Principles of Medical Laboratory Science Light Sensitive Specimens

(Semi-Final) Other exceptions:  Tests:

- Biliburin
 Blood smears made from EDTA specimens must - Beta-carotene
I. SPECIMEN HANDLING & PROCESSING
be prepared within 1 hour of collection to  Use amber tube
Specimen Handling
preserve the integrity of the blood cells and  Wrap in foil
 A critical part of pre-analytic phase
prevent artifact formation due to the longed
 Proper handling from the time a specimen is collected
contact with the anticoagulant
until the test is performed helps ensure that results
 EDTA specimens for CBCs should be analyzed
obtained on the specimen accurately reflect the status of
within 6 hours but are generally stable for 24
the patient
hours at room temperature.
 CBC specimens collected in micro collection
Routine Handling
containers should be analyzed within 4 hours
 Mixing tubes by inversion  EDTA specimens for erythrocytes sedimentation
 3-10 inversions
rate (ESR) determinations must be tested within
 Helps to distribute the additive evenly while minimizing 4 hours left at room temperature or within 12
the chance of hemolysis
hours if refrigerated
 EDTA specimens for reticulocyte counts are
Hemolysis- vigorous mixing
stable up to 6 hours at room temperature and up
Microclot formation- inadequate mixing to 72 hours if refrigerated
 Glucose test specimens drawn in sodium flouride
Transporting Specimen tubes are stable for 24 hours at room temperature Specimen Processing
 Rough handling and agitation and up to 48 hours when refrigerated at 2C to 8C  Central processing area
 Hemolysis, activate platelets and break tubes  Prothrombin time (PT) results on unrefrigerated - identified
and uncentrifuged are reliable up to 24 hours - logged/accessioned
Stopper up after collection. - sorted by department
 To reduce agitation  Partial thromboplastin time (PTT) test specimens - type of processing required
 Aid clot formation in serum tubes require analysis within 4 hours of collection - evaluated for suitability for testing
 Prevent contact of the tube contents with tube stopper regardless of storage conditions
Specimen Suitability
Specimen transport bag Blood temperature specimens  Hemolysis
 Protect specimens from the effects of extreme heat or  Some specimens will precipitate or agglutinate if  Insufficient amount of specimens (QNS)
cold allowed to cool below body temperatures  Clotted
 Inadequate, inaccurate, or missing specimen ID
Delivery time limits Chilled specimens  Wrong or outdated tube
 Within 45 mins of collection  Chilling the specimen slows down metabolic  Improper handling
 Centrifuged within 1 hour of arrival in the lab  Wrong collection time
processes and protects analytes
 Prompt delivery and separation minimize the effects of  Exposure to light
metabolic processes  Delay in testing
Light sensitive specimens
 Analytes are broken down by light  Delay or error in processing
Time-limit exceptions
 “STAT” immediatly , without delay or “medical Centrifigation
Specimens requiring warmth
emergency”  Centrifuge is a machine that spins blood and other
 Testing for cold agglutins
 Must be kept at 37 degrees celsius specimens at a high number of revolutions per minute
(rpm)
 Centrifugal force created causes the cells and plasma or  Do not “pop” off using a thumb roll technique  Total protein (TP), calcium (Ca2+), and potassium (K+)
serum in blood specimens to separate concentrations are lower in capillary blood
 Specimens for tests that require serum or plasma samples
must be centrifuged Aliqout Preparation Capillary Puncture Equipment
 Transferring a portion of the specimen into one  Lancet/Incision Devices:
Tubes awaiting centrifugation or more tubes labeled with the same ID - sterile, disposable, sharp-pointed or bladed instrument
 Stoppers should remain on tubes awaiting centrifugation information as the specimen tube - punctures or cuts skin to obtain capillary blood specimen
 Can cause loss of CO2 and an increase of PH  Disposable transfer pipettes should be used when - designed for either finger or heel puncture
 Exposes the specimen to evaporation and contamination transferring serum or plasma into aliqout tubes
Laser Lancet
Evaporation II. CAPILLARY & ARTERIAL PUNCTURE  Vaporizes water in skin to produce a small hole without
 Leads to inaccurate results because of concentration of cauterizing capillaries
analytes Capillary Puncture  No risk of accidental sharps injury, no need for sharps
 Heel or finger disposal
Sources of contamination  The method of obtaining a drops of blood by
 Drop of sweat making an incision into the capillary bed in the Microcollection Containers
 Powder from gloves dermal layer of the skin  Different order of draw (CBG, EDTA..last is red)
 Small plastic tubes used to collect tiny amounts of blood
Centrifuge Operation Indicated for from capillary punctures
 Crucial that tubes to be “balanced” in a centrifuge  Infants or small children  Have color-coded bodies or stoppers & markings for
 Equal-size tubes with equal volumes of specimen  Adults and older children min/max fill levels
 The lid to the centrifuge should remain closed during
operation and should not be opened until the rotor has Not for: Microhematocrit Tubes & Sealants
come to a complete stop  Most EST or erythrocyte sedimentation rate  Capillets
methods  Disposable, narrow-bore plastic or plastic clad glass
Plasma Specimens  Coagulation studies that require plasma tubes
 May be centrifuged without delay specimens  Used primarily for hematocrit (CBC) determinations
Serum specimens  Blood cultures  One end of tube is sealed with plastic or clay sealants
 Allow to clot first  Tests that require large volumes of serum or
plasma (ex: bilirubin, liquid protime) Capillary Blood Gas Equipment (CBG)
Using a Centrifugation  CBG is only for infants
 Balance the centrifuge Principles:  CBG collection tubes- narrow-bore plastic capillary
 Set for correct speed tubes
 Close lid securely Composition of Capillary Specimens  Stirrers- (fleas) metal filings or bars inserted into tube to
 Set timer for correct time  Mixture of arterial, venous, & capillary blood mix
 Wear face shield when removing tubes  Interstitial & intracellular fluid  Magnet- used for mixing, in conjunction with stirrer
 Open lid slowly after centrifuge stops completely  More closely resembles arterial blood than  Plastic caps- used to seal tubes
venous
Stopper Removal Microscope Slides
 Removal device or robotic equipment Reference Values  Used for blood films for hematology determinations
 Wear a full-length face shield or the tube should be held  Capillary reference values may differ from Warming Devices
behind a splash shield venous values  Warming the site increases blood flow as much as 7 times
 Stopper should be covered with a gauze or tissue  Glucose concentrations are higher in capillary
 Pulled straight up and off blood
 acidosis or alkalosis  Has high oxygen content & consistency of composition
PaO2 (80-100mm Hg)  Evaluation of the
capacity of the lungs Site Selection Criteria
 How much oxygen is  Presence of collateral circulation
dissolved in the - the blood supply is from more than one artery in that area
blood - determined through Allen’s test (performed in radial artery)
 Indicates if  Artery accessibility & size
ventilation is  Type of tissue surrounding puncture site
adequate  Low risk of injuring adjacent structures or tissue during
 Decreased oxygen puncture
level in blood  Absence of inflammation, irritation, edema, hematoma,
Heel increase respiration lesion, wound, AV shunt in close proximity, or recent
Infants: rate & vise versa. arterial puncture
 Should be less than 2.0 mm PaCO2 (34-45 mm Hg)  How much carbon
 Distance between skin and bone 2.19 dioxide is dissolved Arterial Puncture Sites:
 Lateral side of heel, 3.32mm in the blood Radial Artery
 Medial 2.33 mm  Evaluates lung  Good collateral circulation
function  Easy to palpate
Finger Prick  Respiratory  Lesser chance of hematoma formation
 Depth of the incision should be less than 2.4 mm distrubances alter  Quite difficult for patients who are hypovolemic (low
 6 mos-8 years old less than 1.5 mm PaCO2 levels blood volume)
 8 years old above should be less than 2.4 mm HCO2 (22-26 mEq/L)  Measure of the  30-45 degree angle
amount of
Special Capillary Puncture Procedures bicarbonate in the Brachial Artery
 CBG - only applicable for infants blood  Large and easy to palpate
 Neonatal biliburin collection- use amber collection tube  Evaluates the  Adequate collateral circulation
to minimize the blood exposure to light bicarbonate buffer  Deeper
 Newborn/neonatal screening- detect genetic disorders system of the  Close to the basilic vein
early on kidneys  Has a nerve which can be painful or cause nerve damage
O2 saturation (97%-  Percent of oxygen when hit
Blood Smear 100%) bound to hemoglobin
 Can be done directly from capillary puncture  Determine if Femoral Artery
 First drop of blood is wiped hemoglobin is  Large and easily palpated & punctured
carrying the amount  Sometimes, only site where arterial sampling is possible
Arterial Puncture of oxygen it is  Generally used only in emergency situations or when no
 Checks respiratory function of patient capable of carrying other site is available
 To obtain blood for arterial blood gas (ABG) tests Base excess or deficit (-  Calculation of non  There’s a possibility of contamination & infection
 Arterial blood contains oxygen, venous blood contains 2) - (+2) mEq/L respiratory part of  Usually performed by physicians
carbon dioxide acid base balance on
the PaCO2, HCO2 Other sites
pH (7.35-7.45)  Measure of the and hemoglobin  In infants
acidity or alkalinity - scalp
of blood Arterial Blood - umbilical arteries
 Used to identify  Best specimen for evaluating respiratory  In adults
condition such as function - Dorsalis pedis arteries
Required Information  Thrombus formation - clot formation Special Identification System
 Patients full name  Vasovagal response - faintness or loss of  ID bracelet with self carbon adhesive label for specimen
 Medical record or identification number consciousness  Blood ID band with linear bar coded BBID #’s
 Age or date of birth  Siemens Patient Identification Check-Blood
 Room number or other patient location Sampling Errors Administration
 Date and time of test collection  Air bubbles
 Fraction of inspired oxygen (FiO2) or flow rate in liters  Delay in analysis- within 30 mins Type, Screen, and Crossmatch
per minute (L/M)  Improper mixing Anti-A - blue
 Body temperature  Improper syringe Anti-B - yellow
 Respiration rate  Obtaining venous blood by mistake Anti Rh- white
 Clinical indication for specimen collection  Use of improper anticoagulant (heparin)
 Blood drawer’s initials  Use of improper anticoagulant
 Requesting physician’s name  Use of too much (acidosis) or too little heparin Major Cross Match
(microclot)  Patient serum
Supplemental Information as Required by Institutional  Donor RBC
Policy or Regulatory Agencies Criteria for Rejection  Incubation
 Ventilation status  Air bubbles in the specimen  Wash
 Method of ventilation  Clotted specimen  Centrifuge:
 Sampling site and type of procedure  Hemolysis of the specimen (if electrolytes are - no agglutination=compatible
 Patient activity and position ordered) - agglutination = incompatible
 Working diagnosis or ICD code  Improper or absent ID or other labeling
requirements TTI (transfusion, transmissible, infection)
Materials  Improper transportation temperature (cold) - screened for HIV, hepa B, hepa C, syphilis, malaria
 Special syringe for ABG, contains lithium heparin usually  Inadequate volume of specinen for the test (QNS
in powder form or quantity not sufficient) Blood Donor Collection
 Every 1 ml of blood should have 0.05 ml of heparin  Wrong type of syringe used  General
 Antiseptic such as isopropanol or chlorhexide sponges or - Collected for transfusions, not diagnostic testing
pads for site cleaning III. Special Collections - Collected in “units” from volunteers
 Local anesthetic to numb the site (optional) Blood Bank Specimens - Requires special training & skills
 1 or 2 ml plastic syringe with a 25 or 26 gauge  Where blood products are stord and processed
 Special glass or plastic 1-5 ml self filling syringe for transfusion  Donor Eligibility
 Luer-tip normal or bubble removal cap  Yield information that determines which - Between ages 18 & 60 years (if a person is a regular donor
 Coolant, capable of maintaining the specimen at a temp products can be transfused safely into a patient he/she can exceed the 60 year old)
between 1-5 degree celsius to slow metabolism of WBC  Specimen requirement - Weight at least 110 lb
 2 by 2 inch gauze squares or pads - EDTA - Physical exam & medical history required
 Self adhering gauze bandage - Red top - Written permission from donor required
 Identification and labeling materials - Anything that involves needles (tattoo, surgeries, etc.) are not
 Puncture resistant sharps container Identification and labelling requirements eligible until a year
 Strict patient ID and specimen labelling
Hazards or Complications  Patients full name (including middle initial) Blood Donation Materials
 Arteriospasm- reflex (involuntary) contraction  Patients hospital ID no  CPD (citrate, phosphate dextrose) anticoagulant for blood
 Artery damage  Date of birth bag
 Discomfort  Date and time of collection  Sampling pouch
 Infection  Phlebotomists initials  Needle protector
 Numbness  Room number and bed number (optional)  Ultra smooth needle
 Sampling port
 RBC bag
 Plasma bag
 Platelet bag
 Cryoprecipitate bag
Red Blood Cells
 To increase the amount of rbc after trauma or surgery or
to treat severe anemia
 42 days in the refrigerator or 10 years in the freezer
Fresh Frozen Plasma
 To correct a deficiency in coagulation factors or to treat
shock due to plasma loss from burns or massive bleeding
 1 year in the freezer
Concentrate of Platelets
 To treat or prevent bleeding due to low platelet levels. To
correct functional platenet problems
 5 days at room temperature
Crypprecipitate
 To treat fibrinogen deficiencies
Autologous transfusion
 Transfusion in which you receive your owrn blood
 3 days or 1 week before your surgery
Blood Culture
 Determine presence & extent of inection
Bacteremia- presence of bacteria in the blood
Septocemia- presence of microorganism or toxin
 Identify type of organism responsible & best antibiotic to
use
 Also useful in assessing effectiveness of antibiotic
therapy
 Should be ordered on basis of patient having a condition
in which bloodstream invasion is possible and unknown
presence of fever
Specimen Requirement for Blood Culture
 2-4 blood culture sets
 Drawn 30-60 min. Apart (unless patient in critical
condition)
 Collected in special bottle, aerobic & anaerobic
 Aerobic- has air/oxygen
 Anaerobic- bacteria dies in the presence of
oxygen
 For blood culture tubes only SPS is acceptable
Skin Antisepsis: most important part of collecting
 Disinfect 3 times: alcohol, betadine (iodine),
alcohol
 Destroy skin microorganisms
 Prevent misinterpretation of microorganism as
pathogenic
 Requires 30-60 secs friction scrub
Acceptable antiseptics
 Chlorhexidine gluconate
 Tincture of iodine or povidone
Media inoculation methods
 Direct inoculation
 Collect specimen directly into blood culture
medium
 Use butterfly & specially designed holder
 Aerobic first, never push the plunger
 Syringe inoculation
 Transfer blood to bottles after draw is completed
 Safety transfer device is required
 Intermediate collection tube
 Yellow-top SPS tube
 However:
- final concentration of SPS is increased
- presents another opportunity for contamination
- increased exposure risk to lab stafff
Antimicrobial Neutralization Products
 Remove or neutralizes antimicrobials/antibiotics
from blood
 Prevent antimicrobials from inhibiting growth of
microorganisms
Coagulation Specimen
 A clear or discard tube is required
 Filled until vacuum is exhausted
 Cooling on ice during transport may be required
 Never pour 2 partially filled tubes together
2-hour Posprandial Glucose
 Excellent screening test for diabetes and other metabolic
problems
 Monitor insulin therapy
 High carbohydrate diet (2-3 days prior to test)
 Patient fasts prior to test (no eating, smoking, drinking,
other than water for at least 10 hours)
 Patient is instructed to eat special breakfast (contains 100
grams of glucose)
 Blood glucose specimen is collected 2 hours after eating
Glucose Tolerance Test (GTT)
 used to diagnose problems of carbohydrate metabolism
 For pregnant women, to confirms if there is gestational
diabetes
 Monitors patient’s tolerance to high levels of glucose
 4-5 times
 GTT preparation:
- fast at least 12 hrs but not more than 16 hrs before test
- drink water during fast & test
- do not smoke or chew gum
- juice must be finished within 5 minutes
2 hr-Lactose Tolerance Test
 Determines lack of mucosal lactase (able to breakdown
lactose into glucose or galactose)
 No mucosal lactase enzyme means you’re lactose
intolerant
 It is suggested that a 2 hour GTT be performed the day
before the LTT test
 Test is perfomered in the same manner as GTT, however
lactose is substituted for glucose
 If patient mucosal lactase, the resulting glucose curve will
be similar to the GTT curve
 “flat curve” if patient lacks the enzyme
Paternity Testing
 Requires a chain of custody protocool & specific ID
procedures
 Mother, child, & alleged father are all tested
 Blood samples are preferred, but cheek swabs are
increasing
 Blood sample testing includes ABO & Rh typing

Therapeutic Drug Monitoring

 Typically used for drugs with narrow therapeutic ranges
 Establishes & maintains drug dosage at therapeutic level
 Avoids drug toxicity
 collect blood at peak & trough levels