MLSPLM2C: PRINCIPLES OF MEDICAL LABORATORY SCIENCE 2 | LECTURE
CAPILLARY PUNCTURE
3RD SEMESTER | S.Y. 2022-2023
LECTURER: MS. MARY GRACE P. CARDENAS
CAPILLARY PUNCTURE
TYPE OF EQUIPMENT FOR CAPILLARY
COLLECTION
The following are the equipment used for capillary
collection:
• Lancet/Incision Devices
• Laser Lancet
• Micro collection Containers
• Microhematocrit Tubes and Sealants
• Microscope Slides
• Warming devices
• Capillary Blood Gas Equipment (CBG)
COMPOSITION OF CAPILLARY BLOOD
SPECIMEN, THE TEST AND REFERENCE
VALUE
1. The Capillary blood specimen
2. Arterial Blood
3. Venous Blood
4. Capillary Blood
INDICATIONSO F PERFORMING
CAPILLARY PUNCTURE ON ADULTS,
CHILDREN, AND INFANTS
1. Veins are fragile and not accessible
because of scars, burns, etc.;
2. Veins are reserved for another procedure
such as chemotherapy;
3. Clotting tendencies,
4. Extreme fear of needles and
5. Veins will be used for POCT such as
glucose monitoring.
This is also the preferred method for infants and
young children because of the following reasons:
1. Health risks such as anemia, and cardiac arrest;
2. Requires only a small volume of blood;
3. Venipuncture could damage veins and tissues
surrounding the site;
4. Puncturing could result to hemorrhage,
thrombosis, gangrene, and infection;
5. Risk of injury because of the restraint needed in
venipuncture; and
6. Capillary blood is the preferred specimen.
ORDER OF DRAW FOR COLLECTING
CAPILLARY SPECIMEN
Order of Draw refers to the specific sequence with
which blood must be drawn and collected in tubes
to avoid cross contamination of additives between
the tubes. The Order of Draw should be Blood gas
specimens (CBGs), EDTA specimens, other
additive specimen and serum specimen.
PREPARED BY: SANTOS, ALTHEA V.
ROUTINE AND THICK BLOOD SMEARS
• Routine blood film/smear preparation is a
blood test that is used to check abnormalities
in the blood cells.
• A small drop of blood is placed near the frosted
end of the glass side. Another slide is used to
spread the blood in a thin film over the slide. It
is then air-dried and stained.
• Thick blood smear preparation is used to
determine if the patient has malaria which is
diagnosed by its presence in the peripheral
blood smear.
COLLECTION OF SPECIMEN FOR
CAPILLARY GAS, NEONATAL BILIRUBIN,
AND NEWBORN SCREENING TESTS AND
ITS CLINICAL SIGNIFICANCE
• Capillary blood gas specimen by heel puncture
which is recommended for infants and small
children. The blood samples are collected on
the same site as routine capillary puncture
specimens.
• Neonatal bilirubin collection is used to help
determine any liver disorder in infants. This is
collected with a heel stick.
• Newborn/neonatal screening is done as part of
the routine check for infants to determine
inborn disorders.
• Newborn screening blood spot collection is
done 24 to 48 hours after the baby is born,
where a few drops of blood are collected
through heel stick to determine disorders that
are not apparent at birth that could lead to
disability or even death.
PERFORMING A CAPILLARY PUNCTURE
A. INDICATION
1. METHOD CHOICE FOR:
a. Children under 1 year of age (foot)
b. Adults with inaccessible of veins (finger)
2. USED FOR ADULTS PATIENTS:
a. Who are severely burned
b. With cancer and whose veins are reserved for
the therapeutic purposes
c. Who are obese and whose veins are too deep
to locate
d. Who are geriatric and whose veins are very
fragile
e. who performs test on themselves
• e.g. Point of care testing (POCT) for
glucose monitoring
3. OTHERS:
a. Procedure that requires capillary blood (e.g.,
malarial smears)
Page 1 of 2
 B. COLLECTION SITE
1. The last phalanx of the 3rd (great/middle) and
4th (ring) finger (note: cut across the finger
print line)
2. Outer area of the plantar surface of the foot
(heel stick)
3. Fleshy portion of the earlobe (e.g. burn patien)
C. MATERIALS
1. ALCOHOL
a. Clean finger with 70% isopropyl alcohol
b. Do not use povidone-iodine (↑ Potassium
,phosphorus, uric acid)
2. GAUZE AND BANDAGES
3. LANCETS
a. Retractable puncture device (e.g. guillotine-
type or half-moon type cut) with controlled
depth
i. The blade is pushed into the skin
ii. Pressed against the skin
b. The puncture of choice for infants is the heel
c. Optimal Depth without injury
i. 0.85 mm for premature infants
ii. 2 mm for full term infants
4. WARMING DEVICES
a. Enlarge capillaries
b. By wrapping a washcloth
i. Ideal temp. is 42oC
ii. Wrapped for 3-5 mins.
5. Micro collection Tubes
a. Do not scrape of the blood from the surface of
skin
b. Rotate the tube after every drop
c. Anticoagulated tube should be mixed 8-10
times
D. ORDER OF DRAW
1. Blood Gases (Heparin)
2. EDTA
3. Sodium Heparin
4. Other additive
5. Nonadditive
E. LABORATORY VALUES

SS

Page 2 of 2
 MLSPLM2C: PRINCIPLES OF MEDICAL LABORATORY SCIENCE 2 | LECTURE
SPECIAL COLLECTION PROCEDURES
3RD SEMESTER | S.Y. 2022-2023 PREPARED BY: SANTOS, ALTHEA V.
LECTURER: MS. MARY GRACE P. CARDENAS

PATIENT IDENTIFICATION, TYPES OF STERILE TECHNIQUE IN BLOOD

SPECIMEN, LABELING PROCEDURES CULTURE COLLECTION
REQUIRED FOR BLOOD BANK TESTS • A blood culture is a test that checks the blood
for pathogens for patients who have a fever of
unknown origin (FUO).
• The test determines the presence of bacteria in
the blood that causes bacteremia or presence
of microorganism and toxins in the blood that
causes septicemia.
• Aseptic Technique
• The physician orders this test only if there is a
The following information must be included: probability of bloodstream invasion.
1. Full name including middle initial • For optimum results, the collection should have
2. Hospital ID Number 2-4 blood culture sets placed in special bottles,
3. Social Security Number for outpatients one aerobic (with air) and one anaerobic
4. Date of Birth (without air), that were drawn 30 to 60 minutes
5. Date & time of collection apart.
6. Initials of the phlebotomist. • Skin antisepsis is crucial in the blood collection
Note: Room number and bed number is procedure because it prevents the
optional. contamination by microbial flora in the skin
• Blood banks can use special identification which can be introduced in blood culture
systems such as ID bracelet (self-carbon bottles and affect the results.
adhesive for specimen), Blood ID band (linear MEDIA INOCULATION METHODS
bar-code), and Siemens Patient Identification There are three ways to inoculate the medium:
check-blood administration. 1. Directly into the bottle (during collection),
• ü Screening includes typing the blood for 2. Collected in a syringe(after collection),and
transfusion and screening for infectious 3. Through an intermediate collection tube (in the
diseases. The blood type (ABO) and Rh factor laboratory).
(+ or -) are determined. • When using the syringe method, blood is transferred to the
• ü There are cases when there is a need toculture bottles after completing the draw using a safety
conduct a cross- match test to eliminate bloodtransfer device which is activated as soon as the needle is
related compatibility issues that may ariseremoved from the site.
between the patient and the donor. • Intermediate Collection Tube is performed in the laboratory
PROPER DONOR IDENTIFICATION, rather than the patient's bedside.
BLOOD UNITS LABELING AND POST COAGULATION SPECIMEN AND ITS
HANDLING PROCEDURES PROPER COLLECTION AND HANDLING
• Blood donor collection is done mainly for the • A physician requests coagulation tests to
purposes of blood transfusion. assess the blood clotting functions especially if
• Blood donors should be 17 to 66 years of age, the patient has an unexplained bleeding.
with a minimum weight of 110 lbs. They must • This test could be prothrombin time (PT),
have completed the physical exam and activated partial thromboplastin time (aPTT),
declared their medical history. and thrombin time (TT) which is a close
• The unit of blood can be separated into RBCs, approximation of the hemostatic system.
plasma and platelets but these should still be • The integrity of the blood specimen during
traceable to the donor. collection and transportation has a direct effect
• There are also cases when there is an on the test result, thus, special care should be
autologous donation where the patient donates taken to ensure that it is done properly.
blood for his use especially for elective • If only the coagulation specimen is drawn, draw
surgeries. a "clear" tube with 1-2 mL which is discarded
• Another special scenario is cell salvaging prior to collection. The blood specimen is then
where the blood of the patient is salvaged collected using a light blue top with 3.2% citrate
(during surgical procedure), washed and re- tube with the correct (9:1) blood to coagulant
infused after testing for residual free ratio.
hemoglobin.

Page 1 of 5
• To ensure proper mixing, invert the tubes
gently 3 to 4 times immediately after collection.
• Coagulation sample is collected second when
an evacuated tube system is used.
• For draws using indwelling catheter, the line
should be flushed with 5 mL of saline. The 5 mL
which is equivalent to six dead space volumes
of catheter is discarded.
• The concentration of the sodium citrate should
be adjusted for patients with above 55%
hematocrit values.
• The collected specimen should be transported
to the laboratory immediately. Should the
specimen be unable to reach the laboratory
within four hours, it should be centrifuged and
the plasma frozen.
2-hour Postprandrial Glucose
This blood test is done to check if the patient is
suffering from diabetes and other metabolic
problems. The 2-hour PP test is also used to
monitor insulin therapy. The principles of 2-
hour PP specimen collection are:
1. A high-carbohydrate diet was introduced 2 to 3
earlier.
2. The patient should fast at least 10 hours prior
to the test.
3. Fasting glucose specimen maybe be collected
before the start of the procedure.
4. A special breakfast containing an equivalent of
100g glucose or a glucose beverage will be
given on the day of the test.
5. Blood glucose specimen will then be collected
2 hours after the meal.
Glucose Tolerance Test
• A patient who could be suffering from
carbohydrate metabolism problems is
subjected to the glucose tolerance test (GTT)
which is also called oral glucose tolerance test
(OGTT) to evaluate the ability of the body to
metabolize glucose by measuring the
tolerance level to high glucose level.
• Insulin response to a measured dose of
glucose is recorded by specimen collection at
given intervals.
• The GTT length is 1 hour for gestational
diabetes while it is 3 hours for other
evaluations.
• The method of collection should also be
consistent be it venipuncture or skin puncture.
GTT Procedure
Before the procedure the patient must eat
balanced meal containing approximately 150
grams of carbohydrates for 3 days and must
fast for 12 to 16 hours before the scheduled
test. Drinking water is allowed to avoid
dehydration and because urine specimen is
also collected as part of the test. The steps in
the GTT procedure are as follows:
1. Begin with the normal identification protocol.
Explain the procedure and advise the patient
that only water is allowed during the whole test
period.
2. The fasting specimen is drawn and the glucose
level is checked and should be over 200 mg/dL
for the test to proceed.
3. The patient should collect a fasting urine
specimen if ordered.
4. The patient is given the glucose beverage
dose. Adult dose is 75g while children are given
1g per kilogram of weight. For gestational
diabetes the dose should be between 50 to
75g.
5. The beverage should be ingested within 5
minutes.
6. Record the time when the drink was finished
then start timing the test which is collected
within 30 minutes, 1 hour, 2 hours and so forth.
7. A copy of the collection time is provided to the
patient.
8. If applicable the collection time for other
specimen such as urine should coincide with
the computed collection time.
9. The exact time collected and the time interval
should be written in the label along with patient
identification information.
10. Transport the specimen immediately or within
2 hours for accurate results.
Lactose Tolerance Test
A lactose tolerance test measures the ability
of the body to process lactose and determines
if the patient lacks mucosal lactase which is an
enzyme that converts lactose into glucose or
galactose. Gastrointestinal distress and
diarrhea follows after ingestion of milk or food
containing lactose for those who lack the
lactase enzyme.
The principles for the lactose tolerance testing
include:
1. It is recommended that a 2-hour GTT test be
administered at least a day before the lactose
tolerance test.
2. Equal amount of lactose is substituted to
glucose but the test will be performed following
the manner of GTT.
3. Glucose specimen is drawn at the same time
as the previous GTT procedure.
4. The GTT curve and the glucose curve will be
similar if the patient has mucosal lactase.
5. If the patient is lactose intolerant, the result will
yield a "flat" curve where glucose levels just
slightly rise over the fasting level.
6. Patients with slow gastric emptying, Crohn's
disease and cystic fibrosis could show false-
positive results.
Page 2 of 5
 Paternity/Parentage Testing
Parentage testing or paternity testing is
performed to exclude the possibility of paternity
of a particular child.
Should the result does not exclude alleged
parent further test is performed which include
extended red cell antigens, red cell enzymes,
serum proteins testing, white cell enzymes,
white cells antigen, and human leukocyte
antigen (HLA).
DNA paternity testing is the most advanced
technology to date.
The test uses the genetic fingerprinting or DNA
profiling to establish parentage by providing
genetic proof.
Polymerase chain reaction (PCR) and
restriction fragment length polymorphism
(RFLP) are two other methods used today
although the older techniques still exist.
The principles of DNA paternity testing are:
1. All involved parties need to submit a
government issued photo identification along
with the completed Chain of Custody form.
The photos of all tested parties are also taken.
2. The Buccal samples as collected by using a
swab that was rubbed inside the cheek and the
loose cheek cells adhere to the swab.
3. Sealed and tamper-evident package are used
to hold the specimen during transportation to
the laboratory.
4. The test results are ready after 48 hours which
are usually sent via mail.
CHAIN OF CUSTODY PROCEDURES IN
DRUG TESTING
Toxicology Specimens
A phlebotomist plays a role in both clinical
toxicology (detection of toxins and treatment)
and forensic toxicology (legal consequences of
toxin exposure) because toxicology test
involves collection of blood, hair, urine, and
other substances from the body for the purpose
of determining the presence of toxins which
could be in very small amounts.
Forensics Specimens
Toxicology specimens are ordered by the law
enforcement department for legal or forensic
purposes.
The most common specimen are breath and
blood for alcohol.
Others request urine for drug screening, blood
specimen for drugs and DNA analysis.
For forensic specimen, there is a need to track the
specimen from the time of collection until the
time that the results are released using a
special protocol called chain of custody.
TIMING, ROLE OF DRUG HALF-LIFE,
NAMES OF DRUGS, ITS PEAK, TROUGH,
AND THERAPEUTIC LEVELS IN DRUG
MONITORING
Therapeutic Drug Monitoring
• The Therapeutic drug monitoring (TDM)
measures drug levels at designated intervals
so that the appropriate dosage can be
established and maintained for the patient thus
avoiding toxicity.
• TDM begins with prescription of the initial
dosage appropriate for the patient's clinical
condition. The amount in the bloodstream is
expected to rise, eventually reach peak
(maximum) which screens drug toxicity, and
gradually fall to a trough or minimum level
which ensures that the levels within therapeutic
range.
• The timing of collection is important for
aminoglycoside drugs (amikacin,
gentamicin, and tobramycin) which have short
half-lives but it is not critical for phenobarbital
and digoxin.
• Appropriate concentrations should be given to
optimize the clinical outcomes while
considering the drug dosage, history of dosage
given, the recorded response of the patient and
desired outcome.
Therapeutic Phlebotomy
• Therapeutic phlebotomy is performed by
drawing a large volume of blood from the
patient about 500 mL as part of the treatment
procedure for polycythemia and
hemochromatosis.
• Polycythemia is the overproduction of red
blood cells which is harmful to the patient. A
hematocrit test is used to check the RBC level
and removal of blood is done when level
exceeds a certain level with the goal of
returning the levels to normal range.
• Hemochromatosis is characterized by excess
iron deposits in the tissues which could be due
to problems with iron metabolism because of
multiple blood transfusions or excessive intake
of iron.
Blood Alcohol (Ethanol) Specimens
• Blood alcohol (ethanol [ETOH]) tests are
usually ordered for purposes related to
treatment but could also be for industrial or job-
related purposes such as insurance claims or
programs and employee drug screening.
• The law enforcement department orders blood
alcohol concentration (BAC) for individuals
involved in traffic related accidents.
• The ETOH test for treatment purposes do not
require the chain of custody to be
accomplished but the results of such tests can
become evidence in court. However, BAC for
Page 3 of 5
 industrial and legal samples should follow the
chain-of- custody protocol.
• The ETOH specimen collection uses aqueous
povidone-iodine and aqueous benzalkonium
chloride (BZK).
• Avoid using isopropyl alcohol and tincture of
iodine as antiseptic because these might affect
the results.
• Use gray-top sodium fluoride glass tubes
for specimen collection. These tubes could be
with anticoagulant, but it depends on the need
of the required specimen for a specific test.
• The tubes are filled until the vacuum is
exhausted. The tube stopper should always
remain in place because alcohol could
evaporate.
Drug Screening
• Companies, healthcare organizations and
sports associations subject their potential
employee to drug screening as part of their pre-
employment requirement.
• The company or organization could also run a
random screening without prior notice.
• The specimen used is urine instead of blood.
The chain of custody protocol is strictly
implemented since legal implications are
involved.
Patient preparation requirements
1. The purpose and procedure should be
explained to the patient.
2. The patient should be advised about his legal
rights.
3. There should be a witness present when the
form is signed.
Specimen collection requirements
1. A special area should be designated for the
purpose of urine collection.
2. During the collection, a proctor is present to
ensure that the specimen came from the
correct person.
3. Split sample may be used for parallel testing.
4. Proper labeling should be followed to establish
a chain of custody.
5. Protect the specimen from tampering. After
collection it should be sealed in a lock
container and sent to the laboratory
immediately. Documentation should be
handled carefully.
Trace Elements
• Trace elements tests for presence of
aluminum, arsenic, copper , lead, iron, and
zinc.
• They are collected in small amounts and must
use special element-free tubes colored royal
blue, since traces of these elements in the
glass, plastic or stopper could trickle into the
specimen will affect the accuracy of the result.
• The type of additive, if any, is indicated in the
label. (red - no additive, lavender - EDTA, and
green-heparin). To avoid contamination,
change the transfer device before filing the
tube.
Coagulation Monitoring by POCT
• Warfarin and heparin therapy can be
evaluated using a Coagulation POCT
analyzer.
• The common coagulation tests checked are
Prothrombin time (PT) & international
normalized ratio (INR), Activated partial
thromboplastin time (APTT or PTT), Activated
clotting time (ACT), and Platelet function.
• Listed are the available POCT instruments
available:
o Cascade POC —ACT, APTT, PT/INR
o CoaguChek XS Plus —PT/INR
o GEM Premier 4000 —ACT, APTT,
PT/INR
o i-STAT —ACT, PT/INR
o Verify Now —platelet function
Bleeding Time
• Bleeding time (BT) test is a non-
instrumented test ordered by the physician to
evaluate the capillaries for platelet plug
formation which is indicative of disorder in the
platelet function or problems in capillary
integrity.
• This test is also used for pre-surgical screening
and detection of problems involving
hemostasis. Although platelet function assays
(coagulation test) have been mostly ordered in
place of BT, it is still requested on some
occasion.
• Execution of the proper technique is important
to get an accurate result.
• The site should be in the volar lateral of the
forearm and 5 cm distal to the antecubital
area.
• The bleeding time is recorded and rounded to
the nearest 30 seconds. Normal results are
between 2 to 8 minutes.
Arterial Blood Gases
• The arterial blood gas (ABG) test measures
the level of oxygen, carbon dioxide and acid-
base (pH) in the patient's blood which gives the
physician an idea about the status of the
function of your lungs, heart and kidneys.
• ABGs measured by POCT methods include
potential hydrogen (pH), partial pressure of
carbon dioxide (PCO2), partial pressure of
oxygen (PO2), and oxygen saturation (SO2).
• pH refers to the concentration of hydrogen ions
in a solution. The arterial pH test checks the
balance of the acid- base level which shows
the metabolic and respiratory status of the
patient.
Page 4 of 5
• Normal range is from 7.35 to 7.45 only.
• The PCO2 is an indicator on how well air is
exchanged between the blood and lungs. The
test shows the measure of pressure exerted by
dissolved CO2 in the blood plasma in
proportion to the PO2 in the alveoli.
• Hypoventilation is when the PCO2 level
increased to an abnormal level while
hyperventilation is when it decreases.
• PO2 is representative of the pressure exerted
by the dissolved O2 and the ability of the lungs
to diffuse oxygen through the alveoli which is
usually used to measure the effectiveness of
an oxygen therapy. A normal person exhibits
98% oxygen saturation.
Electrolytes
• The electrolytes in the body mainly aid in
moving nutrients in the body and removes
wastes in the cells of the body.
• The POCT uses electrolyte panels to
determine the blood level of sodium (Na+),
potassium (K+, chloride (Cl- ), bicarbonate
ion (HCO3), and ionized calcium (iCa2+).
• Sodium helps keep the normal balance of fluids
in the body and plays a role in transmitting
nerve impulses. An elevated level of sodium is
called Hypernatremia while reduced level is
known as hyponatremia.

Page 5 of 5
 MLSPLM2C: PRINCIPLES OF MEDICAL LABORATORY SCIENCE 2 | LECTURE
HANDLING AND PROCESSING OF BLOOD & NON-BLOOD SPECIMENS FOR LABORATORY TESTING
3RD SEMESTER | S.Y. 2022-2023
LECTURER: MS. MARY GRACE P. CARDENAS
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 phase mishandling or error.
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 - usually between 3
to 10 inversions.
• Transporting specimens - 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 that precipitate or agglutinate if
allowed to cool below body temperature
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, cyrofibrinogen, and
cyroglobulins
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
PREPARED BY: SANTOS, ALTHEA V.
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 reasons.
The specimen is rejected if it did not meet a
specific criterion such as:
• Specimen is not identified properly.
• It has inadequate volume to complete the test.
• Hemolysis
• The use of wrong tube for collection.
• Outdated tube
• Improper handling (improper mixing)
• Contaminated specimen
• Insufficient specimen or "Quality Not Sufficient"
(QNS)
• Incorrect collection time
• Exposure to light
• Did not follow testing time limits
• 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
Page 1 of 6
and testing. Other exceptions to the time limit rule
are as follows:
1. Blood smear from EDTA specimen
2. EDTA specimen for CBC
3. EDTA specimen for erythrocyte
sedimentation rate (ESR) determination
4. EDTA specimen for reticulocyte counts
5. Glucose test drawn in sodium fluoride tubes
6. Prothrombin time (PT)
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.
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.
• Most of the test needs the stopper to be
removed to obtain the serum or plasma.
• 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.
THE DIFFERENT NON-BLOOD
SPECIMENS FOR LABORATORY TESTING
• A phlebotomist maybe involved in collection
and handling of specimen other than blood, or
non-blood specimen such as urine, amniotic
fluid, saliva, semen, sputum, etc.
• To avoid delays in testing, he should check and
verify information on the label of the specimen
before receiving them.
TESTS PERFORMED, COLLECTION AND
HANDLING PROCEDURES FOR THE
DIFFERENT NON-BLOOD SPECIMENS
The non-body fluid specimens are substances
produced by the body which can be in liquid or
semi liquid form. These are found within body
organs such as the bladder and spaces such as
the joints.
Urine
• The procedure for collection should be
explained properly to the patient since the type
of urine collection and the method of collection
depends on the type of test ordered by the
physician.
• ü The urine analysis also helps in (1)
monitoring wellness, (2) determination and
treatment of urinary tract infections, (3)
detection and monitoring progress of treatment
in metabolic diseases, and (4) determines the
effectiveness of an administered therapy as
well as checking if the patients exhibited any
therapy-related complications.
The accuracy of urine analysis results will largely
depend on several factors such as:
o the collection method,
o container use,
o transportation and handling of specimen
and
o he timeliness of the testing.
Common Urine Test
Routine urinalysis (UA)
• This frequently ordered urine test screens the
patient for any urinary or systematic disorders.
• This can be ordered during hospitalization and
also form part of the physical examination
procedure.
• The analysis is not limited to physical analysis
(color, clarity and odor) but also covers the
chemical (ph, specific gravity, detection of
bacteria, etc.) and microscopic analysis (urine
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 components such as cells, crystals and
microorganisms) of the specimen.
• Although random specimen is acceptable,
midstream collection is recommended and
ideal to ensure that there is no contamination
caused by genital secretions, pubic hair or
bacteria surrounding the urinary opening.
• The specimen container used for collection
should be clear and dry with tight-fitting lids and
they should be chemically cleaned.
• After collection, the specimen should be
transported to the laboratory immediately but it
could be held at room temperature for 2 hours
or it could be refrigerated if transport will take
longer than prescribed.
• Make sure that the specimen is protected from
light.
Culture & sensitivity (C&S)
• The Culture & sensitivity (C&S) urine test is
requested if the patient has symptoms of a
urinary tract infection (UTI).
• The urine specimen used should be midstream
clean-catch collection which is placed in a
sterile container.
• A measured portion of urine is cultured on a
special nutrient medium for 18 to 24 hours.
• If a microorganism is present and identified, a
sensitivity or antibiotic susceptibility test is
performed to determine the most effective
antibiotic that should be prescribed for
treatment.
Urine cytology studies
• The urine cytology study on urine is requested
by the physician to detect cancer,
cytomegalovirus, & other viral and
inflammatory diseases in the urinary system.
• A fresh clean-catch specimen is required.
• A smear from the cells of the lining of the
urinary tract is stained using the Papanicolaou
(PAP) method and examined under the
microscope for abnormal cells.
• The collected specimen should be examined
immediately but if delays could not be avoided,
the specimen is preserved by adding an equal
volume of 50% alcohol.
Urine drug testing
• Drug screening is performed using urine since
many drugs are detected in the urine but not in
the blood. This test is performed to detect:
1. illicit use of recreational drugs,
2. use of anabolic steroids to enhance
performance in sports, and
3. unwarranted use of prescription drugs. It
also monitors therapeutic drug use to
minimize the symptoms associated with the
withdrawal and also confirm drug overdose.
Urine glucose and ketone testing
• The urine glucose and ketone testing is
performed to screen diabetes and determine
the glucose level for patients who are already
diabetics.
• The urine ketone level test is used to determine
if the patient is suffering from diabetic
ketoacidosis.
• The color changes in the test strip are
compared to a color chart to interpret the result
of the test.
Urine pregnancy testing
• The urine pregnancy test is used to confirm
pregnancy which can be detected 8 to 10 days
from conception.
• The first morning urine specimen is preferred
because it has the highest concentration of
human gonadotropin (HCG).
Types of Urine Specimens
1. Random: collected at any time
2. First morning/8-hour urine specimen: collected
immediately upon waking up from an 8 hours
of sleep
3. Fasting: second specimen voided after fasting
(glucose monitoring)
4. Timed: collected at specific times or pooled
throughout a specific time period.
a. Tolerance test (glucose) - fasting, 1/2
hour, 1 hour, etc.
b. 2-hour postprandial - 2 hours after a
meal
c. 24-hour - collection & pooling of all
urine which is voided in 24 hours
d. Double-voided - wait for specific time
(30 minutes) after emptying the bladder
24-Hour Urine Collection Procedure
1. Empty the bladder upon waking up.
2. Affix the label in the container and write down
the time and date of the specimen collection.
Start timing.
3. Collect all the urine that has been passed for
the next 24 hours
4. If instructed, refrigerate the specimen collected
all throughout the collection period.
5. The urine is collected prior to bowel movement.
6. Drink fluid as needed to avoid dehydration.
7. Take one last void at the end of the 24 hour
collection period
8. Make sure to seal the container before placing
in the cooler. Arrange to transport the
specimen to the laboratory at the soonest
possible time.
Urine Collection Methods
1. Regular voided
2. Midstream
3. Midstream clean-catch
4. Catheterized
5. Suprapubic aspiration
6. Pediatric
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 Amniotic Fluid
• The amniotic fluid is a clear and slightly
yellowish liquid contained in the amniotic sac
that is surrounding and cushioning the fetus
during pregnancy.
• The specimen is collected by the physician
using transabdominal amniocentesis
preferably 15 weeks after gestation.
• The procedure is guided by the ultrasound
machine and about 10 mL of fluid is aspirated
from the amniotic sac thru the needle inserted
in the mother's abdominal wall into the uterus.
• The test is done to detect any genetic disorder,
check any problem in fetal development and
verify the gestational age.
Cerebrospinal Fluid
• The cerebrospinal fluid (CSF) refers the liquid
that surrounds brain & spinal cord.
• The testing is used to diagnose meningitis and
other disorders such as brain abscess, CNS
cancer and multiple sclerosis.
• The CSF is collected in 3 special sterile tubes:
first tube - chemistry and immunology test,
second tube - microbiology studies, and the
third tube for cell counts.
Gastric Fluid/Gastric Analysis
• The gastric analysis is performed to check the
contents of the stomach for abnormal
substances and evaluate the production of acid
by evaluating the gastric acid concentration.
• The sample is aspirated to determine acidity
before the stimulation. A gastric stimulant is
administered intravenously after the specimen
collection.
• A series of collection will follow with timed
intervals.
Nasopharyngeal Secretions
The Nasopharyngeal (NP) secretions specimen
are collected and cultured to determine the
presence of bacteria that causes:
1. diphtheria,
2. meningitis,
3. pertussis (whooping cough), and
4. pneumonia.
• Collection is performed by gently inserting a
dacron or sterile cotton-tipped flexible wire
swab into the nose into the nasopharynx (nasal
cavity & pharynx).
• The swab is rotated and gently removed then
placed in a properly labeled sterile container
containing transport medium for immediate
transport to the laboratory.
Saliva
• The saliva which is the fluid secreted by the
glands inside the mouth is used to check the
hormone levels and also determine alcohol
and drug or substance abuse.
• The collection of saliva is easy and non-
invasive because testing kits are available and
many are point-of-care test (POCT).
• The specimen needs to be frozen to ensure
stability prior to the delivery to the clinical
laboratory.
Semen
• The seminal fluid is a thick yellowish-white fluid
that contains sperm release during the male
ejaculation.
• Analysis is done to evaluate the fertility and
assess the effectiveness of sterilization after a
vasectomy procedure but it is also ordered for
forensic or legal reasons such as criminal
investigations involving sexual assault.
• The collected specimen is placed in sterile
container, kept warm and protected from light.
Serous Fluid
• The serous fluid is found between the
membrane that encloses that pleural,
pericardial, & peritoneal cavities.
• The fluid is collected by the physician by
aspiration and placed in tubes depending on
the type of test.
• EDTA tubes are used for cell counts and
smears, herapin or sodium fluoride tubes for
chemistry tests, non- anticoagulant tubes for
biochemical tests, and for cultures the
heparinized should be used.
Sputum
• Sputum (mucous or phlegm) specimen are
used to diagnose and/or monitor lower
respiratory tract infections like tuberculosis.
• It is recommended that the collection be done
first thing in the morning since larger volume
have accumulated overnight and at least 1
hour after meal to avoid gagging.
• The patient should gargle with water and
remove dentures (if any). The patient should:
1. take takes 3 or 4 deep breaths,
2. inhale and exhale,
3. cough forcefully on the last breath, and
4. expectorate and expel sputum into the
sterile container.
The procedure is repeated until adequate amount
is collected which is about 3 to 5 mL.
Sweat
• The sweat is used to analyze the chloride
content of patients under the age of 20 with
symptoms of cystic fibrosis (exocrine gland
disorder).
• During the sweat chlorides test, electrical
stimulation in the forearm or thigh
(iontoporesis) is used to transport the
pilocarpine (sweat-stimulating drug) into the
skin. The sweat is then collected, the weight is
recorded and the chloride content is analyzed.
• The sweat specimen can also be used to
determine illegal use of drug by placing
patches on the skin for an extended period of
time and which is further confirmed by
subsequent drug testing.
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 Synovial Fluid
The synovial fluid (viscous fluid that lubricates
movable joints) are tested to determine conditions
such as arthritis, gout and other inflammatory
conditions since increase in volume is attributed to
inflammation. These are collected in 3 tubes:
• EDTA/heparin: for cell counts, ID of
crystals, smear preparation
• Sterile: culture & sensitivity
• Non-additive: macroscopic appearance,
chemistry, immunology tests and observing
the clot formation
Buccal (Cheek) Swabs
• Buccal swabs are used to obtain loose cells
inside the cheek for purposes of DNA analysis.
• This is less invasive and painless alternative to
blood collection.
• The phlebotomist collects sample by placing
the swab inside the cheek and gently
massages the area.
• After the collection the swab is sent to the
laboratory where the DNA is extracted from the
cells in the swab.
Bone Marrow
• The bone marrow is examined to identify blood
diseases.
• The physician inserts a large-gauge needle
into the sternum (breastbone) or iliac crest (hip
bone) and aspirates 1.0 to 1.5 mL of specimen.
• A hematology technologist makes special
slides from the first marrow aspiration.
Breath Samples
• There are two types of testing for breath: C-
urea breath test (C-UBT) and Hydrogen breath
test.
• After the collection of the baseline breath
sample, the patient is asked to drink a special
substance that has synthetic urea.
• He is then asked to breath into a Mylar balloon
at specified intervals. The breath specimens
are analyzed for carbon-13 which confirms the
presence of the H. pylori.
• The Hydrogen breath test, on the other hand,
helps in the detection of carbohydrate
digestion problems such as lactose (milk
sugar) and fructose (fruit sugar).
• This test can also detect bacterial overgrowth
in the small intestine and is thought as the most
accurate tolerance test.
• In preparation for the test, the patient must not
take antibiotics 2 weeks prior to the scheduled
test.
• He should not eat certain food for 24 hours
before the test and should be on fasting on the
day of the test. Smoking and exercise is also
restricted 30 minutes before the test.
• The baseline breath sample is taken by
breathing into a special bag before the
introduction of a special drink.
• Additional breath samples are taken every 30
minutes for 3 hours and all are sent to the
laboratory for analysis.
Feces (Stool)
The fecal specimen (feces or stool) is collected to
1. determine gastrointestinal disorder,
2. analyze for presence of intestinal ova &
parasites (O&P),
3. cultured for the presence of pathogenic
bacteria and viruses,
4. check fat and urobilinogen content,
5. test for presence of occult blood. They are
collected in a clean and wide-mouth container
that are sealed and sent to the laboratory
after collection.
Hair
Aside from using the hair sample for trace & heavy
metal analysis, it is also used to detect chronic
drug abuse where hair is the preferred specimen
because not only is it easy to obtain, but it is also
not easy to alter or tamper.
Throat swabs
• Throat swabs are mostly collected to aid in
streptococcal (strep) infection detection.
• A special kit contains a sterile polyester-tipped
swab and a covered transport tube.
• The tube contains transport medium.
The throat culture specimen collection procedures
are as follows:
1. Follow hand hygiene and PPE procedures
2. Open the container to remove the swab in
an aseptic manner.
3. Stand to the side of the patient or stand back
to avoid droplet contact when the patient
coughs.
4. Ask the patient to tilt his head back and
open his mouth wide.
5. Examine the throat by directing a flashlight
to the back of the throat.
6. Use a tongue depressor to depress the
tongue and ask the patient to say "Ah" to
raise the uvula and get it out of the way.
7. Swab the tonsils, tonsilar crypts, back of the
throat and any area that has ulceration and
inflammation but avoid touching the lips,
tongue and uvula.
8. Discard the tongue depressor only after the
swab has been taken out of the mouth.
9. The swab is placed in the transport tube
(dipped in the medium) and the covered
carefully.
10. Follow proper labeling procedure for the
specimen.
11. Remove the gloves properly & apply proper
hand decontamination technique.
12. The collection tube is sent to the laboratory
for immediate processing.
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 Tissue Specimen
• The tissue specimen is usually collected using
biopsy where the tissue sample is removed for
examination.
• The phlebotomist should check the proper
handling procedure if the specimen arrived not
immersed in solution.
• In the case of genetic analysis, the tissue
samples are not to be placed in formalin.
• Improper handling is costly, inconvenient and
cannot be easily repeated.

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