • Gebrauchsanleitung beachten • Katalognummer
• Utiliser avant • Numéro de lot • Consulter la notice d’utilisation • Référence
• Fecha de caducidad • Número de lote • Consulte las instrucciones de uso • Número de catálogo
• Scadenza • Numero di lotto • Fare riferimento alle Istruzioni per • Numero di catalogo
• Utilizar até • Número do lote l'uso • Número de catálogo
• Användes före • Batchnummer • Consulte as instruções de utilização • Katalognummer
• Udløbsdato • Lotnummer • Se bruksanvisning • Katalognummer
• Χρήση έως • Αριθμός παρτίδας • Se brugsvejledningen • Αριθμός καταλόγου
• Συμβουλευθείτε τις οδηγίες χρήσης
• For use with • <0.05%

• Temperature Limit • Number of Tests • <0,05 %
• Zur Verwendung mit
• Temperaturgrenze • Anzahl der Tests • <0,05 %
• À utiliser avec
• Limite de température • Nombre de tests • <0,05%
• Uso con
• Límite de temperatura • Número de pruebas • <0,05%
• Da usarsi con
• Limite di temperatura • Numero di analisi • <0,05%
• Para utilizar com
• Limite de temperatura • Número de testes • <0,05 %
• För användning med
• Temperaturgränser • Antal tester • <0,05 %
• Til anvendelse sammen med
• Temperaturområde • Antal test • <0,05%
• Για χρήση με
• Όριο θερμοκρασίας • Αριθμός δοκιμασιών
• Analytical Cartridge • Calibrator Diluent • Elution Buffer 1 • Elution Buffer 2

• Analytische Kartusche • Kalibratorverdünnungslösung • Elutionspuffer 1 • Elutionspuffer 2
• Cartouche analytique • Diluant des étalons • Tampon d'élution 1 • Tampon d'élution 2
• Cartucho de análisis • Diluyente del calibrador • Tampón de elución 1 • Tampón de elución 2
• Cartuccia analitica • Diluente del calibratore • Tampone di eluizione 1 • Tampone di eluizione 2
• Coluna analítica • Diluente Calibrador • Tampão de eluição 1 • Tampão de eluição 2
• Analyskolonn • Spädningsvätska för kalibrator • Elueringsbuffert 1 • Elueringsbuffert 2
• Analysekolonne • Kalibratorfortynder • Elueringsbuffer 1 • Elueringsbuffer 2
• Αναλυτική μικροστήλη • Αραιωτικό μέσο βαθμονομητή • Ρυθμιστικό διάλυμα έκλουσης 1 • Ρυθμιστικό διάλυμα
έκλουσης 2
ii Instruction Manual L20012304

D-10™ Dual Program
In Vitro Diagnostic Directive (IVDD, 98/79/EC) Symbols
• HbA1c Calibrator Set • HbA1c Calibrator/Diluent Set • HbA1c Calibrator Level 1 • HbA1c Calibrator Level 2
• HbA1c-Kalibratorset • HbA1c-Kalibrator-/Diluentset • HbA1c-Kalibrator Level 1 • HbA1c-Kalibrator Level 2
• Gamme étalon HbA1c • Gamme étalon / diluant HbA1c • Étalon de niveau 1 HbA1c • Étalon de niveau 2 HbA1c
• Juego de calibradores para • Juego de calibradores/ • Calibrador Nivel 1 para HbA1c • Calibrador Nivel 2 para
HbA1c diluyente para HbA1c • Calibratore livello 1 dell’HbA1c HbA1c
• Set del calibratore dell’HbA1c • Set del calibratore/diluente • Calibrador Nível 1 de HbA1c • Calibratore livello 2
• Conjunto calibrador de HbA1c dell’HbA1c • HbA1c Kalibrator, nivå 1 dell’HbA1c
• HbA1c Kalibratorset • Conjunto calibrador/diluente • HbA1c kalibrator niveau 1 • Calibrador Nível 2 de HbA1c
• HbA1c kalibratorsæt de HbA1c • Βαθμονομητής HbA1c επιπέδου 1 • HbA1c Kalibrator, nivå 2
• Σετ βαθμονομητών HbA1c • HbA1c Kalibrator/spädningsset • HbA1c kalibrator niveau 2
• HbA1c kalibrator/ • Βαθμονομητής HbA1c
fortyndingssæt επιπέδου 2
• Σετ βαθμονομητών/
αραιωτικού μέσου HbA1c
• HbA2/F/A1c Calibrator Set • HbA2/F/A1c Calibrator/Diluent • HbA2/F/A1c Calibrator Level 1 • HbA2/F/A1c Calibrator Level 2
• HbA2-/F-/A1c-Kalibratorset Set • HbA2-/F-/A1c-Kalibrator Level 1 • HbA2-/F-/A1c-Kalibrator
• Gamme étalon HbA2/F/A1c • HbA2-/F-/A1c-Kalibrator-/ • Étalon de niveau 1 HbA2/F/A1c Level 2
• Juego de calibradores para Diluentset • Calibrador Nivel 1 para HbA2/F/A1c • Étalon de niveau 2
HbA2/F/A1c • Gamme étalon/diluant • Calibratore livello 1 dell’HbA2/F/A1c HbA2/F/A1c
• Set del calibratore HbA2/F/A1c • Calibrador Nível 1 de HbA2/F/A1c • Calibrador Nivel 2 para
dell’HbA2/F/A1c • Juego de calibradores/ • HbA2/F/A1c Kalibrator, nivå 1 HbA2/F/A1c
• Conjunto calibrador de diluyente para HbA2/F/A1c • HbA2/F/A1c kalibrator niveau 1 • Calibratore livello 2
HbA2/F/A1c • Set del calibratore/diluente • Βαθμονομητής HbA2/F/A1c dell’HbA2/F/A1c
• HbA2/F/A1c Kalibratorset dell’HbA2/F/A1c επιπέδου 1 • Calibrador Nível 2 de
• HbA2/F/A1c kalibratorsæt • Conjunto calibrador/diluente HbA2/F/A1c
• Σετ βαθμονομητών de HbA2/F/A1c • HbA2/F/A1c Kalibrator, nivå 2
HbA2/F/A1c • HbA2/F/A1c Kalibrator/ • HbA2/F/A1c kalibrator
spädningsset niveau 2
• HbA2/F/A1c kalibrator/ • Βαθμονομητής HbA2/F/A1c
fortyndingssæt επιπέδου 2
• Σετ βαθμονομητών/
αραιωτικού μέσου HbA2/F/A1c
• Deionized Water • Floppy Diskette • Reconstitute with • Resin

• Deionisiertes Wasser • Diskette • Rekonstitution mit • Harz
• Eau déionisée • Disquette informatique • Reconstituer avec • Résine
• Agua desionizada • Disquete • Reconstituir con • Resina
• Acqua deionizzata • Dischetto • Ricostituire con • Resina
• Água desionizada • Disquete • Reconstituir com • Resina
• Avjoniserat vatten • Diskett • Rekonstituera med • Resin
• Deioniseret vand • Diskette • Rekonstitueres med • Resin
• Απιονισμένο νερό • Δισκέτα • Ανασύσταση με • Ρητίνη
• Sample Vials • Sodium Azide • Wash/Diluent Solution • Whole Blood Primer

• Probengefäße • Natriumazid • Wasch-/Verdünnungslösung • Vollblut-Primer
• Microtubes échantillons • Azide de sodium • Solution de lavage/dilution • Sang total de
• Viales • Azida sódica • Solución de lavado/diluyente conditionnement
• Microprovette per campione • Sodio azide • Soluzione di lavaggio/diluizione • Cebador de sangre total
• Frascos de amostras • Azida de sódio • Solução de lavagem/diluente • Primer di sangue intero
• Provrör • Natriumazid • Tvätt-/spädningsvätska • Iniciador de sangue total
• Prøvekopper • Natriumazid • Vaske-/fortyndingsreagens • Helblodsprimer
• Σωληνάρια δείγματος • Αζίδιο του νατρίου • Διάλυμα έκπλυσης/αραίωσης • Fuldblodsprimer
• Εκκινητής ολικού αίματος
L20012304 Instruction Manual iii

iv Instruction Manual L20012304
Table of Contents
Intended Use .......................................................................................................................................... 2
Summary and Explanation of the Test ................................................................................................... 2
Principle of the Procedure ...................................................................................................................... 3
Kit Components ...................................................................................................................................... 3
Additional Items, Available from Bio-Rad ............................................................................................... 4
Additional Required Items, Not Available from Bio-Rad ......................................................................... 4
Precautions/Warnings............................................................................................................................. 4
Specimen Collection and Handling ........................................................................................................ 5
Preparation and Storage of Reagents .................................................................................................... 6
Indications of Instability or Deterioration of Reagents ............................................................................ 7
Procedure ............................................................................................................................................... 7
Method Selection .......................................................................................................................... 7
Installing a New Reorder Pack Lot (Update Kit Floppy Diskette) ................................................. 7
Analytical Cartridge Priming Procedure ....................................................................................... 7
Calibration for Short (HbA1c) Program .......................................................................................... 7
Routine Run for Short (HbA1c) Program ....................................................................................... 8
Calibration for Extended (HbA2/F/A1c) Program ............................................................................ 8
Routine Run for Extended (HbA2/F/A1c) Program ......................................................................... 9
Certification/Traceability to Reference Material and Method ........................................................ 9
QC Requirements ....................................................................................................................... 10
Guidelines for the Interpretation of Results .......................................................................................... 10
Limitations of the Procedure................................................................................................................. 11
Reference Values.................................................................................................................................. 12
Performance Characteristics ................................................................................................................ 13
Precision ..................................................................................................................................... 13
Accuracy ..................................................................................................................................... 14
Linearity/Recovery ...................................................................................................................... 16
Interfering Substances ................................................................................................................ 18
Sample Report Format ......................................................................................................................... 18
Product Safety Information ................................................................................................................... 41
Trademark Information.......................................................................................................................... 41
References ........................................................................................................................................... 41
L20012304 Instruction Manual 1

INTENDED USE
Short Program: The Bio-Rad D-10™ Dual Program is intended for the percent determination of hemoglobin A1c
in human whole blood using ion-exchange high-performance liquid chromatography (HPLC).
Extended Program: The Bio-Rad D-10 Dual Program is intended for the percent determination of hemoglobins
A2, F, and A1c, and for the detection of abnormal hemoglobins in human whole blood using ion-exchange
high-performance liquid chromatography (HPLC).
The Bio-Rad D-10 Dual Program is intended for Professional Use Only. For In Vitro Diagnostic Use.
SUMMARY AND EXPLANATION OF THE TEST

The D-10 Dual Program performs 400 tests for the determination of hemoglobin A1c or 200 tests for the
determination of HbA2/F/A1c.
The D-10 Dual Program consists of two different programs.
Short Program: A 3-minute program that allows the area percent determination of HbA1c only. This
program requires the use of the Dual Program HbA1c Calibrator/Diluent Set.
Extended Program: An extended 6.5-minute program that allows the area percent determination of
hemoglobins A2, F, and A1c. This program requires the use of the Dual Program
HbA2/F/A1c Calibrator/Diluent Set.
The user can switch between these programs without changing the buffers or cartridge and without
reconditioning the system.
Hemoglobin A1c – Diabetes mellitus is a condition characterized by hyperglycemia resulting from the body’s
inability to use blood glucose for energy. In Type 1 diabetes, the pancreas no longer makes insulin and
therefore, blood glucose cannot enter the cells to be used for energy. In Type 2 diabetes, either the pancreas
does not make enough insulin or the body is unable to use insulin correctly.1 The direct and indirect effects of
hyperglycemia on the human vascular system are the major source of morbidity and mortality in both Type 1 and
Type 2 diabetes. These effects include macrovascular complications (coronary artery disease, peripheral arterial
disease, and stroke) and microvascular complications (diabetic nephropathy, neuropathy, and retinopathy).2
Diabetes mellitus affects approximately 7% of the world population.3
Therapy for diabetes requires the long-term maintenance of a blood glucose level as close as possible to
a normal level, minimizing the risk of long-term vascular consequences.4,5 A single fasting blood glucose
measurement is an indication of the patient’s immediate past condition (hours), but may not represent the true
status of blood glucose regulation.6,7 The measurement of hemoglobin A1c (HbA1c) every two to three months has
been accepted as a measure of glycemic control in the care and treatment of patients with diabetes mellitus.
HbA1c, the glycohemoglobin of interest, is formed in two steps by the nonenzymatic glycation of HbA. The
first step is the formation of an unstable aldimine (labile A1c, or pre-A1c), a reversible reaction between the
carbonyl group of glucose and the N terminal valine of the β-chain of hemoglobin. Labile A1c formation is directly
proportional to the blood glucose concentration. During red blood cell circulation, some of the labile A1c is
converted (Amadori rearrangement) to form a stable ketoamine, HbA1c.8
HbA1c determination with the D-10 Dual Program has been optimized to eliminate interferences from hemoglobin
variants, labile A1c, and carbamylated hemoglobin. Please refer to Limitations of the Procedure and Performance
Characteristics for more information.
2 Instruction Manual L20012304

Hemoglobins A2 and F – A frequently occurring thalassemia, beta-thalassemia (β-thalassemia), is commonly

found in the heterozygous state as β-thalassemia minor or β-thalassemia trait. Adult blood contains primarily
hemoglobin A (HbA), a small percentage of hemoglobin A2 (HbA2), and trace amounts of fetal hemoglobin
(HbF). Carriers of β-thalassemia typically have HbA2 levels of 4–9% and HbF levels of 1–5%.9 The D-10 Dual
Program HbA2/F/A1c assay can be used for β-thalassemia screening by quantitation of HbA2 and HbF.
The most commonly occurring hemoglobin variants include hemoglobins S, E, C, and D.10 Presumptive
identification of these hemoglobin variants is made using retention time windows, such as an "S-Window" and
"C-Window."
Final determination of specific variants eluting in the windows is left to the educated judgment of the user. For
the positive confirmation of any particular hemoglobin variant, alternative separation methods are required.
PRINCIPLE OF THE PROCEDURE

The D-10 Dual Program is based on chromatographic separation of the analytes by ion-exchange
high-performance liquid chromatography (HPLC). The samples are automatically diluted on the D-10 and
injected into the analytical cartridge. The D-10 delivers a programmed buffer gradient of increasing ionic strength
to the cartridge, where the hemoglobins are separated based on their ionic interactions with the cartridge
material. The separated hemoglobins then pass through the flow cell of the filter photometer, where changes in
the absorbance at 415 nm are measured.
The D-10 software performs reduction of raw data collected from each analysis. Two-level calibration is used for
quantitation of the HbA2/F/A1c values. A sample report and a chromatogram are generated for each sample. The
A2, F, and A1c peaks are shaded. The A1c area is calculated using an exponentially modified Gaussian (EMG)
algorithm that excludes the labile A1c and carbamylated peak areas from the A1c peak area.
The D-10 Dual Program is for use only with the Bio-Rad D-10 Hemoglobin Testing System.
KIT COMPONENTS
220-0201, D-10 Dual Program Reorder Pack
The reorder pack contains supplies for 400 tests for HbA1c or 200 tests for HbA2/F/A1c :
Description
220-0210* Elution Buffer 1. Two bottles containing 2000 mL of a Bis-Tris/Phosphate buffer, pH 6.0.
Contains <0.05% sodium azide as a preservative.
220-0211* Elution Buffer 2. One bottle containing 1000 mL of a Bis-Tris/Phosphate buffer, pH 6.7.
Contains <0.05% sodium azide as a preservative.
220-0112* Wash/Diluent Solution. One bottle containing 1600 mL of deionized water with <0.05%
sodium azide as a preservative.
220-0128* HbA1c Calibrator/Diluent Set. One set consisting of 3 vials of Calibrator Level 1, 3 vials of
Calibrator Level 2, and 1 bottle of Calibrator Diluent. The calibrator vials contain lyophilized
human red blood cell hemolysate with gentamicin, tobramycin, and EDTA as preservatives.
Reconstituted volume is 7 mL per vial. Calibrator Diluent contains 100 mL deionized water
with <0.05% sodium azide as a preservative.
220-0218* HbA2/F/A1c Calibrator/Diluent Set. One set consisting of 3 vials of Calibrator Level 1,
3 vials of Calibrator Level 2, and 1 bottle of Calibrator Diluent. The calibrator vials contain
lyophilized human red blood cell hemolysate with gentamicin, tobramycin, and EDTA as
preservatives. Reconstituted volume is 7 mL per vial. Calibrator Diluent contains 100 mL
deionized water with <0.05% sodium azide as a preservative.

220-0212* Analytical Cartridge. One cation exchange cartridge, 4.0 mm ID x 30 mm.
220-0148* Whole Blood Primer. Four vials of lyophilized human red blood cell hemolysate with
gentamicin, tobramycin, and EDTA as preservatives. Reconstituted volume is 1.0 mL per
vial.
220-0215* Floppy Diskette with D-10 Dual Program parameters.
220-0149 Sample Vials. 100 polypropylene vials with pierceable caps, 1.5 mL.
Thermal Paper. One roll.
* Components are not available for individual sale.
ADDITIONAL ITEMS, AVAILABLE FROM BIO-RAD
Description
220-0297 Sample Vial Adapter, 10 x 1.5 mL
220-0375 D-10 Thermal Paper, 10 rolls
740 Lyphochek® Diabetes Bi-level Control, 6 x 0.5 mL
553 Lyphochek® Hemoglobin A2 Bi-level Control, 4 x 1.0 mL
120 Lyphochek® Hemoglobin A1c Linearity Set (1 each of 4 levels), 4 x 0.5 mL
171 Liquichek™ Diabetes Control, Level 1, 6 x 1.0 mL
172X Liquichek™ Diabetes Control, Trilevel MiniPak, 3 x 1.0 mL
ADDITIONAL REQUIRED ITEMS, NOT AVAILABLE FROM BIO-RAD

Pipettes, 5 µL, 0.5 mL, 1 mL, 7 mL
Deionized Water
Disposable Gloves
PRECAUTIONS/WARNINGS
1. For in vitro diagnostic use.
2. Wear personal protective equipment while handling all reagents and samples and while operating the
D-10 system.
3. Dispose of all waste in accordance with applicable national and/or local regulations.
4. Some reagents contain sodium azide, which may react with copper or lead plumbing to form explosive
metal azides. Use caution in disposing of these reagents. If disposing to drain, flush with large volumes
of water to prevent azide buildup.
5. Waste material containing patient samples or biological products should be considered biohazardous
when disposing or treating.
6. Chemical reagents should be handled in accordance with Good Laboratory Practices.
7. Clean up all spills immediately and thoroughly. Disinfect the area for any spills involving biohazardous
materials. Dispose of all contaminated materials appropriately.
8. Do not interchange vial or bottle caps and stoppers; this will lead to cross-contamination of reagents.
9. Never mix the contents from different bottles of the same reagent. Doing so may lead to reagent
contamination and compromise the performance of the product.

10. Each unit of whole blood used in the manufacture of the calibrators and whole blood primer was tested
by FDA accepted methods and found non-reactive for HIV-1, HIV-2, Hepatitis B (HBV), Hepatitis C
(HCV) and syphilis. No test method can offer complete assurance that products containing human
source materials will be absent of these and other infectious agents. In accordance with good laboratory
practice, all human source material should be considered potentially infectious for all infectious agents;
therefore, handle the calibrators and whole blood primer with the same precautions used with patient
specimens.
11. Adherence to the protocol specified herein is necessary to ensure proper performance of this product.
12. The calibrator vial stoppers contain dry natural rubber.
SPECIMEN COLLECTION AND HANDLING

Specimen Type
Whole blood.
Specimen Collection Precaution

Consider any materials of human origin as infectious and handle them using typical biosafety procedures.
Specimen Additives, Preservatives

The whole blood specimens should be collected in a vacuum collection tube containing EDTA.
Specimen Storage
Whole blood specimens may be stored up to 4 days at 2–8 °C or 1 day at room temperature (15–30 °C).
Specimen Preparation
1. Allow sample tubes to reach room temperature (15–30 °C) before performing the assay. No sample
preparation is required. Mixing the tubes prior to loading is not necessary. The sample tubes are loaded
into the D-10 sample rack and placed in the D-10. Ensure that the sample barcodes are facing towards
the back of the instrument. Use special rack inserts for 12, 13, and 14 mm diameter tubes. Remove all
inserts for 16 mm diameter tubes. Tubes with a height of 75 mm to 100 mm are acceptable for use.
2. If the sample is in an abnormal size/type tube, or if there is less than 2.0 mL of sample in the tube, then
the sample must be prediluted. Before pipetting, thoroughly mix the sample by gently inverting the tube.
To predilute, pipet 1.5 mL of Wash/Diluent Solution into a labeled 1.5 mL vial, followed by 5 µL of the
whole blood sample. Cap the sample vial and mix thoroughly. Use a sample vial adapter for 1.5 mL vials.
NOTE: Studies indicate that the use of MONOJECT™ ( 8881311446, Covidien, Mansfield, MA, 02048 USA)
collection tubes on the D-10 Hemoglobin Testing System can result in elevated total area. In some cases, blood
collects under the cap and causes oversampling, resulting in high total area that exceeds the acceptable range.
To use this tube type, users must either manually dilute the sample or ensure that blood does not collect under
the cap before loading the sample on the D-10 Hemoglobin Testing System. Results with total area within the
acceptable range are reportable.
Specimen Shipping
All samples of human origin must be shipped in accordance with national and international transportation
regulations.

PREPARATION AND STORAGE OF REAGENTS
Refer to the insert included with the current lot of calibrators and controls for value assignment and ranges.
When changing to a different lot of reagents and/or cartridge, the parameters from the matching floppy diskette
must be installed to ensure optimum performance of the program.
To install or change Elution Buffers, Wash/Diluent Solution, and Analytical Cartridge, follow the procedures
described in the D-10 Operation Manual, Section 4.2.
Elution Buffers and Wash/Diluent Solution

1. Allow the Elution Buffers and Wash/Diluent Solution to reach room temperature (15–30 °C) before
performing the assay. Mix each bottle by gently inverting prior to installation.
2. The Elution Buffers and Wash/Diluent Solution will be stable until the expiration date when stored
unopened at 15–30 °C. After opening the bottles, these reagents are stable for 30 days when stored at
15–30 °C.
3. With a new reorder pack, install one bottle of each reagent and follow the procedure for Installing a New
Reorder Pack Lot in the Procedure section. After 200 A1c or 100 A2/F/A1c injections, install a fresh bottle
of Elution Buffer 1. Reset the volume in the LOT INFO/Buffer 1 screen after installing this reagent.
NOTE: When using the optional D-10 Rack Loader, the two bottles of Elution Buffer 1 are installed
simultaneously. Manually resetting the volume is not required.
4. The Wash/Diluent Solution is interchangeable between Reorder Pack lots.
Whole Blood Primer
Use fresh aliquots of Whole Blood Primer when installing a new cartridge.
1. The Whole Blood Primer will be stable until the expiration date when stored unopened at 2–8 °C.
2. Prepare the Whole Blood Primer by adding 1 mL of deionized water to the vial.
3. Allow to stand for 10–15 minutes at 15–30 °C.
4. Swirl gently to dissolve and ensure complete mixing.
5. Write the reconstitution date on the label. The reconstituted Whole Blood Primer is stable for 1 day when
stored at 2–8 °C.
6. The Whole Blood Primer is interchangeable between lots.
HbA1c Calibrators
Reconstitute and store the HbA1c Calibrators as directed in the Dual Program HbA1c Calibrator/Diluent Set Insert.
HbA2/F/A1c Calibrators
Reconstitute and store the HbA2/F/A1c Calibrators as directed in the Dual Program HbA2/F/A1c Calibrator/
Diluent Set Insert.
Extracted Standards
This HPLC method does not use extracted standards.

Controls
• Reconstitute and store the Diabetes and Hemoglobin A2 controls according to the manufacturer’s
package insert.
• Bio-Rad Lyphochek Diabetes Controls and Hemoglobin A2 Controls must be diluted 1:300 prior to
analysis. Pipet 1.5 mL of Wash/Diluent Solution into a labeled 1.5 mL vial, followed by 5 µL of the
reconstituted control. Cap each control vial and mix thoroughly.
• Bio-Rad Liquichek Diabetes Controls must be diluted 1:200 prior to analysis. Pipet 1.0 mL of Wash/
Diluent Solution into a labeled 1.5 mL vial, followed by 5 µL of the control. Cap each control vial and mix
thoroughly.
INDICATIONS OF INSTABILITY OR DETERIORATION OF REAGENTS

• If reagents were frozen during shipment, mix each bottle by gently inverting before installing on
instrument.
• Do not use any reagents which have any indications of discoloration, cloudiness or precipitation.
• Do not use any reagents that show any signs of leakage.
• Do not use the calibrator or whole blood primer if the pellet is brown or the vial is broken. If the
lyophilized material contains insoluble matter, discard the material and reconstitute a new vial.
PROCEDURE
Method Selection
From the LOT INFO screen:
• Press METHOD on the touch screen.
• Select the desired method (HbA1c) or (HbA2/F/A1c).
• Press EXIT.
• Press YES to confirm the method change.
• Press EXIT.
• The selected method is indicated to the left of the status bar.
There is no need to perform a system flush unless you are installing a new lot of reagents.
Installing a New Reorder Pack Lot (Update Kit Floppy Diskette)
• Go to the LOT INFO screen.
• Press the UPDATE KIT button.
• Place the UPDATE KIT floppy diskette in the A:\drive.
• Follow the instructions on the screen to proceed with the Update Kit Procedure.
• Both the A1c and the A2/F/A1c parameters will be updated during this procedure.
• Remove the floppy diskette from the A:\drive once the procedure is completed.
Analytical Cartridge Priming Procedure
• The priming procedure must be performed once per cartridge and also after decontaminating the
D-10 System.
• The priming procedure can be performed while either the HbA1c or HbA2/F/A1c test is selected.
• Pipet 1 mL of reconstituted Whole Blood Primer into a sample vial. Put the sample vial into a sample vial
adapter labeled with a Primer barcode, then place the adapter into sample rack position 1.
• Start a run. When finished, the cartridge is ready for calibration.

Calibration for Short (HbA1c) Program
The Short (HbA1c) Program requires the use of the Dual Program HbA1c Calibrator/Diluent Set.
Calibration must be performed once, following the installation and priming of every new analytical cartridge.
Additional calibration may be performed at the discretion of the laboratory.
• Prepare samples as described in Specimen Preparation section.
• Place calibrators and controls on the D-10. Use sample vial adapters for 1.5 mL vials with calibrators and
controls. The adapters should have barcodes identifying sample type.
• Once the system is calibrated for the new cartridge, recalibration is not necessary. Recalibration is not
necessary after switching from the Extended Program.
Sample order with calibrators:
Sample # Reagent Adapter
1 HbA1c Calibrator, Level 1 Calibrator 1
2 HbA1c Calibrator, Level 2 Calibrator 2
3 Diabetes Control, Level 1 A1c Low Control
4 Diabetes Control, Level 2 A1c High Control
5–10 Patient Samples
NOTE: The analyte values printed on the calibrator reports are the calibrator assigned values. In the case of
calibration failure, the reports will indicate these assigned values; however, the statement “Calibration Failed” will
be printed at the bottom of the calibrator report. If the alert setting “Stop if calibration fails” is NOT selected (see
Section 2.4.4 of the D-10 Operation Manual), the run will continue, using the calibration factors from the previous
acceptable calibration run. For troubleshooting advice, refer to Section 6.1 of the D-10 Operation Manual.
Routine Run for Short (HbA1c) Program

Once the cartridge has been calibrated, use the following run configuration. Refer to QC Requirements for
information on control sample frequency.
Sample order without calibrators:
Sample # Reagent
1 Diabetes Control, Level 1
2 Diabetes Control, Level 2
Calibration for Extended (HbA2/F/A1c) Program

The Extended (HbA2/F/A1c) Program requires the use of the Dual Program HbA2/F/A1c Calibrator/Diluent Set.
Calibration must be performed once every 24 hours or when switching from the Short Program.
• Prepare samples as described in Specimen Preparation section.
• Place calibrators and controls on the D-10. Use sample vial adapters for 1.5 mL vials with calibrators and
controls. The adapters should have barcodes identifying sample type.

Sample order with calibrators:

Sample # Reagent Adapter
1 HbA2/F/A1c Calibrator, Level 1 Calibrator 1
2 HbA2/F/A1c Calibrator, Level 2 Calibrator 2
3 Diabetes Control, Level 1 A1c Low Control
4 Diabetes Control, Level 2 A1c High Control
5 HbA2 Control, Level 1 A2/F Low Control
6 HbA2 Control, Level 2 A2/F High Control
See previous NOTE for information regarding calibration failure.
Routine Run for Extended (HbA2/F/A1c) Program

Once the cartridge has been calibrated, use the following run configuration. Refer to QC Requirements for
information on control sample frequency.
Sample order without calibrators:
Sample # Reagent
Certification/Traceability to Reference Material and Method

The D-10 Dual Program is traceable to the reference methods of both the National Glycohemoglobin
Standardization Program (NGSP) and the International Federation of Clinical Chemistry and Laboratory
Medicine (IFCC).
The D-10 Dual Program is certified by the NGSP as having documented traceability to the Diabetes Control and
Complications Trial (DCCT) reference method. The purpose of the NGSP is to standardize glycohemoglobin test
results so that clinical laboratory results are comparable to those reported in the DCCT, where relationships to
mean blood glucose and risk for vascular complications have been established.11
The IFCC Working Group on HbA1c Standardization developed and maintains the reference measurement
procedure for HbA1c to be used as the analytical anchor for traceability of HbA1c. The IFCC reference method
separates the glycated and non-glycated N-terminal peptides of the β-chain by reversed-phase HPLC and
quantifies the analyte by mass spectrometry or capillary electrophoresis.12
The IFCC reference method is used to assign IFCC values to secondary reference materials. These materials
are used by manufacturers to assign values to product calibrators.13
HbA1c results have traditionally been reported in either conventional percent (100 × HbA1c/total Hb) or in SI
percent units (HbA1c/total Hb). In May 2007, the American Diabetes Association, European Association for the
Study of Diabetes, International Diabetes Federation, and IFCC issued a consensus statement on the worldwide
standardization of the HbA1c measurement. They recommended use of the IFCC SI units (mmol/mol).14 To
implement the use of the newly accepted SI units, the originally published master equations between the IFCC
reference and the designated comparison methods have been modified as follows:

Master Equation for Designated Comparison Method (DCM):
DCM Conversion from IFCC to DCM12

NGSP (USA) NGSP = (0.09148 × IFCC) + 2.152
Examples of Patient Results:
IFCC NGSP
43 mmol/mol 6.1%
48 mmol/mol 6.5%
53 mmol/mol 7.0%
QC Requirements
Controls should be run for the reported analyte(s).
In keeping with good laboratory practice, low and high control specimens should be included in the run once per
24 hours. A repeat run is indicated when expected control values are not obtained.
GUIDELINES FOR THE INTERPRETATION OF RESULTS

Observe the following guidelines to assure acceptable results:
1. The D-10 has passed calibration. For your reference, the slope and intercept acceptable ranges are
provided in the D-10 Dual Program Calibrator/Diluent Set Inserts.
2. Total area of each analysis should range from 1.0 to 4.0 million µvolt•second for the Extended Program
and from 1.0 to 5.0 million µvolt•second for the Short Program. Results should not be reported if the
area is outside the range.
3. The peaks are correctly identified. For your reference, the analyte retention time windows are provided in
the D-10 Dual Program Analytical Cartridge Insert.
4. Quality Control values are in range.
5. The following reportable ranges were established based on data presented in Performance
Characteristics, Linearity/Recovery.
Short Program Extended Program

Reportable Range Reportable Range
NGSP % HbA1c 3.8–18.5 3.7–18.4
IFCC mmol/mol HbA1c 18–179 17–178
% HbA2 NA 1.5–11.4
%HbF NA 0.8–16.5

When the result falls outside of the reportable range, the value is asterisked (*). HbA1c values that fall
outside of the reportable range should not be reported. To be consistent with the established reportable
ranges, HbA2 and HbF values that are lower than the reportable range should be reported as less than
1.5% for HbA2 or less than 0.8% for HbF. Very low levels of HbF may result in a negative value; these
values should be reported as less than 0.8%. Values in excess of the reportable ranges for HbA2 and
HbF should be reported as greater than 11.4% for HbA2 or greater than 16.5% for HbF.
6. Any sample with >15% or >140 mmol/mol HbA1c should be suspected of having a hemoglobin variant.15
7. Any sample with a combined area of ≥60% in the Variant (Short Program only), S, and C windows
should be suspected of having a homozygous variant or a variant–β-thalassemia phenotype. The HbA1c
result should not be reported for these samples.16
P3 Peak Resolution
The P3 peak may split due to improved resolution. When this occurs, an "Unknown" peak will be listed in the
peak table following the P3 peak (see Figure 8). The presence of this "Unknown" peak has no effect on the
HbA1c, HbA2, or HbF quantitation.
LIMITATIONS OF THE PROCEDURE

The D-10 Dual Extended Program provides an area percent determination of hemoglobins A2, F, and A1c, as
well as qualitative separation of normal and commonly occurring abnormal hemoglobins. Other less frequently
occurring variants may also elute within the established analyte identification windows.
For the positive confirmation of any particular hemoglobin variant, alternative separation methods are required.
• If a sample contains greater than 16.5% HbF, the HbF may elute in the LA1c/CHb or A1c window and
no HbF will be reported.
• Hemoglobin F quantitation may be adversely affected in the presence of LA1c greater than 2.6%.
• Elevated levels of HbA2 may be masked by concurrent iron deficiency anemia.17 In laboratory
confirmation of a β-thalassemia trait diagnosis, HbA2 levels should be considered in conjunction with
family history plus laboratory data including serum iron and iron binding capacity, red cell morphology,
hemoglobin, hematocrit, and mean corpuscular volume (MCV).17,18
• Hemoglobins D and E have been observed to coelute with HbA2. Specimens with HbA2 results greater
than 10% should be tested for the possible presence of hemoglobin variant interference.
• Neonatal samples should be tested with the Bio-Rad VARIANT™ Sickle Cell Short Program to identify
abnormal hemoglobins in newborns.
Sample Dilution
Normal total hemoglobin concentration corresponds to a total area of approximately 2.5 million µvolt • second.
Low, medium, and high whole blood patient samples were diluted to achieve total areas from 1.0 to 5.0 million
µvolt • second. These samples were run on the D-10 Dual Program to confirm that the total area does not affect
the result. The recommended total area range is from 1.0 to 4.0 million µvolt • second for the Extended Program
and from 1.0 to 5.0 million µvolt • second for the Short Program.
If the sample area is outside of the expected range, manually predilute the sample following the Specimen
Preparation guidelines.
If the sample area is still outside of the expected range, the sample should be rediluted to within the acceptable
total area count range and rerun.
NOTE: For some high total area, high HbA1c samples (e.g., 15% or 140 mmol/mol HbA1c with 4 M total area),
the A1c peak may elute outside of the established retention time window. Predilute the sample to approximately
2.5 M total area and rerun.

Abnormal Red Cell Survival
Samples from patients with hemolytic anemias will exhibit decreased glycated hemoglobin values due to the
shortened life span of the red cells. This effect will depend upon the severity of the anemia. Samples from
patients with polycythemia or post-splenectomy may exhibit increased glycated hemoglobin values due to a
somewhat longer life span of the red cells.19
Hemoglobin Variants
HbA1c values determined using the D-10 Dual Short Program for HbS trait and HbC trait specimens showed no
clinically significant difference versus values determined by an NGSP certified boronate affinity method. Typical
chromatograms for HbS and HbC trait specimens are provided in Figures 10 and 11.
HbA1c values determined using the D-10 Dual Extended Program for HbE, HbD, HbS, and HbC trait specimens
showed no clinically significant difference versus values determined by an NGSP certified boronate affinity
method. Typical chromatograms for HbE, HbD, HbS, and HbC trait specimens are provided in Figures 18–21,
respectively.
In the rare homozygous forms (SS or CC), there is no HbA present; therefore, no HbA1c value can be
determined.
A degradation peak, which coelutes with the A2 peak, may appear in HbS samples which have not been stored
appropriately (see Figure 22). This degradation peak interferes with the quantitation of HbA2. Review the A2
peak shape for all HbS samples before reporting. Do not report results if the peak shape is abnormal.
A degradation peak that interferes with the HbA1c quantitation may appear in HbE and HbD samples which have
not been stored appropriately (see Figure 23). Do not report the HbA1c result if an “Unknown” peak is identified
between the A1c and P3 peaks.
The effect of other hemoglobin variants on the quantitation of HbA1c, HbA2, and HbF has not been fully
evaluated.
Chromatograms of 4 specimens containing hemoglobin variants (as presumptively identified by gel
electrophoresis), analyzed using the Extended Program, are provided in Figures 24–27.
REFERENCE VALUES
Hemoglobin A1c Ranges20
The following HbA1c ranges may be used for interpretation of results; however, factors such as duration of
diabetes, adherence to therapy, and the age of the patient should also be considered in assessing the degree
of blood glucose control. These values are for nonpregnant individuals. "Action Suggested" depends on
individual patient circumstances. Such actions may include enhanced diabetes self-management education,
co-management with a diabetes team, referral to an endocrinologist, change in pharmacological therapy,
initiation or increased self-monitoring of blood glucose, or more frequent contact with the patient.
Hemoglobin A1c (%) Degree of Glucose Control
>8 Action Suggested†
<7 Goal‡
<6 Non-Diabetic Level
† High risk of developing long-term complications such as retinopathy, nephropathy, neuropathy and cardiopathy.
Action suggested depends on individual patient circumstances.
‡ Some danger of hypoglycemic reaction in Type I diabetics. Some glucose intolerant individuals and "sub-clinical"
diabetics may demonstrate (elevated) HbA1c in this area.

Non-Diabetic Reference Interval

The Third National Health and Nutrition Examination Survey in the U.S. included subjects 20 years of age
or older with normal fasting plasma glucose and with no previous diagnosis of diabetes. HbA1c values were
measured on the Bio-Rad DIAMAT™ System.
The weighted mean HbA1c for patients with normal fasting plasma glucose (n = 5,694) was 5.17% with a
standard deviation of 0.45%. The 95% confidence limits (mean ± 2SD) were 4.27–6.07% HbA1c.11
Since the D-10 Dual Program is certified by the NGSP, this reference interval could be used as a point of
reference until the laboratory has analyzed a sufficient number of samples to determine its own non-diabetic and
diabetic reference intervals representative of the regional population being tested.
Hemoglobin A2 and F Reference Intervals

100 samples from apparently healthy adult males Patient State HbA2 Level HbF Level
and females in North America were analyzed
using the D-10 Dual Program HbA2/F/A1c Extended Heterozygous β-thalassemia 4–9% 1–5%
Program. The mean hemoglobin A2 value was 2.6%, Normal or
Homozygous β-thalassemia 80–100%
with a 95% confidence interval of 2.2–3.1%. Increased
53 samples from apparently healthy adult males and Heterozygous HPFH* <1.5% 10–20%
females in North America were analyzed using the Homozygous HPFH* Absent 100%
D-10 Dual Program HbA2/F/A1c Extended Program. Table 1: HbA2/F Reference Intervals 9
The mean hemoglobin F value was 0.2%, with a * Hereditary Persistence of Fetal Hemoglobin
95% confidence interval of 0.0–0.8%.
Each laboratory should establish its own reference interval for interpretation of the patient report. As an initial
guideline for interpretation of results, use the published reference intervals in Table 1.
PERFORMANCE CHARACTERISTICS
Precision
The precision of the D-10 Dual Program was evaluated based on the NCCLS EP5-T2 guideline (for the Short
Program) and EP5-A guideline (for the Extended Program), "Evaluation of Precision Performance of Clinical
Chemistry Devices." In these studies, 40 runs were performed over 20 working days. In each run, aliquots
of low and high specimens were analyzed in duplicate. The results of the precision study are summarized in
Tables 2–5.
Low Patient High Patient Low Patient High Patient

Mean (% HbA1c) 5.7 9.4 Mean (% HbA1c) 5.9 13.1
Within Run (%CV) 0.8 0.5 Within Run (%CV) 0.8 0.3
Between Day (%CV) 0.7 1.0 Between Day (%CV) 1.4 0.8
Between Run (%CV) 0.5 0.5 Between Run (%CV) 0.7 0.3
Total Precision (%CV) 1.2 1.2 Total Precision (%CV) 1.8 0.9
Table 2: Precision results for HbA1c, Short Program Table 3: Precision results for HbA1c, Extended Program

Low Patient High Patient Low Patient High Patient
Mean (% HbA2) 2.2 5.4 Mean (% HbF) 2.1 8.7
Within Run (%CV) 4.5 1.7 Within Run (%CV) 2.2 1.4
Between Day (%CV) 3.4 2.7 Between Day (%CV) 1.8 1.3
Between Run (%CV) 0.0 0.0 Between Run (%CV) 1.7 0.0
Total Precision (%CV) 5.3 3.1 Total Precision (%CV) 3.3 2.0
Table 4: Precision results for HbA2, Extended Program Table 5: Precision results for HbF, Extended Program
Accuracy
D-10 Dual Extended vs Short Program
Hemoglobin A1c 14
Hemoglobin A1c results obtained

from the D-10 Dual Short and
D-10 Dual Extended Program %HbA1c
12
Extended Programs were

compared to each other. The
10
comparison study was performed
on 40 patient samples analyzed in
8
duplicate. See Figure 1.
slope = 1.0663 6
intercept = −0.5416
4
R2 = 0.9953 4 6 8 10 12 14
D-10 Dual Short Program %HbA1c
Figure 1: Correlation: D-10 Dual Extended Program (HbA1c) vs D-10 Dual

Short Program (HbA1c)

Hemoglobin A1c D-10 HbA1c vs VARIANT II HbA1c

Hemoglobin A1c results obtained from the
D-10 Dual Short and Extended Programs
were compared to values obtained on the
VARIANT™ II HbA1c Program ( 270-
2101). The comparison study was
D-10 %HbA1c
performed on 40 patient samples
analyzed in duplicate. See Figures 2 and
3.
slope = 0.9743
intercept = 0.3078
R2 = 0.9945
VARIANT II %HbA1c
Figure 2: Correlation: D-10 Dual Short Program (HbA1c) vs

VARIANT II HbA1c Program ( 270-2101)
D-10 Dual vs VARIANT II HbA1C
14
D-10 Dual Extended Program %HbA 1c
12
10
slope = 0.9906
intercept = 0.431 8
R2 = 0.9843 6
4
4 6 8 10 12 14
VARIANT II %HbA 1C
Figure 3: Correlation: D-10 Dual Extended Program (HbA1c) vs

VARIANT II HbA1c Program ( 270-2101)
Hemoglobin A2
Hemoglobin A2 results obtained from
D-10 Dual Program vs VARIANT II β-thalassemia Short Program (HbA 2)
the D-10 Dual Extended Program were 10
compared to values obtained on the 9
VARIANT II β-thalassemia Short 8
Program. The comparison study was

D-10 Dual Program %HbA2
performed on 40 patient samples 6
analyzed in duplicate. See Figure 4 for 5
results. 4
slope = 1.0898 3
2
1
R2 = 0.9832 0
0 1 2 3 4 5 6 7 8 9 10
VARIANT II %HbA2
Figure 4: Correlation: D-10 Dual Extended Program (HbA2) vs

VARIANT II β-thalassemia Short Program
Hemoglobin F
Hemoglobin F results from the D-10 D-10 Dual Program vs VARIANT II β-thalassemia Short Program (HbF)
Dual Extended Program were 16
compared to values obtained on the 14
VARIANT II β-thalassemia Short 12
Program. The comparison study was
D-10 Dual Program %HbF

10
performed on 40 patient samples
8
analyzed in duplicate. See Figure 5 for
results. 6
4
slope = 0.9497
2
0
R2 = 0.9959 0 2 4 6 8 10 12 14 16
-2
VARIANT II %HbF
Figure 5: Correlation: D-10 Dual Extended Program (HbF) vs

VARIANT II β-thalassemia Short Program
Linearity/Recovery
To demonstrate linearity on the D-10 Dual Program throughout the reportable range, low and high specimens
were derived as follows:
HbA1c (Short Program)
Low: Whole blood from a normal patient sample was supplemented with Bio-Rad Lyphochek
Hemoglobin A1c Linearity Set Level 1 to yield relative HbA1c level of 3.8%.
High: Whole blood from a diabetic patient sample was supplemented with Bio-Rad Lyphochek
HbA1c (Extended Program)
Low: Whole blood from a normal patient sample was supplemented with Bio-Rad Lyphochek
High: Whole blood from a diabetic patient sample was supplemented with Bio-Rad Lyphochek
HbA2
Low: Whole blood with a HbA2 level of 1.5%.
High: Normal whole blood spiked with purified HbA2 to a level of 11.4%.
HbF
Low: Normal whole blood with HbF level of 0.8%.
High: Normal whole blood spiked with an elevated HbF sample to a level of 16.5%.
The high specimen was diluted with the low specimen in varying ratios to yield specific relative analyte
percentages (theoretical percent). These diluted samples were analyzed with the D-10 Dual Program
(observed percent). Percent recovery was determined by dividing the observed percent by the theoretical
percent and multiplying the result by 100. Results from the Linearity Study are shown in Tables 6–9.

% Contribution
Theoretical % HbA1c Observed % HbA1c % Recovery
Low High
100 0 3.8 3.8 100
80 20 6.6 6.6 100
67 33 8.6 8.5 98.8
50 50 11.0 11.0 100
33 67 13.5 13.2 97.8
20 80 15.4 15.2 98.7
0 100 18.5 18.5 100
Table 6: Results of Study on Linearity and Recovery for HbA1c, Short Program
% Contribution
Theoretical % HbA1c Observed % HbA1c % Recovery
Low High
100 0 3.7 3.7 100
80 20 7.1 6.7 94.4
67 33 9.2 8.8 95.7
50 50 11.7 11.4 97.4
33 67 14.1 13.7 97.2
20 80 15.9 15.7 98.7
0 100 18.4 18.4 100
Table 7: Results of Study on Linearity and Recovery for HbA1c, Extended Program
% Contribution
Theoretical % HbA2 Observed % HbA2 % Recovery
Low High
100 0 1.5 1.5 100
80 20 3.5 3.3 94.3
67 33 4.9 4.7 95.9
50 50 6.5 6.4 98.5
33 67 8.2 8.1 98.8
20 80 9.5 9.6 101.1
0 100 11.4 11.4 100
Table 8: Results of Study on Linearity and Recovery for HbA2, Extended Program

% Contribution
Theoretical % HbF Observed % HbF % Recovery
Low High
100 0 0.8 0.8 100
80 20 4.0 3.7 92.5
67 33 6.1 6.0 98.4
50 50 8.7 8.8 101.1
33 67 11.3 11.4 100.9
20 80 13.4 13.6 101.5
0 100 16.5 16.5 100
Table 9: Results of Study on Linearity and Recovery for HbF, Extended Program
Interfering Substances
• Icterus, as indicated by bilirubin concentrations up to 20 mg/dL, does not interfere with the percent
determination of HbA2, HbF, or HbA1c.
• Lipemia, as indicated by triglyceride concentrations up to 5680 mg/dL, does not interfere with the
percent determination of HbA2, HbF, or HbA1c.
• Hemoglobin F concentrations up to 10% do not interfere with the percent determination of HbA1c.
• Labile A1c (LA1c/CHb-1) concentrations up to 4% do not interfere with the percent determination of
HbA1c.
• Labile A1c (LA1c/CHb-1) concentrations up to 2.6% do not interfere with the percent determination of
HbF.
• Carbamylated hemoglobin (LA1c/CHb-2) concentrations up to 3.5% do not interfere with the percent
determination of HbA1c.
SAMPLE REPORT FORMAT

Several of the following sample report examples include both IFCC (mmol/mol) and NGSP (%) units for
HbA1c. Beginning with D-10 software version 3.60, your lab's preferred reporting units for standardized HbA1c
results (i.e., NGSP only or NGSP and IFCC) can be configured in the D-10 Service Software by a Bio-Rad
representative.
NOTE: The sample report examples with the HbA1c result in % only were generated using D-10 software version
3.50.


Patient report
Bio-Rad DATE: 09/23/2010
D-10 TIME: 09:16 AM
S/N: #DA3G222606 Software version: 3.57
Sample ID: RACKB3-3-23-22-9-2010
Injection date 09/22/2010 11:45 AM
Injection #: 23 Method: HbA1c
Rack #: B3 Rack position: 3
Peak table - ID: RACKB3-3-23-22-9-2010

Peak R.time Height Area Area %
Unknown 0.14 8245 9082 0.3
A1a 0.20 6676 17769 0.5
A1b 0.28 8455 40358 1.2
F 0.44 5438 30425 0.9
LA1c/CHb-1 0.61 6180 46070 1.4
A1c 0.78 12575 125339 5.1
P3 1.36 32535 149519 4.5
A0 1.45 734587 2909750 87.4
Total Area: 3328311
Concentration: % mmol/mol
A1c 5.1 32
Figure 6: Non-Diabetic (Normal) Sample, Short Program

Patient report
D-10 TIME: 04:53 PM
Sample ID: RACK03-7-19-20-9-2010
Rack #: 03 Rack position: 7
Peak table - ID: RACK03-7-19-20-9-2010

Unknown 0.13 8151 9565 0.4
A1a 0.20 7532 21104 0.8
A1b 0.28 8670 69405 2.6
LA1c/CHb-1 0.59 9253 66063 2.5
A1c 0.77 18925 192773 9.2
P3 1.36 38585 176994 6.6
A0 1.46 530542 2149611 80.0
Total Area: 2685517
A1c 9.2 77
Figure 7: Diabetic Sample with Elevated HbA1c Level, Short Program


Control report
D-10 TIME: 04:03 PM
S/N: #DA5H505313 Software version: 3.50-1
Control ID: 33712
Rack #: D4 Rack position: 5
Peak table - ID: A1CTRH

A1a 0.21 3951 17524 1.0
A1b 0.28 5794 31276 1.8
F 0.44 1852 11829 0.7
LA1c/CHb-1 0.67 3008 18783 1.1
A1c 0.79 13142 131154 10.3
P3 1.39 14754 56407 3.2
Unknown 1.41 14990 30733 1.8
A0 1.48 382268 1445140 82.9
Total Area: 1742844
Concentration:
% A1c 10.3
Control OK
Figure 8: Split P3 Peak, Short Program

Patient report
D-10 TIME: 04:03 PM
S/N: #DA5H505327 Software version: 3.50-A1
Sample ID: 00010
Injection date 01/25/2008 03:50 PM
Rack #: C1 Rack position: 2
Peak table - ID: 00010

A1a 0.20 6503 24054 0.9
A1b 0.28 7981 47991 1.8
F 0.43 2600 16423 0.6
LA1c/CHb-1 0.63 6114 42262 1.6
LA1c/CHb-2 0.68 6802 54664 2.0
A1c 0.81 8536 109265 5.4
P3 1.36 34764 153264 5.7
A0 1.44 527758 2231123 83.3
Total Area: 2679047
Concentration:
% A1c 5.4
Figure 9: Labile A1c and Carbamylated Hemoglobin, Short Program

Patient report
D-10 TIME: 02:57 PM
Sample ID: RACKC2-1-24-17-9-2010
Peak table - ID: RACKC2-1-24-17-9-2010

Unknown 0.13 7678 9422 0.4
A1a 0.20 4591 13413 0.5
A1b 0.28 5343 43577 1.7
LA1c/CHb-1 0.68 2329 17693 0.7
A1c 0.81 7404 77290 6.3
P3 1.37 23435 104787 4.1
A0 1.48 357969 1407810 55.7
S-Window 1.67 489999 853066 33.8
Total Area: 2527057
A1c 6.3 46
Figure 10: Patient Sample with Hemoglobin S trait, Short Program

Patient report
D-10 TIME: 02:58 PM
Sample ID: RACKB3-1-13-23-9-2010
Rack #: B3 Rack position: 1
Peak table - ID: RACKB3-1-13-23-9-2010

Unknown 0.14 10208 13877 0.4
A1a 0.20 6542 23873 0.8
A1b 0.28 6573 24966 0.8
F 0.42 1810 12280 0.4
LA1c/CHb-1 0.67 2654 20186 0.7
A1c 0.79 8243 82393 5.1
P3 1.36 20991 104655 3.4
A0 1.46 470153 1979912 63.9
C-Window 1.79 407012 834752 27.0
Total Area: 3096895
A1c 5.1 32
Figure 11: Patient Sample with Hemoglobin C trait, Short Program

Patient report
D-10 TIME: 07:35 PM
S/N: #DA2D000801 Software version: 3.60
Sample ID: NP
Injection #: 23 Method: HbA2/F
Peak table - ID: NP

A1a 0.19 6956 36514 1.5
A1b 0.28 6087 24882 1.0
F 0.43 2103 12856 < 0.8 *
LA1c/CHb-1 0.69 2898 21837 0.9
A1c 0.82 8775 83537 4.8
P3 1.44 19881 146740 6.1
A0 1.69 386367 2006767 83.8
A2 3.21 3892 62165 2.7
Total Area: 2395299
F < 0.8 * ---
A1c 4.8 29
A2 2.7 ---
Figure 12: Non-Diabetic (Normal) Sample, Extended Program

Patient report
D-10 TIME: 07:35 PM
S/N: #DA2D000801 Software version: 3.60
Sample ID: DP
Peak table - ID: DP

A1a 0.20 6724 25936 1.1
A1b 0.28 7848 30791 1.4
Unknown 0.35 4181 17306 0.8
F 0.44 2263 11262 < 0.8 *
LA1c/CHb-1 0.61 8862 60275 2.6
A1c 0.80 17627 166053 9.1
P3 1.46 18062 142425 6.2
A0 1.70 352874 1768651 77.6
A2 3.22 3497 56760 2.6
Total Area: 2279461
F < 0.8 * ---
A1c 9.1 76
A2 2.6 ---
Figure 13: Diabetic Sample with Elevated HbA1c Level, Extended Program


Patient report
D-10 TIME: 12:30 PM
S/N: #DA3G222606 Software version: 3.50-A1
Sample ID: 3
Rack #: --- Rack position: 5
Peak table - ID: 3

A1a 0.20 6376 25500 1.0
A1b 0.27 8937 53433 2.2
Unknown 0.40 2943 16214 0.7
LA1c/CHb-1 0.58 8231 52350 2.1
LA1c/CHb-2 0.62 9054 58847 2.4
A1c 0.74 8890 92909 4.9
P3 1.39 21358 218178 8.9
A0 1.70 329096 1871805 76.5
A2 3.11 2838 56516 2.8
Total Area: 2445752
Concentration:
% A1c 4.9
% A2 2.8
Figure 14: Labile A1c and Carbamylated Hemoglobin, Extended Program

Patient report
D-10 TIME: 01:25 PM
Sample ID: ZX012104
Rack #: A2 Rack position: 8
Peak table - ID: ZX012104

A1a 0.20 6272 29570 2.0
A1b 0.28 4916 16182 1.1
F 0.42 1962 12076 < 0.8 *
LA1c/CHb-1 0.67 1692 11897 0.8
A1c 0.80 6280 62837 5.8
P3 1.45 5984 57425 3.9
A0 1.71 238640 1247984 85.1
A2 3.19 1647 28742 2.4
Total Area: 1466712
Concentration:
%F < 0.8 *
% A1c 5.8
% A2 2.4
Figure 15: Patient Sample with Normal Hemoglobin F and A2, Extended Program


Patient report
D-10 TIME: 01:38 PM
Sample ID: ZA013411
Rack #: A2 Rack position: 3
Peak table - ID: ZA013411

Unknown 0.14 2612 5522 0.4
A1a 0.20 3514 9401 0.7
A1b 0.28 5028 22242 1.6
F 0.43 2048 12300 < 0.8 *
LA1c/CHb-1 0.68 1612 11369 0.8
A1c 0.80 5651 55009 5.7
P3 1.45 6369 61087 4.5
A0 1.71 212830 1109235 81.4
A2 3.17 4050 75927 6.8
Total Area: 1362091
Concentration:
%F < 0.8 *
% A1c 5.7
% A2 6.8
Figure 16: Patient Sample with Elevated Hemoglobin A2, Extended Program

Patient report
D-10 TIME: 10:19 AM
S/N: #DA3C157006 Software version: 3.50-A1
Sample ID: ELEVATED_F
Peak table - ID: ELEVATED_F

A1b 0.26 13096 91428 3.4
F 0.50 26244 161932 6.1
LA1c/CHb-1 0.67 4193 22013 0.8
A1c 0.78 11032 101393 5.5
P3 1.44 10976 114651 4.3
A0 1.69 401639 2111925 79.3
A2 3.05 2775 60142 2.4
Total Area: 2663486
Concentration:
%F 6.1
% A1c 5.5
% A2 2.4
Figure 17: Patient Sample with Elevated Hemoglobin F, Extended Program


Patient report
D-10 TIME: 10:15 AM
Sample ID: AE_SAMPLE
Peak table - ID: AE_SAMPLE

Unknown 0.14 4164 8574 0.6
A1a 0.21 4678 21161 1.5
A1b 0.28 3675 15365 1.1
Unknown 0.40 2130 12344 0.9
LA1c/CHb-1 0.60 2221 18615 1.3
A1c 0.79 4059 41412 4.9
P3 1.45 11452 104181 7.4
A0 1.73 175589 927445 65.5
A2 2.87 8238 265953 22.9 *
Total Area: 1415050
Concentration:
% A1c 4.9
% A2 22.9 *
Figure 18: Patient Sample with Hemoglobin E trait, Extended Program

Patient report
D-10 TIME: 01:22 PM
Sample ID: 1
Peak table - ID: 1

Unknown 0.13 8676 25205 1.2
A1b 0.27 4387 31802 1.5
LA1c/CHb-1 0.58 2002 13534 0.6
A1c 0.76 5221 50733 5.1
P3 1.41 8821 79309 3.7
A0 1.70 185332 1096510 50.6
A2 3.05 2491 52340 2.9
Unknown 3.83 57334 772077 35.6
C-Window 4.78 11576 47243 2.2
Total Area: 2168753
Concentration:
% A1c 5.1
% A2 2.9
Figure 19: Patient Sample with Hemoglobin D trait, Extended Program

Patient report
D-10 TIME: 04:17 PM
Sample ID: RACKC1-1-29-12-10-2010
Peak table - ID: RACKC1-1-29-12-10-2010

Unknown 0.13 8504 10664 0.4
A1a 0.19 8327 34608 1.3
A1b 0.27 6796 33677 1.2
F 0.42 3538 21089 < 0.8 *
LA1c/CHb-1 0.62 3622 27045 1.0
A1c 0.79 9757 95034 6.5
P3 1.44 14953 128736 4.7
A0 1.71 261075 1494978 54.1
A2 3.15 4419 81137 3.2
S-Window 4.20 139547 836508 30.3
Total Area: 2763476
F < 0.8 * ---
A1c 6.5 48
A2 3.2 ---
Figure 20: Patient Sample with Hemoglobin S trait, Extended Program

Patient report
D-10 TIME: 04:16 PM
Sample ID: RACK08-4-47-13-10-2010

Unknown 0.13 7161 9246 0.5
A1b 0.27 5334 36345 2.0
F 0.42 1599 10302 < 0.8 *
LA1c/CHb-1 0.68 1259 8524 0.5
A1c 0.81 4248 52293 5.5
P3 1.46 6584 65969 3.6
A0 1.73 167368 1080567 58.8
A2 3.15 2517 51405 3.1
C-Window 4.77 151144 522057 28.4
Total Area: 1836709
F < 0.8 * ---
A1c 5.5 37
A2 3.1 ---
Figure 21: Patient Sample with Hemoglobin C trait, Extended Program


Patient report
D-10 TIME: 03:08 PM
S/N: #DB7G153204 Software version: 3.50-A1
Sample ID: RACK04-4-44-17-7-2008

Unknown 0.14 9359 39381 1.9
A1b 0.28 4155 11677 0.6
Unknown 0.35 3441 14338 0.7
F 0.43 2730 13843 < 0.8 *
LA1c/CHb-1 0.62 2067 17503 0.9
A1c 0.79 3895 43463 4.9
P3 1.42 23683 176599 8.6
A0 1.70 184641 903026 44.0
A2 3.09 2820 42582 2.5
Unknown 3.37 6645 121803 5.9
S-Window 4.08 120083 668153 32.6
Total Area: 2052369
Concentration:
%F < 0.8 *
% A1c 4.9
% A2 2.5
Figure 22: Patient Sample with Hemoglobin S trait, Extended Program:

Degradation Peak coeluting with HbA2 peak

Patient report
D-10 TIME: 01:54 PM
Sample ID: RACK04-6-46-24-6-2008

Unknown 0.14 7064 15847 0.5
A1b 0.28 7373 50993 1.6
F 0.43 7676 44667 1.2
LA1c/CHb-1 0.60 4330 31174 1.0
A1c 0.77 8271 75100 4.5
Unknown 1.04 2001 28237 0.9
P3 1.44 16657 149259 4.8
A0 1.68 352475 1868453 59.6
A2 2.90 25773 870334 29.6 *
Total Area: 3134064
Concentration:
%F 1.2
% A1c 4.5
% A2 29.6 *
Figure 23: Patient Sample with Hemoglobin E trait, Extended Program: Degradation
Peak (Unknown) eluting between A1c and P3 peaks


Patient report
D-10 TIME: 03:14 PM
Sample ID: SC
Peak table - ID: SC

Unknown 0.13 5052 10952 0.5
A1a 0.24 3871 16064 0.7
F 0.43 9189 46155 1.8
A0 1.77 10517 57529 2.6
A2 3.08 3567 65371 3.6
Unknown 3.34 2233 27998 1.3
S-Window 4.16 169600 1002335 45.1
Unknown 4.62 10672 40204 1.8
C-Window 4.78 408442 956829 43.0
Total Area: 2223436
Concentration:
%F 1.8
% A2 3.6
Figure 24: Patient Sample with Hemoglobin SC, Extended Program

Patient report
D-10 TIME: 11:24 AM
Sample ID: J5
Peak table - ID: J5

A1a 0.23 10456 79895 4.0
F 0.42 11598 84289 3.9
LA1c/CHb-1 0.63 7305 39230 1.9
A1c 0.72 9414 110129 7.2
P3 1.51 126044 871606 43.1
A0 1.73 166160 790097 39.1
A2 3.09 2407 45408 2.7
Total Area: 2020656
Concentration:
%F 3.9
% A1c 7.2
% A2 2.7
Figure 25: Patient Sample with Hemoglobin J-Baltimore trait, Extended Program


Patient report
D-10 TIME: 01:21 PM
Sample ID: 7
Peak table - ID: 7

A1b 0.27 6908 44576 1.4
Unknown 0.33 3482 11938 0.4
Unknown 0.40 2463 13260 0.4
LA1c/CHb-1 0.62 3100 22486 0.7
A1c 0.74 8342 85539 5.4
P3 1.39 16553 166712 5.2
A0 1.69 271633 1651430 51.4
A2 3.05 3926 85149 3.1
Unknown 3.88 5874 59984 1.9
S-Window 4.26 14982 126540 3.9
Unknown 4.57 140288 944943 29.4
Total Area: 3212557
Concentration:
% A1c 5.4
% A2 3.1
Figure 26: Patient Sample with Hemoglobin O-Arab trait, Extended Program

Patient report
D-10 TIME: 01:54 PM
Sample ID: RACK04-1-41-24-6-2008

A1a 0.16 77140 252557 9.6
Unknown 0.35 48973 215062 8.1
LA1c/CHb-1 0.59 4634 34467 1.3
A1c 0.78 6642 66960 3.5 *
P3 1.45 9312 78361 3.0
A0 1.68 384965 1969669 74.6
A2 3.15 1385 22487 1.1 *
Total Area: 2639563
Concentration:
% A1c 3.5 *
% A2 1.1 *
Figure 27: Patient Sample with Hemoglobin H trait, Extended Program

PRODUCT SAFETY INFORMATION

Whole Blood Primer, HbA1c Calibrator 1 and 2, and HbA2/F/A1c Calibrator 1 and 2
WARNING: These products contain a chemical known to the State of California to cause birth defects or
other reproductive harm. Contains <0.1% Gentamicin Sulfate and <0.1% Tobramycin.
TRADEMARK INFORMATION
D-10, DIAMAT, VARIANT, and Liquichek are trademarks of Bio-Rad Laboratories, Inc.
Lyphochek is a registered trademark of Bio-Rad Laboratories, Inc.
MONOJECT is a trademark of Covidien.
REFERENCES
1. American Diabetes Association Home Page. http://www.diabetes.org (accessed Jan 2010).
2. Fowler, M. J. Microvascular and Macrovascular Complications of Diabetes. Clin. Diabetes 2008, 26 (2),
77–82.
3. Shaw, J. E.; Sicree, R. A.; Zimmet, P. Z. Global Estimates of the Prevalence of Diabetes for 2010 and
2030. Diabetes Res. Clin. Pract. 2010, 87, 4–14.
4. Forsham, P. H. Diabetes Mellitus: A Rational Plan for Management. Postgrad. Med. 1982, 71, 139–154.
5. Hollander, P. The Case for Tight Control in Diabetes. Postgrad. Med. 1984, 75, 80–87.
6. Baynes, J. W.; Bunn, H. F.; Goldstein, D.; Harris, M.; Martin, D. B.; Peterson, C.; Winterhalter, K. National
Diabetes Data Group: Report of the Expert Committee on Glucosylated Hemoglobin. Diabetes Care
1984, 7, 602–606.
7. Nathan, D. M.; Singer, D. E.; Hurxthal, K.; Goodson, J. D. The Clinical Information Value of the
Glycosylated Hemoglobin Assay. N. Engl. J. Med. 1984, 310, 341–346.
8. Mayer, T. K.; Freedman, Z. R. Protein Glycosylation in Diabetes Mellitus: A Review of Laboratory
Measurements and of Their Clinical Utility. Clin. Chim. Acta 1983, 127, 147–184.
9. Fairbanks, V. F. Thalassemias and Related Disorders. Hemoglobinopathies and Thalassemias; Brian C.
Decker: New York, 1980; pp 18–27.
10. Fairbanks, V. F. The “Common” Hemoglobin Variants. Hemoglobinopathies and Thalassemias; Brian C.
Decker: New York, 1980; pp 10–14.
11. Rohlfing, C. L.; Little, R. R.; Wiedmeyer, H. M.; England, J. D.; Madsen, R.; Harris, M. I.; Flegal, K. M.;
Eberhardt, M. S.; Goldstein, D. E. Use of GHb (HbA1c) in Screening for Undiagnosed Diabetes in the
U.S. Population. Diabetes Care 2000, 23, 187–191.
12. International Federation of Clinical Chemistry and Laboratory Medicine Home Page. http://www.
ifcchba1c.net (accessed Jan 2010).
13. Hoelzel, W.; Weykamp, C.; Jeppsson, J. O.; Miedema, K.; Barr, J. R.; Goodall, I.; Hoshino, T.; John, W.
G.; Kobold, U.; Little, R.; Mosca, A.; Mauri, P.; Paroni, R.; Susanto, F.; Takei, I.; Thienpont, L.; Umemoto,
M.; Wiedmeyer, H. M.; IFCC Working Group on HbA1c Standardization. IFCC Reference System for
Measurement of Hemoglobin A1c in Human Blood and the National Standardization Schemes in the
United States, Japan, and Sweden: A Method-Comparison Study. Clin. Chem. 2004, 50 (1), 166–174.
14. American Diabetes Association, European Association for the Study of Diabetes, International
Federation of Clinical Chemistry and Laboratory Medicine, and International Diabetes Federation.
Consensus Statement on the Worldwide Standardization of the Hemoglobin A1c Measurement. Diabetes
Care 2007, 30 (9), 2399–2400.

15. Sacks, D. B.; Bruns, D. E.; Goldstein, D. E.; Maclaren, N. K.; McDonald, J. M.; Parrott, M. Guidelines and
Recommendations for Laboratory Analysis in the Diagnosis and Management of Diabetes Mellitus. Clin.
Chem. 2002, 48 (3), 436–472.
16. Williams, J. L. Anemias of Abnormal Globin Development—Thalassemias. In Clinical Hematology:
Principles, Procedures, Correlations, 2nd ed.; Stiene-Martin, E. A., Lotspeich-Steininger, C. A., Koepke,
J. A., Eds.; Lippincott-Raven Publishers: Philadelphia, PA, 1998; pp 217–240.
17. Rowley, P. T. The Diagnosis of Beta-Thalassemia Trait: A Review. Am. J. Hematol. 1976, 1, 129–137.
18. Ali, M. A. M.; Schwertner, E. Hemoglobin A2 Level, A Proposed Test for Confirming the Diagnosis of Iron
Deficiency. A.J.C.P. 1975, 63, 549–553.
19. Panzer, S.; Kronik, G.; Lechner, K.; Bettelheim, P.; Neumann, E.; Dudczak, R. Glycosylated Hemoglobins
(GHb): An Index of Red Cell Survival. Blood 1982, 59, 1348–1350.
20. American Diabetes Association. Standards of Medical Care for Patients with Diabetes Mellitus. Diabetes
Care 2001, 24 (Suppl. 1), 33–43.

NOTES:

TECHNICAL ASSISTANCE
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Effective Date: December 2010

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L20012304

UNITED STATES, Bio-Rad Laboratories, Inc., Hercules, CA 94547

• Manufacturer • Authorized Representative in the • For In Vitro Diagnostic Use

• Use by • Lot Number • Consult Instructions for Use • Catalog Number

• For use with • <0.05%

• Analytical Cartridge • Calibrator Diluent • Elution Buffer 1 • Elution Buffer 2

ii Instruction Manual L20012304

In Vitro Diagnostic Directive (IVDD, 98/79/EC) Symbols

• Deionized Water • Floppy Diskette • Reconstitute with • Resin

• Sample Vials • Sodium Azide • Wash/Diluent Solution • Whole Blood Primer

L20012304 Instruction Manual iii

L20012304 Instruction Manual 1

SUMMARY AND EXPLANATION OF THE TEST

2 Instruction Manual L20012304

Hemoglobins A2 and F – A frequently occurring thalassemia, beta-thalassemia (β-thalassemia), is commonly

PRINCIPLE OF THE PROCEDURE

L20012304 Instruction Manual 3

ADDITIONAL ITEMS, AVAILABLE FROM BIO-RAD

ADDITIONAL REQUIRED ITEMS, NOT AVAILABLE FROM BIO-RAD

4 Instruction Manual L20012304

SPECIMEN COLLECTION AND HANDLING

Specimen Collection Precaution

Specimen Additives, Preservatives

L20012304 Instruction Manual 5

Elution Buffers and Wash/Diluent Solution

6 Instruction Manual L20012304

INDICATIONS OF INSTABILITY OR DETERIORATION OF REAGENTS

L20012304 Instruction Manual 7

Routine Run for Short (HbA1c) Program

Calibration for Extended (HbA2/F/A1c) Program

8 Instruction Manual L20012304

Sample order with calibrators:

Routine Run for Extended (HbA2/F/A1c) Program

Certification/Traceability to Reference Material and Method

L20012304 Instruction Manual 9

DCM Conversion from IFCC to DCM12

Examples of Patient Results:

GUIDELINES FOR THE INTERPRETATION OF RESULTS

Short Program Extended Program

10 Instruction Manual L20012304

LIMITATIONS OF THE PROCEDURE

L20012304 Instruction Manual 11

12 Instruction Manual L20012304

Non-Diabetic Reference Interval

Hemoglobin A2 and F Reference Intervals

Low Patient High Patient Low Patient High Patient

L20012304 Instruction Manual 13

Hemoglobin A1c results obtained

Extended Programs were

D-10 Dual Short Program %HbA1c

Figure 1: Correlation: D-10 Dual Extended Program (HbA1c) vs D-10 Dual

14 Instruction Manual L20012304

Hemoglobin A1c D-10 HbA1c vs VARIANT II HbA1c

Figure 2: Correlation: D-10 Dual Short Program (HbA1c) vs

D-10 Dual vs VARIANT II HbA1C

Figure 3: Correlation: D-10 Dual Extended Program (HbA1c) vs

compared to values obtained on the 9

VARIANT II β-thalassemia Short 8

Program. The comparison study was

performed on 40 patient samples 6

analyzed in duplicate. See Figure 4 for 5

Figure 4: Correlation: D-10 Dual Extended Program (HbA2) vs

Dual Extended Program were 16

compared to values obtained on the 14