ACCUPROF LABORATORY AND DIAGNOSTIC CENTER

Clinical Microscopy and Parasitology Technical Procedure Manual

ROUTINE URINALYSIS

Definition
A urinalysis is a group of manual and/or automated qualitative and semi-quantitative tests
performed on a urine sample. A routine urinalysis usually includes the following tests: color,
transparency, specific gravity, pH, protein, glucose, ketones, blood, bilirubin, nitrite,
urobilinogen, and leukocyte esterase. Some laboratories include a microscopic examination of
urinary sediment with all routine urinalysis tests. If not, it is customary to perform the
microscopic exam, if transparency, glucose, protein, blood, nitrite, or leukocyte esterase is
abnormal.

Purpose

Routine urinalyses are performed for several reasons:

 general health screening to detect renal and metabolic diseases

 diagnosis of diseases or disorders of the kidneys or urinary tract
 monitoring of patients with diabetes

In addition, quantitative urinalysis tests may be performed to help diagnose many specific
disorders, such as endocrine diseases, bladder cancer, osteoporosis, and porphyrias.
Quantitative analysis often requires the use of a timed urine sample. The urinary microalbumin
test measures the rate of albumin excretion in the urine using laboratory tests. This test is
used to monitor the kidney function of persons with diabetes mellitus. In diabetics, the
excretion of greater than 200 μg/mL albumin is predictive of impending kidney disease.

Specimen Collection
A urine sample is collected in an unused disposable plastic cup with a tight-fitting lid. A
randomly voided sample is suitable for routine urinalysis, although the urine that is first
voided in the morning is preferable because it is the most concentrated. The best sample for
analysis is collected in a sterile container after the external genitalia has been cleansed
using the midstream void (clean-catch) method. This sample may be cultured if the laboratory
findings indicate bacteruria.
To collect a sample using the clean-catch method:

 Females should use a clean cotton ball moistened with lukewarm water (or antiseptic wipes
provided with collection kits) to cleanse the external genital area before collecting a
urine sample. To prevent contamination with menstrual blood, vaginal discharge, or germs
from the external genitalia, they should release some urine before beginning to collect
the sample.

 Males should use a piece of clean cotton moistened with lukewarm water or antiseptic
wipes to cleanse the head of the penis and the urethral meatus (opening). Uncircumcised
males should draw back the foreskin. After the area has been thoroughly cleansed, they
should use the midstream void method to collect the sample.

 For infants, a parent or health care worker should cleanse the baby's outer genitalia and
surrounding skin. A sterile collection bag should be attached to the child's genital area
and left in place until he or she has urinated. It is important to not touch the inside
of the bag, and to remove it as soon as a specimen has been obtained.
Urine samples can also be obtained via bladder catheterization, a procedure used to collect
uncontaminated urine when the patient cannot void. A catheter is a thin flexible tube that a
health care professional inserts through the urethra into the bladder to allow urine to flow
out. To minimize the risk of infecting the patient's bladder with bacteria, many clinicians
use a Robinson catheter, which is a plain rubber or latex tube that is removed as soon as the
specimen is collected. If urine for culture is to be collected from an indwelling catheter, it
should be aspirated (removed by suction) from the line using a syringe and not removed from
the bag in order to avoid contamination.
Suprapubic bladder aspiration is a collection technique sometimes used to obtain urine from
infants younger than six months or urine directly from the bladder for culture. The doctor
withdraws urine from the bladder into a syringe through a needle inserted through the skin.

Specimen Collection and Transportation Guidelines

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As with any type of laboratory specimen, there are certain criteria that need to be met for
proper collection and transportation of urine specimens. This will ensure proper stability of
the specimen and more accurate test results.

 All patients should avoid intense athletic training or heavy physical work before the
test, as these activities may cause small amounts of blood to appear in the urine.

 Over two dozen drugs are known to interfere with various chemical urinalysis tests. These
include: ascorbic acid, chlorpromazine, L-dopa, nitrofurantoin (Macrodantin, Furadantin),
penicillin, phenazopyridine (Pyridium), rifampin (Rifadin), tolbutamide.

 To minimize sample contamination, women who require a urinalysis during menstruation

should insert a fresh tampon before providing a urine sample.

 All urine collection and/or transport containers should be clean and free of particles or
interfering substances.

 The collection and/or transport container should have a secure lid and be leak-resistant.
Leak-resistant containers reduce specimen loss and healthcare worker exposure to the
specimen while also protecting the specimen from contaminants.

 It is good practice to use containers that are made of break-resistant plastic, which is
safer than glass.

 The container material should not leach interfering substances into the specimen.

 Specimen containers should not be reused.

 There is a need to use an amber colored container for specimens being assayed for light
sensitive analytes, such as urobilinogen and porphyrins. The colorant prevents the
degradation of certain analytes.

 Proper labeling should be applied to the collection container or tubes.

 Many urinary constituents are labile, and samples should be tested within one hour of
collection or refrigerated. Samples may be stored at 2–8°C for up to 24 hours for
chemical urinalysis tests; however, the microscopic examination should be performed
within four hours of collection, if possible.

 The preservatives that are used to prevent loss of glucose and cells may affect
biochemical test results. The use of preservatives should be avoided whenever possible in
urine tests.

Urine Specimen and Handling Guidelines

 Labels. Include the patient name and identification on labels. Make sure that the
information on the container label and the requisition match. If the collection container
is used for transport, the label should be placed on the container and not on the lid,
since the lid can be mistakenly placed on a different container. Ensure that the labels
used on the containers are adherent under refrigerated conditions.

 Volume. Ensure that there is sufficient volume to fill the tubes and/or perform the
tests. At least 60 mL of urine must be submitted. Under filling or overfilling containers
with preservatives may affect specimen-to-additive ratios.

 Collection Date and Time. Include collection time and date on the specimen label. This
will confirm that the collection was done correctly. For timed specimens, verify start
and stop times of collection. Document the time at which the specimen was received in the
laboratory for verification of proper handling and transport after collection. Urine
specimens should be examined within 1 hour after collection, or else refrigerated.

 Collection Method. The method of collection should be checked when the specimen is
received in the laboratory to ensure the type of specimen submitted meets the needs of
the test ordered. An example of an optimum specimen/test match would be a first morning
specimen for urinalysis and microscopic examination.

Standardization

The laboratory must follow a standard protocol of specimen collection and processing which is
essential for accuracy and precision in routine urinalysis. The goal of standardization is to
reduce the ambiguity and subjectivity inherent in the procedure itself. Standardization begins

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when the specimen arrives in the laboratory, with all the specimens being processed according
to strict protocol.

 Volume of Urine Examined. The volume of urine required depends on the number of tests
required. However, a consistent volume of 12 mL should be processed through the routine
urinalysis procedure.

 Length and Speed of Centrifugation. The well-mixed urine is centrifuged in a tube at a

set speed of 3000 rpm for 5 minutes. Longer centrifugation times and higher relative
centrifugal force, although useful in recovering cells, are apt to break up cellular
casts.

 Volume of Sediment Examined. A specific volume of urine sediment must be examined. In

this case, 20 µL is required.

Specimen Processing
Urine specimens must be processed within 1 hour of collection. In the event that testing is
not possible within the allowed time frame, appropriately labelled samples can be stored at a
controlled temperature (2 –8 Celsius) until testing is possible.
Routine urinalysis consists of three testing groups: physical characteristics, biochemical
tests, and microscopic evaluation.
A. General steps
1. Label a urine test tube with patient’s ID no. and name.
2. Invert urine to mix and pour 12 mL of urine into the test tube.
3. Determine color, transparency (clarity).
4. Perform reagent strip testing. Determine pH, Specific gravity, glucose and albumin. Use
unspun, well-mixed urine equilibrated to room temperature. Follow exact timing. Report
results using standardized terminology.
5. Centrifuge specimen for 5 minutes at 3,000 rpm.
6. Decant the supernatant (liquid layer).
7. Microscopic analysis of sediment: Thoroughly mix sediment.
8. Using a pipette, transfer 20 µL of suspended sediment to the center of a clean glass
slide.
9. Place slide on microscope stage for observation.

B. Physical tests
The physical tests measure the color, transparency (clarity), and specific gravity of a
urine sample. In some cases, the volume (daily output) may be measured. Color and
transparency are determined from visual observation of the sample.
1. Color. Normal urine is straw yellow to amber in color. Abnormal colors include bright
yellow, brown, black (gray), red, and green. These pigments may result from
medications, dietary sources, or diseases. For example, red urine may be caused by
blood or hemoglobin, beets, medications, and some porphyrias. Black-gray urine may
result from melanin (melanoma) or homogentisic acid (alkaptonuria, a result of a
metabolic disorder). Bright yellow urine may be caused by bilirubin (a bile pigment).
Green urine may be caused by biliverdin or certain medications. Orange urine may be
caused by some medications or excessive urobilinogen. Brown urine may be caused by
excessive amounts of prophobilin or urobilin (a chemical produced in the intestines).
2. Transparency. Normal urine is transparent. Turbid (cloudy) urine may be caused by
either normal or abnormal processes. Normal conditions giving rise to turbid urine
include precipitation of crystals, mucus, or vaginal discharge. Abnormal causes of
turbidity include the presence of blood cells, yeast, and bacteria.
3. Specific Gravity. The specific gravity of urine is a measure of the concentration of
dissolved solutes, and it reflects the ability of the kidneys to concentrate the urine
(conserve water). Specific gravity is usually measured by determining the refractive
index of a urine sample (refractometry) or by chemical analysis. Specific gravity
varies with fluid and solute intake. It will be increased (above 1.035) in persons with
diabetes mellitus and persons taking large amounts of medication. It will also be
increased after radiologic studies of the kidney owing to the excretion of x-ray
contrast dye. Consistently low specific gravity (1.003 or less) is seen in persons with
diabetes insipidus. In renal failure, the specific gravity remains equal to that of
blood plasma (1.008–1.010) regardless of changes in the patient's salt and water
intake. Urine volume below 400 mL per day is considered oliguria, and may occur in

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persons who are dehydrated and those with some kidney diseases. A volume in excess of 2
liters (slightly more than 2 quarts) per day is considered polyuria; it is common in
persons with diabetes mellitus and diabetes insipidus.
C. Biochemical tests
Biochemical testing of urine is performed using dry reagent strips, often called dipsticks.
A urine dipstick consists of a white plastic strip with absorbent microfiber cellulose pads
attached to it. Each pad contains the dried reagents needed for a specific test. The person
performing the test dips the strip into the urine, lets it sit for a specified amount of
time, and compares the color change to a standard chart.
Additional tests are available for measuring the levels of bilirubin, protein, glucose,
ketones, and urobilinogen in urine. In general, these individual tests provide greater
sensitivity; they therefore permit detection of a lower concentration of the respective
substance. A brief description of the most commonly used dry reagent strip tests follows.
1. pH. A combination of pH indicators (methyl red and bromthymol blue) react with hydrogen
ions (H+) to produce a color change over a pH range of 5.0 to 8.5. pH measurements are
useful in determining metabolic or respiratory disturbances in acid-base balance. For
example, kidney disease often results in retention of H+ (reduced acid excretion). pH
varies with a person’s diet, tending to be acidic in people who eat meat but more
alkaline in vegetarians. pH testing is also useful for the classification of urine
crystals.
2. Protein. Based upon a phenomenon called the “protein error of indicators”, this test
uses a pH indicator, such as tetrabromphenol blue, that changes color (at constant pH)
when albumin is present in the urine. Albumin is important in determining the presence
of glomerular damage. The glomerulus is the network of capillaries in the kidneys that
filters low molecular weight solutes such as urea, glucose, and salts, but normally
prevents passage of protein or cells from blood into filtrate. Albuminuria occurs when
the glomerular membrane is damaged, a condition called glomerulonephritis.
3. Glucose. The glucose test is used to monitor persons with diabetes. When blood glucose
levels rise above 160 mg/dL, the glucose will be detected in urine. Consequently,
glycosuria may be the first indicator that diabetes or another hyperglycemic condition
is present. The glucose test may be used to screen newborns for galactosuria and other
disorders of carbohydrate metabolism that cause urinary excretion of a sugar other than
glucose.
4. Ketones. Ketones are compounds resulting from the breakdown of fatty acids in the body.
These ketones are produced in excess in disorders of carbohydrate metabolism,
especially Type 1 diabetes mellitus. In diabetes, excess ketoacids in the blood may
cause life-threatening acidosis and coma. These ketoacids and their salts spill into
the urine, causing ketonuria. Ketones are also found in the urine in several other
conditions, including fever; pregnancy; glycogen storage diseases; and weight loss
produced by a carbohydrate-restricted diet.
5. Blood. Red cells and hemoglobin may enter the urine from the kidney or lower urinary
tract. Testing for blood in the urine detects abnormal levels of either red cells or
hemoglobin, which may be caused by excessive red cell destruction, glomerular disease,
kidney or urinary tract infection, malignancy, or urinary tract injury.
6. Bilirubin. Bilirubin is a breakdown product of hemoglobin. Most of the bilirubin
produced in humans is conjugated by the liver and excreted into the bile, but a very
small amount of conjugated bilirubin is reabsorbed and reaches the general circulation
to be excreted in the urine. The normal level of urinary bilirubin is below the
detection limit of the test. Bilirubin in the urine is derived from the liver, and a
positive test indicates hepatic disease or hepatobiliary obstruction.
7. Specific gravity. Specific gravity is a measure of the ability of the kidneys to
concentrate urine by conserving water.
8. Nitrite. Some disease bacteria, including the lactose-positive Enterobactericeae,
Staphylococcus, Proteus, Salmonella, and Pseudomonas are able to reduce nitrate in
urine to nitrite. A positive test for nitrite indicates bacteruria, or the presence of
bacteria in the urine.
9. Urobilinogen. Urobilinogen is a substance formed in the gastrointestinal tract by the
bacterial reduction of conjugated bilirubin. Increased urinary urobilinogen occurs in
prehepatic jaundice (hemolytic anemia), hepatitis, and other forms of hepatic necrosis
that impair the circulation of blood in the liver and surrounding organs. The

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urobilinogen test is helpful in differentiating these conditions from obstructive

jaundice, which results in decreased production of urobilinogen.
10. Leukocytes. The presence of white blood cells in the urine usually signifies a urinary
tract infection, such as cystitis, or renal disease, such as pyelonephritis or
glomerulonephritis.
D. Microscopic examination
A urine sample may contain cells that originated in the blood, the kidney, or the lower
urinary tract. Microscopic examination of urinary sediment can provide valuable clues
regarding many diseases and disorders involving these systems.
The presence of bacteria or yeast and white blood cells helps to distinguish between a
urinary tract infection and a contaminated urine sample. White blood cells are not seen if
the sample has been contaminated. The presence of cellular casts (casts containing RBCs,
WBCs, or epithelial cells) identifies the kidneys, rather than the lower urinary tract, as
the source of such cells. Cellular casts and renal epithelial (kidney lining) cells are
signs of kidney disease.
The microscopic examination also identifies both normal and abnormal crystals in the
sediment. Abnormal crystals are those formed as a result of an abnormal metabolic process
and are always clinically significant. Normal crystals are formed from normal metabolic
processes; however, they may lead to the formation of renal calculi, or kidney stones.
Steps:
1. Scan sample using the low power (10X) objective.

2. Determine frequency and distribution of formed elements. They should be uniform. Be

certain to scan areas near edge of the cover slip. Due to the mechanical motion
involved in “cover-slipping” the sample, heavier elements (i.e. casts) tend to disperse
towards the edge of cover slip. Observe for “motile” cells. Trichomonas species can be
easily spotted using this objective.

3. Epithelial cells with the exception of renal tubular and transitional cells are
counted. Casts are also counted and reported as the average of the fields per low power
field (LPF).

4. Identify and classify epithelial cells according to the following:

o squamous
o renal tubular
o transitional
Casts and crystals are identified (if present) according to structural characteristics
or cellular elements contained:
o Cellular casts (i.e. Epithelial, RBC, WBC, bacterial, or mixed)
o Coarsely granular
o Finely granular
o Broad cast
o Waxy cast
o Hyaline
NOTE: For identification purposes, it is advisable to switch to higher magnification
objective (40X); however, average counts must be performed using the low objective
(10X).

5. Finally, grade presence of mucus as: slight, moderate, or marked.

6. While the larger structures are identified using the 10X objective, smaller cellular
elements such as WBC’s, RBC’s, and yeast are counted and reported as the average of the
fields counted per High power field (HPF) objective or 40X. Bacteria are reported as
few, moderate, or many (see below).

7. WBC clumps may be reported as such including the number of clumps observed.

8. Extremely bloody sediments may obscure the presence of other elements. In such case,
mix equal amounts of sediment with 5% Acetic acid (i.e. 2 drops sediment + 2 drops
acid). This action will result in a dilution factor of 2; therefore, counts must be
multiplied by 2 before reporting. The addition of acetic acid causes red blood cells to
hemolyse and accentuates the leukocytes nuclei.

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9. Different crystals are encountered at varying pH levels (acid, neutral, or alkaline

urine). Most crystals are not clinically significant. Findings of abnormal crystals
(cysteine, leucine, tyrosine), are rare and should be confirmed in the laboratory
before reporting.

Manner of Reporting

 Mucus threads: Rare, Few, Moderate, Plenty

 Amorphous crystals: Rare, Few, Moderate, Plenty
 Epithelial cells: Rare, Few, Moderate, Plenty
 Casts: Average count / LPF; specify type
 Crystals: Rare, Few, Moderate, Plenty; specify type
 WBC: Average count / HPF; Maximum count > 100 HPF
 RBC: Average count / HPF; Maximum count > 100 HPF
 Bacteria/Yeast: Rare, Few, Moderate, Plenty

Normal Results

Normal urine is a clear straw-colored liquid, but may also be slightly hazy. It may contain
some normal crystals as well as squamous or transitional epithelial cells from the bladder,
lower urinary tract, or vagina. Urine may contain transparent (hyaline) casts, especially if
it was collected after vigorous exercise. The presence of hyaline casts may be a sign of
kidney disease, however, when the cause cannot be attributed to exercise, running, or
medications. Normal urine contains a small amount of urobilinogen, and may contain a few RBCs
and WBCs. Normal urine does not contain detectable amounts of glucose or other sugars,
protein, ketones, bilirubin, bacteria, yeast cells, or trichomonads. Normal values used in
many laboratories are given below:

 Glucose: Negative (Quantitative less than 130 mg/day or 30 mg/dL)

 Bilirubin: Negative (Quantitative less than 0.02 mg/dL)
 Ketones: Negative (Quantitative 0.5–3.0 mg/dL)
 pH: 5.0–8.0
 Protein: Negative (Quantitative 15–150 mg/day, less than 10 mg/dL)
 Blood: Negative
 Nitrite: Negative
 Specific gravity: 1.015–1.025
 Urobilinogen: 0–2 Ehrlich units (quantitative 0.3–1.0 Ehrlich units)
 Leukocyte esterase: Negative
 Red blood cells: 0–2 / HPF
 White blood cells: 0–5 / HPF

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STOOL ANALYSIS (FECALYSIS)

Overview
A stool analysis is a series of tests done on a stool (feces) sample to help diagnose certain
conditions affecting the digestive tract. These conditions can include infection (such as from
parasites, viruses, or bacteria), poor nutrient absorption, or cancer.
For a stool analysis, a stool sample is collected in a clean container and then sent to the
laboratory. Laboratory analysis includes microscopic examination, chemical tests, and
microbiologic tests. The stool will be checked for color, consistency, weight (volume), shape,
odor, and the presence of mucus. The stool may be examined for hidden (occult) blood, fat,
meat fibers, bile, white blood cells, and sugars called reducing substances. The pH of the
stool also may be measured. A stool culture is done to find out if bacteria may be causing an
infection.

Visual Observation of the Fecal Sample

It is important to observe the macroscopic appearance of the stool as this can give a clue to
the type of organisms present. Therefore the consistency; formed, unformed or liquid; the
colour and the presence or absence of the exudate are reported. The presence of adult worms
can also be seen in a freshly passed stool e.g. adult stage of Ascaris lumbricoides and
Enterobius vermicularis. Proglottids of Taenia species can also be seen.

Microscopic Examination of Fecal Samples for Parasites

Direct microscopy should be done on all unformed and liquid samples by mixing a small amount
of the specimen in 0.9% sodium chloride solution. This permits detection of trophozoites of
Entamoeba histolytica and Giardia lamblia. It can also provide information on the content of
the stool i.e. the presence of leukocytes and red blood cells.

Principles of Diagnostic Methods for the Identification of Parasites

The principle of the successful identification of faecal parasites is based upon,

1. Measurement. An eyepiece graticule may be used, especially for cyst identification.
2. Morphology. In protozoan cysts, the number of nuclei and the presence of inclusions e.g.
glycogen mass and chromidial bar, aid the identification of protozoa. In trophozoites, the
presence of red cells in amoebae is diagnostic of Entamoeba histolytica and flagella also
aid identification of some protozoan trophozoites.
3. Appearance. In helminth eggs, the shape of the egg, the thickness of the shell, the colour
of the ovum and the presence or absence of features such as an operculum, spine or hooklets
are diagnostic pointers to the identity of the parasite.
4. Stains also aid in identification of the parasite. The addition of iodine highlights the
internal structure of cysts and helps distinguish between vegetative matter and cysts.
Permanently stained fecal smears are useful for demonstrating the nuclear patterns of
cysts.

Procedure

1. Place a small amount of feces on a microscope slide.

2. Add a drop of liquid to the feces and mix thoroughly. The type of liquid added depends on
what you hope to accomplish with the technique. If examining a liquid fecal sample for the
presence of protozoan trophozoites (live active protozoa) then use saline (if any extra
liquid is needed). If looking for helminth eggs and protozoan cysts in a small sample then
either water or iodine may be used.
3. Cover with a cover slip. Move the cover slip around until it lays flat.
4. Examine the slide using the 10X objective, and then go over it with the 40X objective.

Reporting of Parasites

Ideally, the presence of all parasites should be reported, whether they are pathogens or non-
pathogens. This particularly applies to the presence of cysts. The stage of the parasite
should always be reported. For the protozoa, whether cysts or trophozoites are present; the
stage of larvae as in Strongyloides; and whether adult stages or eggs of helminths are
present.

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If no parasite is found, report as “Negative for intestinal parasite.”

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FECAL OCCULT BLOOD TEST

Overview
The occult blood test is a rapid and convenient qualitative method for detecting fecal occult
blood. Occult is word meaning “hidden”. Blood can be present in a stool sample, but due to the
digestive process, will not retain its bright red color. The occult blood tests detect excess
blood loss which may have significance when related to certain diseases such as colorectal
cancer. A positive test usually indicates blood in excess of normal and should be followed up
medically. A negative test usually indicates that no blood, in excess of normal, is apparent
in the fecal specimen tested. The accuracy of the test depends upon the status of the patient
at the time the specimen is taken and may be affected by interfering substances.
The occult blood is recommended for use as a diagnostic aid during routine physical
examination, when hospital patients are first admitted, to monitor for bleeding in patients
recuperating from surgery and other conditions, and in screening programs for colorectal
cancer. It is not a test for colorectal cancer or any other specific disease. It is used as a
qualitative aid to the diagnosis of various gastrointestinal conditions which manifest
themselves by the presence of fecal occult blood.

Principle of the Test

The test consists of a special guaiac impregnated paper. A smear from a stool sample is
applied to one side of the paper, the paper is turned over and a special developer is added.
The developer will react with hemoglobin released from lysed red blood cells resulting in the
formation of blue color if blood is present. The test reaction is based on the oxidation of
guaiac by hydrogen peroxide to a blue-colored compound.
Hemoglobin, if present in the fecal specimen, acts as a pseudo-peroxidase material. It
catalyzes the oxidation of alpha guaiaconic acid (active component of guaiac paper) by
hydrogen peroxide (active component of the developer) to form a highly conjugated blue quinone
compound. Appearance of any blue color on the specimen area of the slide is an indication of
the presence of occult blood.

Patient Preparation

If possible the patient should be placed on a meat-free low-peroxidase diet to reduce the
possibility of false positive indications. This special diet should be started two days before
testing and continued through the testing period. Such a diet may help reduce the number of
false positive results. It also provides roughage that may help uncover silent lesions which
may bleed intermittently and may increase the rate of true positive reactions. The recommended
diet will also increase the likelihood of a soft stool for greater ease in obtaining fecal
samples by wiping.

Suggested Diet
Avoid:
1. Red or rare meat, and the following raw vegetables and fruits: broccoli, turnips,
horseradish, cauliflower, red radishes, parsnips and cantaloupe.
2. Vitamin C in excess of 250 mg per day.
3. Aspirin and anti-inflammatory drugs which may cause gastrointestinal irritation for 7 days
prior to and during the test period.
4. Iron supplements.
Try to eat:
1. Cooked vegetables and fruits, especially lettuce, spinach, and corn.
2. Prunes, grapes, plums and apples.
3. Peanuts, popcorn and bran cereals.
4. Well-cooked fowl and canned tuna fish.
If any of the above dietary restrictions and recommendations are known to cause discomfort,
patients should be instructed to inform their physician. The patient should always consult the
physician before discontinuing or interrupting any prescription medications.

Specimen Collection
The specimen required is a small stool sample which should be applied as a very thin smear
onto both windows of the slide. Slides may be developed immediately after specimen application
or may be stored and developed up to 8 days after specimen application. Once they have been

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prepared with a specimen, keep the slides away from heat and light. The work area should be
kept clean and free of blood to avoid accidental contact of blood with the slides.
Patients experiencing hemorrhoidal bleeding, having a menstrual period, or bleeding from the
nose, gums, etc. should delay testing for at least 48 hours from the time that all such
bleeding has stopped. To increase the chances of detecting intermittent gastrointestinal
bleeding, it is recommended that stool samples be collected from three consecutive bowel
movements and that two smears be made from two different areas of each bowel movement,
especially from darkened or discolored areas of the feces. Excessive GI bleeding may result in
black, tarry stools.

Procedure

1. Write the patient information from the front flap of the slide onto your report form.
2. Turn the slide over and open the flap to expose the test area. NOTE: The slides have fecal
material on them, handle with care.
3. Apply two drops of developer solution to each smear in the Specimen Test Area.
4. Read results within 30 to 60 seconds. Any trace of blue color is positive for occult blood.
Color begins to fade after 2 to 4 minutes.
5. Develop the performance control only after specimen tests have been completed and
interpreted. Apply one drop of developer solution to the Performance control Line. A blue
color should appear within 30 seconds when the reagent test paper and developer are
performing according to specifications.
6. Record the results of the patient and the performance control on the report form.

Interpretation
Record results as “Positive” or “Negative. Any trace of blue color within the specimen
application area is positive for occult blood. The “control” area of the slide is at the
bottom of the slide. The positive control has a “+” and must be blue, the negative control is
a “-“ and must remain colorless.

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