Alkitab University – Collage of Medical Techniques

Department of Medical Analysis3 rd. stage

Clinical Chemistry

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HbA1c Test

Written by: Supervision by:

Muhammed Raheem Hussein Dr: Sarmad N.Mageed
Sandy Adel Ishaaq
2020/06/03

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Table contents

1. Page Title
2. Table contents
3. Introduction part 1
4. Introduction part 2
5. Purpose
6. Manual Procedure
7. Advantages and disadvantage of assays for
glucose and HbA1c
8. High Hemoglobin A1c (HbA1c) Causes
9. Low Hemoglobin A1c (HbA1c) Causes
10. Reference List

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 11. Introduction

The term diabetes mellitus describes metabolic disorder with

heterogeneous aetiologies which is characterized by chronic
hyperglycaemia and disturbances of carbohydrate, fat and protein
metabolism resulting from defects in insulin secretion, insulin
action, or both. The long–term relatively specific effects of
diabetes include development of retinopathy, nephropathy and
neuropathy . People with diabetes are also at increased risk of
cardiac, peripheral arterial and cerebrovascular disease
Diabetes and lesser forms of glucose intolerance, impaired
glucose tolerance (IGT) and impaired fasting glucose (IFG), can
now be found in almost every population in the world and
epidemiological evidence suggests that, without effective
prevention and control programmes, the burden of diabetes is
likely to continue to increase globally .
Because diabetes is now affecting many in the workforce, it
has a major and deleterious impact on both individual and
national productivity. The socioeconomic consequences of
diabetes and its complications could have a seriously negative
impact on the economies of developed and developing nations
It was against this background that on 20 December, 2006,
the United Nations General Assembly unanimously passed
Resolution 61/225 declaring diabetes an international public
health issue and declaring World Diabetes Day as a United
Nations Day.

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Glycated haemoglobin (HbA1c) was initially identified as an
“unusual” haemoglobin in patients with diabetes over 40 years
ago. After that discovery, numerous small studies were
conducted correlating it to glucose measurements resulting in
the idea that HbA1c could be used as an objective measure of
glycaemic control. The A1C-Derived Average Glucose (ADAG)
study included 643 participants representing a range of A1C
levels. It established a validated relationship between A1C and
average glucose across a range of diabetes types and patient
populations . HbA1c was introduced into clinical use in the
1980s and subsequently has become a cornerstone of clinical
practice .
HbA1c reflects average plasma glucose over the previous eight
to 12 weeks . It can be performed at any time of the day
and does not require any special preparation such as fasting.
These properties have made it the preferred test for assessing
glycaemic control in people with diabetes. More recently, there
has been substantial interest in using it as a diagnostic test for
diabetes and as a screening test for persons at high risk of
diabetes
Owing in large part to the inconvenience of measuring fasting
plasma glucose levels or performing an OGTT, and day-to-day
variability in glucose, an alternative to glucose measurements for
the diagnosis of diabetes has long been sought. HbA1c has now
been recommended by an International Committee and by the
ADA as a means to diagnose diabetes .

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 Purpose
 dependence of use of glycated haemoglobin (HbA1c) for the
diagnosis of type 2 diabetes.
 Knowing how to conduct the examination in the practical
aspect
 Increasing the medical culture for those with specializing in
reading the topic.

Tools required:
-Tube - Cufette - Syringes

-Cotton - Pipette - D.W

Reagents
- Vial R1 Latex
- Vial R2a ANTIBODY
- Vial R2b ANTIBODY
- Vial R3 HAEMOLYSIS REAGENT
Devices
- Special device as Nyco Card

Specimen
- Whole blood

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Manual Procedure
Let stand reagents and specimens at room temperature. Before use, mix by
gentle swirling Latex Reagent (vial R1).

Reconstitute calibrators and controls as indicated in the insert Hemolysate

Preparation: Lyse patient’s specimen, calibrators and controls as indicated in
“Specimen Collection and Preparation”

Normal Range

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Advantages and disadvantage of assays for glucose and
HbA1c
Glucose HbA1c
Patient preparation Stringent requirements None
prior to collection if measured for
of blood diagnostic purposes.
Processing of blood Stringent requirements Avoid conditions for more
for rapid processing, than 12hr at temperatures
separation and storage >23C. Otherwise keep at
of plasma or serum 4C (stability minimally 1
minimally at 4°C. week).
Measurement Widely available Not readily available world-
wide
Standardization Standardized to Standardized to reference
reference method method procedures.
procedures.
Routine calibration Adequate Adequate
Interferences: Severe illness may Severe illness may shorten
illness increase glucose red-cell life and
concentration artifactually reduce HbA1c
values.
Haemoglobinopath No problems Most assays are not
y traits affected.
Affordability Affordable in most low Unaffordable in most low
and middle income and middle-income country
country settings. settings.

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High Hemoglobin A1c (HbA1c) Causes
 Diabetes

HbA1c ≥ 6.5% is a reliable indicator of diabetes.

Values over 5.6% indicate prediabetes.

 Being Overweight/Obese

Obesity is associated with impaired glucose tolerance and insulin resistance.

Higher body mass index (BMI), waist-to-hip ratio, and waist circumference
were all linked to significantly higher HbA1c

 Smoking
Smoking increases HbA1c levels in both diabetic and nondiabetic people

 Iron, Vitamin B12, or Folate Deficiency Anemia

Iron, vitamin B12, or folate deficiency anemia can all increase HbA1c
levels, irrespective of blood glucose levels 

The effect is dependent on the of anemia, as a study found that those with
mild anemia did not show effects on HbA1c

 Some Genetic Hemoglobin Disorders

Some genetic hemoglobin disorders can falsely increase HbA1c levels,
depending on the methods that a laboratory uses for testing 

 Alcoholism
Heavy alcohol use can falsely increase HbA1c levels. This happens because
alcohol products react with hemoglobin in the blood forming hemoglobin-
acetaldehyde (HbA1-AcH), which can be mistakenly measured as HbA1c

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Low Hemoglobin A1c (HbA1c) Causes

 Conditions that Decrease or Destroy Red Blood Cells

Conditions that decrease the number of red blood cells, such as blood loss,
donation, or transfusion can falsely decrease HbA1c levels despite possibly
elevated blood glucose.

Destruction of red blood cells (hemolysis) that can occur in infections,

autoimmune diseases, tumors, and as a side effect of some drugs also
decreases HbA1c.

 Pregnancy
HbA1c can be lower in the second trimester of pregnancy

 Some Genetic Hemoglobin Disorders

Some genetic disorders such as sickle cell anemia and thalassemia can cause
falsely low HbA1c, depending on the method the laboratory uses for the test

 Alcoholism
Excessive alcohol consumption can decrease HbA1c levels despite elevated
blood glucose because it may interfere with the binding of glucose to
hemoglobin. It also may have an effect by lowering blood glucose levels

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Reference List
1. World Health Organization. Definition, Diagnosis and Classification of Diabetes Mellitus and its
Complications. Part 1: Diagnosis and Classification of Diabetes Mellitus. WHO/NCD/NCS/99.2 ed.
Geneva, World Health Organization, 1999.

2. Hanssen KF, Bangstad HJ, Brinchmann-Hansen O et al. Blood glucose control and diabetic
microvascular complications: long-term effects of near-normoglycaemia. Diabet Med, 1992, 9:697-705

. 3. Fox CS, Coady S, Sorlie PD et al. Increasing cardiovascular disease burden due to diabetes mellitus:
the Framingham Heart Study. Circulation, 2007, 115:1544-1550.

4. Zimmet P, Alberti KG, Shaw J. Global and societal implications of the diabetes epidemic. Nature,
2001, 414:782-787.

5. Alberti KG, Zimmet P, Shaw J. International Diabetes Federation: a consensus on Type 2 diabetes
prevention. Diabet Med, 2007, 24:451-463.

6. Preventing chronic diseases: a vital investment. Geneva, World Health Organization, 2005.

7. Rahbar S, Blumenfeld O, Ranney HM. Studies of an unusual hemoglobin in patients with diabetes
mellitus. Biochem Biophys Res Commun, 1969, 36:838-843.

8. Nathan DM, Kuenen J, Borg R et al. Translating the A1C assay into estimated average glucose values.
Diabetes Care, 2008, 31:1473-1478.

9. Massi-Benedetti M. Changing targets in the treatment of type 2 diabetes. Curr Med Res Opin, 2006, 22
Suppl 2:S5-13.

10. Nathan DM, Turgeon H, Regan S. Relationship between glycated haemoglobin levels and mean
glucose levels over time. Diabetologia, 2007, 50:2239-2244

11. International Expert Committee report on the role of the A1C assay in the diagnosis of diabetes.
Diabetes Care, 2009, 32:1327-1334

12. DETECT-2 Collaboration. Is there a glycemic threshold for diabetic retinopathy? Diabetologia. In
press.

13 .Risk of progression to diabetes from prediabetes defined by HbA1c or fasting plasma glucose criteria in
Koreans.Kim CH, et al. Diabetes Res Clin Pract. 2016. PMID: 27368062

14.Christensen DL, Friis H, Mwaniki DL, et al. Prevalence of glucose intolerance and associated risk
factors in rural and urban populations of different ethnic groups in Kenya. Diabetes Res Clin
Pract. 2009;84:303–310. 

15.de Vegt F, Dekker JM, Ruhe HG, et al. Hyperglycaemia is associated with all-cause and cardiovascular
mortality in the Hoorn population: the Hoorn Study. Diabetologia. 1999;42:926–931. 

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16. Colagiuri S, Colagiuri R, Na’ati S, Muimuiheata S, Hussain Z, Palu T. The prevalence of diabetes in the
kingdom of Tonga. Diabetes Care. 2002;25:1378–1383. 

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