HANDLING AND PROCESSING OF

BLOOD SPECIMENS FOR

LABORATORY TESTING
 LEARNING OUTCOMES
• At the end of this lesson, the students must be able to:
1. Describe the steps involved in processing and
handling different types of specimens;
2. List the criteria for specimen rejection;
3. Perform routine and special specimen processing and
handling procedures;
4. Explain time constraints and exceptions for delivery
and processing of specimens; and
5. Identify Occupational Safety and Health Standards Act
(RA- 11058)- required protective equipment worn
when processing specimens.
STEPS INVOLVED IN PROCESSING AND HANDLING
DIFFERENT TYPES OF SPECIMENS
✓ The result of a test is compromised when the proper
collection procedures, storage, processing, and
transporting protocol were not followed in the
preanalytical phase.
✓ Studies show that approximately 48% to 68% of
laboratory result failures are due to prior to analysis. The
phlebotomist should be responsible enough to follow all
the appropriate steps required for each test that he is
scheduled to perform.
Routine Handling
Phlebotomist should adhere to time limits set for delivery of
specimen to the laboratory except for cases such as
emergency specimen or other conditions.
Mixing tubes by inversion - Some tests require gentle
inversion of the specimen tube to evenly distribute
additive. The number of inversion depends on the type of
additive but it is usually between 3 to 10 inversions.
Routine Handling
Phlebotomist should adhere to time limits set for delivery of
specimen to the laboratory except for cases such as
emergency specimen or other conditions.
• Transporting specimens - Handling specimen for transport
to the laboratory should be done properly because rough
handling can lead to undesirable conditions which might
hemolyze the specimen, activate platelets, affect the
coagulation, and even break the glass tube.
• The specimen should be transported with the stopper to
(1) avoid contact between contents and the stopper, (2)
minimize agitation of the specimen, and (3) aid clot
formation for serum tubes.
Special Handling
• Body temperature
1. Specimen will precipitate or agglutinate if allowed to cool below
body temperature, it should be transported at near body
temperature which is 37˚C.
2. The tubes should also be pre-warmed at 37˚C and portable heat
blocks are used during transport which could hold the temperature
for 15 minutes from removal from the incubator.
3. A heel warmer can be used for specimen that can withstand a
temperature slightly higher than 37˚C.
Examples:
Cold agglutinin, cryofibrinogen, and cryoglobulins
• Chilled specimens
Chilling slows the metabolic process which could affect the
results for some specimen. It should be completely
submerged in crushed ice and water slurry during
transport and immediately tested or refrigerated if needed.
Examples:
Adrenocorticotropic hormone (ACTH), acetone, Angiotensin-
converting enzyme (ACE), ammonia, catecholamines, free
fatty acids, gastrin, glucagon, homocysteine, lactic acid,
parathyroid hormone (PTH), ph/blood gas (if indicated),
pyruvate, renin
• Light-sensitive specimens
There are cases when exposure to light could affect the
result of a specimen, like bilirubin. The phlebotomist should
wrap the tube with aluminum foil or use light-blocking
amber-colored container.
Examples:
Bilirubin, Carotene, Red cell folate, serum folate, Vitamin B2,
Vitamin B6, Vitamin B12, Vitamin C, urine porphyrins, and
urine porphobilinogen
CRITERIA FOR SPECIMEN REJECTION
The collected specimen is transported to the central
processing or triage for screening and prioritizing. The
specimen are:
(1) identified,
(2) logged or accessioned,
(3) sorted by department and type of processing, and
(4) evaluated for specimen suitability which is necessary for
accurate results.
The specimen is rejected if it did not meet a specific criterion
such as:
o Specimen is not identified properly.
o It has inadequate volume to complete the test.
o Hemolysis
o The use of wrong tube for collection.
o Outdated tube is used
o Improper handling (improper mixing)
o Contaminated specimen
o Insufficient specimen or "Quality Not Sufficient" (QNS)
o Incorrect collection time
o Exposure to light
o Procedure did not follow testing time limits
o Delay or error in processing
TIME CONSTRAINTS AND EXCEPTIONS FOR DELIVERY AND
PROCESSING OF SPECIMENS

Delivery time limits

✓ The specimen should be transported to the laboratory immediately
after collection. Routine blood specimen is expected to reach the
laboratory within 45 minutes.
✓ For specimen that needs centrifugation, it should be done in 1 hour.
✓ Hematology specimen with EDTA which are placed in tubes with
lavender or purple stopper should not be centrifuged.
Time Limit Exceptions
The delivery time limit has some exception such as specimen that
are marked as "STAT" or "emergency", it takes priority over all other
specimen in terms of transportation, processing and testing. Other
exceptions to the time limit rule are as follows:
1. Blood smear from EDTA specimen - prepared 1 hour from collection
2. EDTA specimen for CBC - analyzed within 6 hours but are stable 24
hours from collected if stored in room temperature.
3. EDTA specimen for erythrocyte sedimentation rate (ESR) determination -
tested 4 hours (room temperature) and 12 hours (refrigerated).
4. EDTA specimen for reticulocyte counts - stable for 6 hours (room
temperature) and 72 hours if refrigerated.
5. Glucose test drawn in sodium fluoride tubes - stable for 24 hours (room
temperature) but stable for up to 48 hours when refrigerated at 2°C to
8°C.
6. Prothrombin time (PT) - stable for 24 hours and partial thromboplastin
time (PTT) should analyzed within 4 hours from collection.
CENTRIFUGATION
✓ A centrifuge is a apparatus that is used to separate cells, plasma or
serum of blood specimen which is achieved by spinning the blood
tubes inside the vessel at a high speed such that the centrifugal force
will cause the separation of specimens.
✓ It is important to leave the stoppers on the tube before and during
centrifugation to avoid contamination, evaporation, aerosol
formation, and pH changes which will affect the accuracy of the
results.
✓ Take note that the tubes should be balanced in a centrifuge, meaning
tubes of the same size and volume of specimen should be placed
opposite one another. A centrifuge should not be repeated.
✓ The plasma specimen collected in tubes with anticoagulants should
be centrifuged immediately and without any delay. On the other
hand, serum specimen needs to be completely clotted prior to
centrifugation. Normally, complete clotting takes around 30 to 60
minutes at room temperature.
✓ Although there are machines that tests sample specimens directly
through the stopper, most of the test needs the stopper to be
removed to obtain the serum or plasma. Removal can be done using
removal devices or robotics but if there is a need to remove the
stopper manually, a face shield or splash shield should be used to
protect the technician from any risk of contamination. A gauze or
tissue is used to cover the stopper to catch drops of blood that may
leak or to catch aerosol that maybe released during the process.
✓ The tube stoppers should be removed by pulling it straight up and off
the tube.
ALIQUOT PREPARATION
✓ An aliquot of specimen refers to a portion of a sample specimen
taken for chemical analysis or testing.
✓ This is prepared when multiple tests are ordered on a single
specimen.
✓ The preparation is done by transferring a portion of the specimen
into one or more tubes that are labeled with the same information as
the original specimen tube using a disposable transfer pipettes.
✓ Do not put specimen with different anticoagulants in the same
aliquot tube.
✓ Make sure to cover the tube as soon as it is filled.
OSHA-REQUIRED PROTECTIVE EQUIPMENT WORN WHEN
PROCESSING SPECIMEN
When processing specimen in the laboratory, the health worker is
exposed to blood and other potentially infectious materials. For this
reason, health care institutions should comply with the appropriate
protective equipment required by the Occupational Safety and Health
Administration (OSHA) which includes wearing gloves to prevent
contact with blood, laboratory gown, laboratory coats, and
masks.