ANALYSIS OF URINE AND OTHER BODY FLUIDS

THIRD YEAR | FIRST SEM | FINALS

SEMEN SPECIMEN COLLECTION
• Advances in the field of andrology and assisted • Abstinence of 2-3 days but not more than 7 days.
reproductive technology (ART), and increased concern • Collect the entire ejaculate. When a part of the first
over fertility, particularly by couples choosing to have portion of the ejaculate is missing, the sperm count will
children later in life, have resulted in increased be decreased, the pH falsely increased, and the
emphasis on semen analysis. specimen will not liquefy. When part of the last portion
• Patients with abnormal results on the routine semen of ejaculate is missing, the semen volume is decreased,
analysis performed in the clinical laboratory are often the sperm count is falsely increased, the pH is falsely
referred to as specialized andrology laboratories for decreased, and the specimen will not clot
further testing to determine the need for in vitro • Method of collection:
• fertilization (IVF). o Masturbation
• Clinical laboratory personnel may also be employed in o Coitus interruptus
andrology laboratories and perform routine and o Condom method (use only non-lubricant containing
specialized testing. rubber or silastic method)
• In addition to fertility testing, the clinical laboratory • Specimen should be delivered to the laboratory within 1
performs post-vasectomy semen analysis and forensic hour of collection (at room temperature)
analyses to determine the presence • Take note of the time of specimen collection, specimen
• of semen. receipt, and liquefaction.
• Analysis should be done after liquefaction 30-60 mins.
Definition of Terms If after 2 hours the specimen has not liquified, an equal
• Andrology - The study of diseases of the male volume of physiologic Dulbecco’s phosphate-buffered
reproductive organs saline or proteolytic enzymes such as alpha-
• Bulbourethral gland - Two small glands located on each chymotrypsin or bromelain may be added to induce
side of the prostate gland liquefaction and allow the rest of the analysis to be
• Epididymis - Small structure that forms the first part of performed.
the secretory duct of the testes erythrophagocytosis • Specimen awaiting analysis should be kept at 30
Engulf degrees Celsius.
• Liquefaction - The conversion of solid or coagulated
material to a liquid form MACROSCOPIC EXAMINATION
APPEARANCE Normal: Gray-white, translucent with
PHYSIOLOGY musty odor Increased white turbidity:
presence pf white blood cells (WBCs)
• Semen is composed of four fractions that are
and infection.
contributed by the testes, epididymis, seminal vesicles,
Red or brown coloration: the presence
prostate gland, and bulbourethral glands. of red blood cells
• Seminiferous tubules secrete sperm. Germ cells or the Yellow coloration: urine contamination,
production of spermatozoa are in the epithelial cells of specimen collection following
the seminiferous tubules. prolonged abstinence, and medications
• Sertoli cells provide support and nutrients for the germ VOLUME Normal: 2 to 5 mL
cells as they undergo mitosis and meiosis Increased in prolonged abstinence
(spermatogenesis). In the epididymis, the sperm mature Decreased in infertility, incomplete
and develop flagella. The entire process takes collection
approximately 90 days. VISCOSITY Normal: Discrete droplets
Abnormal: Threads (>2 cm long)
• Reasons for seminal fluid analysis includes the
↑ Viscosity: ↓ sperm motility
following:
Reporting:
✓ Fertility testing 0= watery
✓ post-vasectomy semen analysis 4= gel-like
✓ Forensic analysis May also be reported as low, normal or
high
pH Normal: 7.2-8.0
COMPOSITION OF SEMEN
↑pH: Infection
5% Spermatozoa 1. Seminiferous tubules ↓pH: ↑ Prostatic fluid
✓Spermatogenesis
✓Sertoli cells - serve as nurse SPERM CONCENTRATION
cells for developing sperm. • Normal value= 20 to 250 million sperm per milliliter
2. Epididymis
• The sperm are counted in the same manner as cells in
- sperm maturation (sperm become
motile) the cerebrospinal fluid cell count, that is, by diluting the
60-70% Seminal Seminal Vesicles specimen and counting the cells in the Neubauer
fluid provide nutrients for sperm and fluid chamber.
- rich in fructose for sperm motility • The most commonly used dilution is 1:20 prepared
20-30% Prostate Acidic fluid that contains ACP, zinc, using a mechanical (positive-displacement) pipette.
fluid citric acid, and other Dilution of the semen is essential because it immobilizes
enzymes for coagulation and the sperm before counting.
liquefaction • Diluents: formalin, sodium bicarbonate, saline, distilled
5% Bulbourethral Secretes thick alkaline mucus that water, tap water (cold)
gland neutralizes acidity from the • The back sides of the hemocytometer are loaded and
prostatic secretions and vagina.
allowed to settle for 3 to 5 minutes; then they are
counted, and the counts should agree within 10%.
• Makler counting chamber is used for undiluted
specimens and uses heat to immobilize sperms
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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
Calculating Sperm Concentration and Sperm Count: Additional Tests
• Short Method • Seminal fluid fructose
2 WBC squares= # sperms counted x 100,000 ✓ tested within 2 hours or frozen to prevent fructolysis
5 RBC squares= # sperms counted x 1,000,000 ✓ screening test is resorcinol test= (+) orange-red color
• Long Method
• Anti-sperm Antibodies
Sperm concentration (per mL) = # sperm counted x
dilution/area x depth (0.1) ✓ detected in semen, cervical mucosa, or serum
• Sperm count normal value is ≥ 40 million per ejaculate ✓ mixed agglutination (semen sample+ AHG+ latex
• Formula for sperm count: particles or treated RBC coated with IgG) - detects the
Sperm count = sperm concentration x sperm presence of IgG antibodies; normal = <10% motile
volume sperm attached to the particles
✓ Immunobead test - detects the presence of IgG, IgM,
SPERM MOTILITY and IgA antibodies; demonstrates what area of sperm
• Should be assessed using a well-mixed, liquefied semen (head, neck, tail) the autoantibodies affect.
specimen within 1 hour of specimen collection. • Microbial testing
• Sperm motility can be evaluated at room temperature or ✓ test for C. trachomatis, U. urealyticum, and M.hominis
37°C.
• When assessing motility at 37°C, the sample should be
incubated at this temperature and the preparation
made with prewarmed slides and cover slips.
• Read in 20 HPF

SPERM MOTILITY GRADING

GRADE WHO CRITERIA
4.0 a Rapid, straight-line motility
3.0 b Slower speed some lateral movement
2.0 c Slow forward progression, noticeable
lateral movement
1.0 d No forward progression
0 e No movement

• Computer Assisted Semen Analysis (CASA) provides

objective determination of both sperm velocity and
trajectory (direction of motion).
• Sperm concentration and morphology are also included
in the analysis

SPERM MORPHOLOGY
• Evaluated with respect to the structure of the head,
neckpiece, midpiece, and tail.
• Normal value:
✓ Routine criteria= 30% normal forms
✓ Kruger’s strict criteria= 14 % normal forms
• The normal sperm has an oval-shaped head
approximately 5 µm long and 3 µm wide and a long,
flagellar tail approximately 45 µm long.
• The acrosomal cap should encompass approximately
half of the head and cover approximately two-thirds of
the sperm nucleus.
• The midpiece is approximately 7.0 µm long and is the
thickest part of the tail because it is surrounded by a
mitochondrial sheath.
• Stains for sperm morphology:
✓ Wright’s,
✓ Giemsa,
✓ Shorr, or
✓Papanicolaou stain.
Note: Varicocele is the hardening of veins that drain the
testes. it is the most common cause of male infertility. sperm
head: tapered head.

SPERM VIABILITY
• Modified Bloom’s test
• Reagents = eosin and nigrossin
• Living sperms = unstained, bluish-white
• Dead sperm = red
• Normal value = 50% living sperms

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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
THE PROPER WAY TO PERFORM THE COLLECTION, SEMEN ANALYSIS The reference value for WHO
STORAGE, TRANSPORT AND ANALYSIS OF SEMINAL FLUID Criteria for Infertile Men
INTRODUCTION Volume < 2.0 mL
• Semen also known as seminal fluid pH < 7.2
• Consist of several fluids produced in various parts of the Sperm concentration <20 million sperm per mL
male reproductive system Total sperm number <40 million sperm per ejaculate
• Semen analysis is a vital step in the investigation of Motility <50% with progressive motility
several disorders affecting the male genital tract. Vitality <75% live
White blood cells >1 million per ml
• The study of semen parameters provides important
clinical information on the spermatogenesis and
NORMAL VALUES OF SEMEN ANALYSIS
functional competence of spermatozoa
PARAMETERS NORMAL VALUES
PHYSICAL EXAMINATION
PURPOSE
Appearance Gray-white to light yellow
• Fertility testing Volume 1.5-5 mL
• Post-vasectomy semen analysis Viscosity Pours in droplets
• Forensic analysis to determine the presence of semen Liquefies within 30 – 60
minutes after collection
COLLECTION, TRANSPORT AND STORAGE CHEMICAL EXAMINATION
• As per WHO recommendation – 2 semen samples pH 7.2-8.0
(interval should not be less than 7 days or more than 3 MICROSCOPIC EXAMINATION
months apart. Motility 40% or more with moderate to
• MTs should have clear instructions rapid forward progression
• Samples should be collected after a minimum of 2 days Sperm count 15 – 250 x 106 sperm/mL
(concentration)
(48 hours) and no longer than 7 days of sexual
Morphology 4% or more normal morphology
abstinence
Vitality 58% or more alive
• The time interval of the last ejaculation and the time of Leukocytes
the sample arrived in the laboratory must be indicated
in the request form
• A clean, wide-mouthed glass or plastic container
• Incomplete semen samples should be rejected
• Safety first

COLLECTION (Incorrect)
PART OF THE FIRST PORTION MISSING
✓ Decreased sperm count
✓ Falsely increased pH
✓ Specimen will not liquefy SPERM COUNT

PART OF THE LAST PORTION MISSING

✓ Decreased volume
✓ Falsely increased count
✓ Falsely decreased pH
✓ Specimen will not clot
EXAMINATION OF SEMEN
MACROSCOPIC EXAMINATION
• Volume
• Liquefaction time MORPHOLOGY
• Appearance
• ph
• Viscosity

MICROSCOPIC EXAMINATION
• Sperm count
• Sperm motility
• Sperm viability
• Sperm morphology
• Agglutination
• Antibodies coating of sperm

BIOCHEMICAL TESTS
• Fructose test
• Acid phosphatase

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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
SPERM/SEMEN ANALYSIS – ABNORMAL RESULTS
ABNORMALITIES DEFINITION
Aspermia Absence of semen
Azoospermia Absence of sperm in the semen
Hypospermia Low semen volume
Hyperspermia High semen volume
Oligozoospermia Very low sperm count
Polyzoospermia Abnormally high sperm count in
the ejaculate
Asthenozoospermia Poor sperm motility
Teratozoospermia Sperms that have morphological
defects
Necrozoospermia All the sperm in the ejaculate are
dead
Leucospermia A high level of white blood cells
present in the semen
Hematospermia Presence of RBC in the ejaculate

SEMEN ANALYSIS REPORT

Duration of abstinence: 6 days
Liquefaction time (min): 30 min
Interval between the start of ejaculation and analysis
(before liquefaction): 10 min
GROSS
Appearance: Creamy white
Consistency: Thin
Volume: 3.0 ml; pH: 8.0
Fructose: Present
Sperm Motility
a. Rapid progression: 5%
b. Slow progression: 25%
c. Non-progression motility: 5%
d. Immotile: 65%
Agglutination
Vitality
Count/ml: 5 million
MORPHOLOGY
Normal: 30%
Abnormal: 70%
Head defects: 25%
Tail defects: 30%
neck and mid-piece defects: 5%
Cytoplasmic droplets: 8%
headless ‘pinhead’: 2%
Pus cells: 1-2
RBC: 0-1
Epithelial cells: NI
Miscellaneous: NI

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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
FECAL ANALYSIS • Electrolytes - increased in secretory diarrhea and
PHYSIOLOGY negligible in osmotic diarrhea.
Normal fecal specimen • Fecal fluid pH (less than 5.6) - malabsorption of sugars
• Bacteria, cellulose, undigested foodstuffs, GI secretions, (osmotic diarrhea)
bile pigments, cells from intestinal walls, electrolytes, TECHNICAL TIP: Process specimens for osmolality testing
and water. immediately. Specimens that are stored for hours may have
• 100 – 200 g of feces (24-hour period) a markedly increased osmolality due to the increased
• Flatus – bacterial metabolism produces a strong odor degradation of carbohydrates
associated with feces and intestinal gas.
• Excessive gas production also occurs in lactose- SECRETORY DIARRHEA - caused by increased secretion of
intolerant people. water.
• Digestion of ingested proteins, carbohydrates, and fats • Bacterial, viral, and protozoan infections produce
takes place throughout the alimentary tract increased secretion of water and electrolytes
• Small intestine - is the primary site for final breakdown • Enterotoxin-producing organisms
and reabsorption. o Escherichia coli, Clostridium, Vibrio cholerae,
• Digestive enzymes secreted into the small intestine by Salmonella, Shigella, Staphylococcus,
the pancreas include; Campylobacter, protozoa, and parasites such as
○ TRYPSIN, CHYMOTRYPSIN, AMINO PEPTIDASE, AND Cryptosporidium
LIPASE OTHER CAUSES:
• Bile salts provided by the liver aid in the digestion of fats. • Drugs
• A deficiency in any of these substances creates an • Stimulant laxatives
inability to digest and, therefore, to reabsorb certain • Hormones
foods. • Inflammatory bowel disease (Crohn's disease,
• Excess undigested or unabsorbed materials then ulcerative colitis, lymphocytic colitis, diverticulitis)
appear in the feces, and the patient exhibits symptoms • Endocrine disorders (hyperthyroidism,
of maldigestion and malabsorption. • Zollinger-Ellison syndrome, VIPoma)
Under normal conditions; • Neoplasms
o Only between 500 to 1500 mL of this fluid reaches the • Collagen vascular disease
large intestine
o Only about 150 mL is excreted in the feces OSMOTIC DIARRHEA - caused by poor absorption that
o Large intestine is capable of absorbing approximately exerts osmotic pressure across the intestinal mucosa.
3000 mL of water • Incomplete breakdown or reabsorption of food
• Diarrhea- produced when the amount of water reaching presents increased fecal material to the large intestine,
the large intestine exceeds this amount, it is excreted resulting in water and electrolyte retention in the large
with the solid fecal material. intestine.
• Constipation - provides time for additional water to be • Maldigestion and malabsorption - contribute to osmotic
• reabsorbed from the fecal material, producing small, diarrhea
hard stools • The presence of unabsorbable solute increases the
stool osmolality and the concentration of electrolytes is
DIARRHEA AND STEATORRHEA lower, resulting in an increased osmotic gap.
• DIARRHEA - defined as an increase in daily stool weight • Causes of osmotic diarrhea
above 200 g, increased liquidity of stools, and o disaccharidase deficiency, malabsorption, poorly
frequency of more than three times per day. absorbed sugars, laxatives, magnesium-containing
• Classified in 4 factors; antacids, amebiasis, and antibiotic administration
o Illness duration
o Mechanism FECAL TESTS FOR DIARRHEA
o Severity, and • Laboratory testing of feces is frequently performed to aid
o Stool characteristics in determining the cause of diarrhea
• Acute diarrhea - less than 4 weeks
• Chronic diarrhea - more than 4 weeks

MAJOR MECHANISMS LABORATORY TEST USED

Secretory Fecal electrolytes (fecal
sodium, fecal
potassium)
Osmotic Fecal osmolality
Intestinal hypermotility Stool pH

• Normal total fecal osmolarity (290 mOsm/kg)

• Normal fecal sodium (30 mmol/L)
• Fecal potassium (75 mmol/L)
• Fecal osmotic gap
✓ Osmotic gap = 290-[2 (fecal sodium + fecal
potassium)]
• The osmotic gap in all forms of osmotic diarrhea is
greater than 50 mOsm/kg and less than 50 mOsm/kg
in secretory diarrhea.

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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
Differentiates the features of osmotic diarrhea and • Patients should understand that the specimen must not
secretory diarrhea. be contaminated with urine or toilet water
• Containers that contain preservatives for ova and
parasites must not be used to collect specimens for
other tests
• Random specimens suitable for qualitative testing for
blood and microscopic examination for leukocytes,
muscle fibers, and fecal fats are usually collected in
plastic or glass containers with screw-tops.
• For quantitative testing, such as for fecal fats, timed
specimens are required.
• Care must be taken when opening any fecal specimen
to slowly release gas that has accumulated within the
container.
ALTERED MOTILITY • Patients must be cautioned not to contaminate the
• Describes conditions of enhanced motility or slow outside of the container.
motility.
• Can be seen in irritable bowel syndrome (IBS) MACROSCOPIC SCREENING
○ extra sensitive, causing cramping, bloating, flatus, • The first
diarrhea, and constipation indication of GI
• IBS can be triggered by food, chemicals, emotional disturbances
stress, and exercise. can often be
• Intestinal hypermotility is the excessive movement of changes in the
intestinal contents through the GI tract. brown color
• Rapid gastric emptying (RGE) - describes hypermotility of and formed
the stomach and the shortened gastric emptying half- consistency of
time, which causes the small intestine to fill too quickly the normal
with undigested food from the stomach. stool.
• It is the hallmark of early dumping syndrome (EDS). • The
• Gastric emptying half-time range of 35 to 100 minutes appearance of
(varies age and gender) abnormal fecal
• Normal gastric emptying is controlled by fundic tone, color may also be caused by ingestion of highly
duodenal feedback, and GI hormones. pigmented foods and medications, so a differentiation
• RGE divided into early dumping and late dumping must be made between this and a possible pathologic
cause.
EARLY DUMPING LATE DUMPING o Color
SYNDROME SYNDROME o Appearance
symptoms begin 10 to 30 Occurs 2 to 3 hours
minutes following meal after a meal MICROSCOPIC EXAMINATION OF FECES
ingestion • It is performed to detect the presence of leukocytes
Nausea, vomiting, Characterized by associated with microbial diarrhea and undigested
bloating, cramping, weakness, muscle fibers and fats associated with steatorrhea.
diarrhea, dizziness, and sweating, and dizziness • FECAL LEUKOCYTES - seen in the feces in conditions
fatigue that affect the intestinal mucosa, such as ulcerative
colitis and bacterial dysentery.
STEATORRHEA (fecal fat) - useful in diagnosing pancreatic o Microscopic screening is performed as a
insufficiency and small-bowel disorders that cause preliminary test
malabsorption. o Presence or absence of fecal neutrophils can
• Absence of bile salts that assist pancreatic lipase in the provide the physician with diagnostic information
breakdown and subsequent reabsorption of dietary fat before receiving the culture report
(primarily triglycerides) produces an increase in stool fat • Specimens can be examined: as wet preparations
(steatorrhea) that exceeds 6 g per day. stained with methylene blue or as dried smears stained
• Steatorrhea may be present in both maldigestion and with Wright’s or Gram stain
malabsorption conditions and can be distinguished by
the D-xylose test.
• D-xylose is a sugar that does not need to be digested but
does need to be absorbed to be present in the urine.
• If urine D-xylose is low, the resulting steatorrhea
indicates a malabsorption condition.
• Malabsorption causes include bacterial overgrowth,
intestinal resection, celiac disease, tropical sprue,
lymphoma, Whipple disease, Giardia lamblia infestation,
Crohn's disease, and intestinal ischemia. • A lactoferrin latex agglutination test is available for
• Normal D-xylose test indicates pancreatitis detecting fecal leukocytes and remains sensitive in
refrigerated and frozen specimens.
SPECIMEN COLLECTION • MUSCLE FIBERS - Microscopic examination of the feces
• Stool specimen - a collection of a fecal specimen for undigested striated muscle fibers can be helpful in
• Patients should be instructed to collect the specimen in diagnosing and monitoring patients with pancreatic
a clean container. (bedpan or disposable container) insufficiency.
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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
• Increased amounts of striated fibers may also be seen • Cholesterol is stained by Sudan III after heating and as
in biliary obstruction and gastrocolic fistulas. the specimen cools forms crystals that can be identified
microscopically.

CHEMICAL TESTING OF FECES

Occult Blood
• Guaiac-Based Fecal Occult Blood Tests
• Immunochemical Fecal Occult Blood Test
• Porphyrin-Based Fecal Occult Blood Test

Occult Blood/fecal occult blood testing (FOBT)

• Undigested fibers have visible striations running both • most frequently performed fecal analysis
vertically and horizontally. Partially digested fibers • Methods for detecting fecal occult blood include the
exhibit striations in only one direction, and digested guaiac immunochemical, and fluorometric porphyrin
fibers have no visible striations. Only undigested fibers quantification tests. Immunochemical tests and fecal
are counted, and the presence of more than 10 is porphyrin quantification tests are more sensitive and
reported as increased specific methods than the guaiac-based fecal occult
• Patients should be instructed to include red meat in blood tests
their diet before collecting the specimen
• Specimens should be examined within 24 hours of Guaiac-Based Fecal Occult Blood Tests (gFOBT)
collection • most frequently used screening test for fecal blood ○
• QUALITATIVE FECAL FATS - Specimens from suspected • the least sensitive reagent, guaiac, is preferred for
cases of steatorrhea can be screened microscopically routine testing.
for the presence of excess fecal fat (steatorrhea). • commercial testing kits contain guaiac-impregnated
• The procedure can also be used to monitor patients filter paper enclosed in a cardboard slide
undergoing treatment for malabsorption disorders.
• In general, the correlation between the qualitative and
quantitative fecal fat procedures is good; however,
additional unstained phospholipids and cholesterol
esters are measured by the quantitative procedure.
• Lipids included in the microscopic examination of feces Immunochemical Fecal Occult Blood Test (iFOBT)
are neutral fats (triglycerides), fatty acid salts (soaps), • specific for the globin portion of human hemoglobin
fatty acids, and cholesterol. • uses polyclonal anti-human hemoglobin antibodies
• Their presence can be observed microscopically by • more sensitive to lower GI bleeding
staining with the dyes Sudan III, Sudan IV, or oil red O; • it doesn't require any dietary restrictions before the
Sudan III is the most routinely used. sample and testing can often be performed on a random
• The staining procedure consists of two parts: the neutral stool sample.
fat stain and the split fat stain.
• Neutral fats are readily stained by Sudan III and appear Porphyrin-Based Fecal Occult Blood Test
as large orange-red droplets. • The HemoQuant test includes the measurement of total
• Split fat stain representing total fat content can provide fecal hemoglobin or porphyrin derived from heme, an
a better indication intestinal converted fraction (ICF)
• more sensitive to upper GI bleeding
• not affected by the presence of reducing or oxidizing
substances or the water content of the fecal specimen

Quantitative Fecal Fat Testing

• used as a
confirmatory test
for steatorrhea
• requires the
• Soaps and fatty acids do not stain directly with Sudan III, collection of at
so a second slide must be examined after the specimen least a 3-day
has been mixed with acetic acid and heated. specimen
• Examining this slide reveals stained droplets that • The specimen is
represent not only the free fatty acids but also the fatty collected in a
acids produced by hydrolysis of the soaps and the large, pre-
neutral fats weighed
• Normal specimens may contain as many as 100 small container
droplets, less than 4 um in diameter, per high-power
field. The same number of droplets measuring 1 to 8 µm
is considered slightly increased, and 100 droplets Fat Measurement
measuring 6 to 75 µm is increased and commonly seen • Van de Kamer titration - routinely used, time-consuming
in steatorrhea. 12 An increased amount of total fat on • Gravimetric - measures all fecal fat
the second slide with normal fat content on the first slide
• Near-infrared reflectance spectroscopy – rapid
is an indication of malabsorption, whereas maldigestion
procedure for fecal fat that requires less stool handling
is indicated by increased neutral fat on the first slide.
by laboratory personnel
• Hydrogen Nuclear magnetic resonance spectroscopy
method-homogenized specimen is microwaved-dried
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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
and analyzed. The results correlate well with the
gravimetric method.

APT Test (Fetal Hemoglobin)

differentiate fetal blood from swallowed maternal blood in
the evaluation of bloody stools.

APT Test Procedure

1. Emulsify the specimen in water.
2. Centrifuge.
3. Divide the pink supernatant into two tubes.
4. Add 1% sodium hydroxide to one tube.
5. Wait 2 minutes.
6. Compare the color with that in the control tube.
7. Prepare controls using cord blood and adult blood.

Fecal Enzymes
Enzymes supplied to the gastrointestinal tract by the
pancreas are essential for digesting dietary proteins,
carbohydrates, and fats.

Pancreatic insufficiency
disorders such as chronic pancreatitis and cystic fibrosis.
Steatorrhea occurs, and undigested food appears in the
feces

Proteolytic enzymes
• Trypsin
• Chymotrypsin - more resistant to intestinal degradation,
capable of gelatin hydrolysis
• Elastase I - an isoenzyme of the enzyme elastase and is
the enzyme form produced by the pancreas

Carbohydrates
• The presence of increased carbohydrates in the stool
produces osmotic diarrhea
• May be present as a result of intestinal inability to
reabsorb carbohydrates, as seen in celiac disease, or;
• Lack of digestive enzymes such as lactase (lactose
intolerance), and Idiopathic lactase deficiency
• Most valuable in assessing cases of infant diarrhea
and may be accompanied by a pH determination
• Normal stool pH is between 7 and 8
• Copper reduction test is performed using a Clinitest
table

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ANALYSIS OF URINE AND OTHER BODY FLUIDS
THIRD YEAR | FIRST SEM | FINALS
THE PROPER WAY TO PERFORM FECAL OCCULT BLOOD OTHER NAMES
TEST (FOBT) • FOBT
INTRODUCTION • Stool occult blood
• Healthy adult – 2ml of blood per 150 grams of stool from • Occult blood test
the gastrointestinal tract daily • Hemoccult test
• More than the normal is considered significant, • Hidden blood test
however, no visible signs of bleeding may be present • Guaiac smear test
with this amount of blood • gFOBT
• Excessive bleeding may result – black, tarry stools and • Immunochemical FOBT
the presence of blood in stool
• iFOBT
• The presence of blood in the stool can be an early sign
• FIT
of benign or malignant gastrointestinal diseases –
colorectal cancer
• Aged 50 years and up must have annual regular FOBT
screening

FECAL OCCULT BLOOD TEST (FOBT)

• A simple, inexpensive, and most frequently performed
chemical screening test on stool specimens
• Occult blood means that you can't see it with the naked
eye. Blood in the stool means there is likely some kind
of bleeding in the digestive tract.
• Blood in the stool may also be a sign of colorectal
cancer, a type of cancer that starts in the colon or
rectum.
• Check stool samples for hidden (occult) blood
• May indicate colon cancer or polyps in the colon or
rectum — though not all cancers or polyps bleed.
Typically, occult blood is passed in such small amounts
that it can be detected only through the chemicals used
in a fecal occult blood test.
• If blood is detected through a fecal occult blood test,
additional tests may be needed to determine the source SUBSTANCES THAT CAUSE FALSE POSITIVE REACTIONS
of the bleeding. ARE:
• The fecal occult blood test can only detect the presence • Drugs - Boric acid, Bromides, Colchicine, Iodine
or absence of blood — it doesn't indicate potential • Animals Foods – foods like meat including processed
sources of bleeding meats and liver in diet that contain hemoglobin,
myoglobin, and certain enzymes
PURPOSE • Vegetables and fruits with peroxidase activity -
• Help determine if you might have an underlying cauliflower, turnips, broccoli, horseradish,
condition such as colon polyps, diverticulosis, radishes/cantaloupe, mushrooms, apples, bananas
hemorrhoids, an ulcer, an inflammatory bowel disease
such as ulcerative colitis, or colorectal cancer. Each of SUBSTANCES THAT CAUSE FALSE NEGATIVE REACTIONS
these (and others) can cause bleeding in your digestive ARE:
tract that ends up in your stool. • Ascorbic acid (Vitamins C)
• This test is not diagnostic but instead shows that • Iron supplements that contain Vitamin C
additional testing is needed.
OTHER FACTORS AFFECTING TEST RESULTS ARE:
PRINCIPLE • Bleeding hemorrhoids
• Detect occult blood is based on the pseudo-peroxidase • Collection of specimens during menstruation
activity of hemoglobin and its derivatives • Hematuria (blood in urine)
• Indicator – benzidine • Some long-distance runners
o toluidine
o guaiac or aminophenazone

PATIENT PREPARATION
The patient should not consume in at least 3 days:
o Rare and lightly cooked meats, especially
o Cauliflower, turnips, broccoli, horseradish,
radishes/cantaloupe
o Iron-rich supplements
o Vit C over 250 mg per day
o 7 days for aspirin and other inflammatory drugs
o Excessive amounts of alcoholic drinks

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