To achieve this unit a learner must:

OUTCOMES
1. Demonstrate the use of reagent strip and interpret results based on the time required.
UNIT 2. Demonstrate the use of clinitest and interpret results.

ANALYTICAL PHASE

Materials

For reagent strips:

Urine (freshly voided or random specimen, preferably first morning specimen)

N-Multistix SG reagent strip

For reagent tablets:

Urine (freshly voided or random specimen
Clinitest tablet

Procedure

For Reagent Strips:

 Completely immerse all reagent areas of the strip in fresh, well-mixed uncentrifuged urine
and remove immediately.
 Tap edge of strip against the side of urine container to remove excess urine. Hold the strip
in a horizontal position to prevent possible mixing of chemicals from adjacent reagent area
and/or soiling of hands with urine.
 Compare test areas closely with corresponding color charts on the bottle label at the times
specified. Hold the strip closely to color blocks and match carefully.

Certain colors continue to become more intense for a short time and then fade. pH and protein
may be read at any time up to one minute after dipping. Glucose should be read at 10 seconds for
qualitative results. The ketone test is read at 15 seconds, bilirubin at 20 seconds, blood at 30 seconds, nitrite
at 30 seconds of immediately following blood portion and urobilinogen at one minute. The Color Chart is
considered to be the “standard” for the colorimeter tests for urine incorporated in N-Multistix SG.

For reagent tablets:

CLINITEST
Clinitest Reagent Tablets

These tablets are quick, simple semi-quantitative tests for estimating sugar urine. They are
an adaptation of the alkaline copper reduction test in self-heating tablet form. The tablet contains
copper sulfate, sodium hydroxide, sodium carbonate and citric acid.
 Always use accompanying test tube and dropper. Other equipment may give incorrect
results.

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 To achieve this unit a learner must:

OUTCOMES
UNIT 1. Identify urinary sediments microscopically under low power and high power objective.
2. Report urinary sediments in correlation with the other parameters of urine.

ANALYTICAL PHASE

Materials

Urine (freshly voided or random specimen, preferably first morning specimen)

Slides and cover slips

Microscope

Procedure

 Thoroughly mix your urine specimen.

 Pour 10 mL of the urine into a centrifuge tube or any test tube used for that matter.
 Centrifuge for 5 mintues at 1500 rpm.
 Pour off the supernatant liquid. Sufficient urine remains in the tube to suspend the
sediment.
 Shake the tube and transfer a drop on the slide. The drop should not overrun the cover slip
and there should be no air bubbles since both tend to alter the results.
 Examine under the microscope using LPO first the HPO.<
 Report the structures seen as follows:
o Casts: total count in 20 low power fields (LPO)
o Erythrocytes and Leukocytes: report in form of range per high power field
(HPO)
o Epithelial cells: report as occasional (+), few (+), moderate (++), many (+++) or
abundant (++++), under LPO and HPO of the specific type encountered.
o Crystals: report as in epithelial cells
o Microorganisms: report as occasional (+), few (+), moderate (++), many (+++)
or abundant (++++) under LPO
o Foreign substances: identify and report the presence under LPO and HPO

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 To achieve this unit a learner must:

OUTCOMES
UNIT 1. Identify the physical characteristics of feces based on its color, odor, and consistency.
2. Demonstrate the procedure of chemical tests for determination of fecal constituents.
3. Report fecal constituents.

Sudan III in 95% alcohol

Applicator sticks
Slides and cover slips
Pea sized stool

4. Split fat procedure

36% acetic acid
Sudan III
Applicator sticks
Slides and cover slips
Pea sized stool

Procedure

A. PHYSICAL EXAMINATION
 Pass out stool in a bedpan lined with newspaper. Avoid admixture of urine, fibers, dirt,
gauze threads and tissue papers.
 By means of an applicator stick, get a pea-size or marble-size of stool from the mid-
portion. If a portion is mucoid or bloody, collect from that portion.
 In case of liquid stool, collect 10ml or 1/3 full of an eight ounce container.
 Examine the specimen immediately.
 Note the color, odor and consistency.

B. CHEMICAL EXAMINATION

1. Fecal occult blood test

 Mix 5 ml of hydrogen peroxide with 5 ml of guiac solution.

 Make the stool specimen acidic using glacial acetic acid (test using litmus paper).
 Apply a small portion of stool in the filter paper.
 Add 2-3 drops of the prepared gum guiac and hydrogen peroxide solution.
 A blue color will appear in the filter paper in the presence of blood.

2. Acid Steatocrit Procedure

 0.5g of feces from a spot collection is diluted 1 to 4 with deionized water.
 Vortex for 2 minutes to homogenize the specimen.
 A volume of 5 N perchloric acid equal to 20% of the homogenate volume is added
and the mixture is then vortexed for 30 seconds. Confirm the pH to be <1.
 Place the acid-homogenate mixture in 75 microliter plain hematoctir capillary tube.
Seal the end with wax.

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 To achieve this unit a learner must:

OUTCOMES
UNIT 1. Identify the physical characteristics of feces based on its color, odor, and consistency.
2. Demonstrate the procedure of chemical tests for determination of fecal constituents.
3. Report fecal constituents.

 The capillary tube is centrifuged horizontally at 13000 rpm for 15 minutes in a

microhematocrit centrifuge. This separates fat as an upper layer overlying a solid
fecal fat.
 The length of the fat and solid layers are measured using a magnifying lens.
 Calculate the acid steatocrit in percent.
 Calculate the fecal fat in grams per 24 hours.

C. MICROSCOPIC EXAMINATION

1. Methylene blue procedure for fecal leukocytes

 Place mucus or a drop of liquid stool on a stool.
 Add two drops Loffler methylene blue.
 Mix with a wooden applicator stick.
 Allow to stand 2-3 minutes.
 Examine for neutrophils under high power.

2. Muscle fiber procedure

 Emulsify a small amount of stool in two drops of 10% eosin in alcohol.
 Coverslip and let stand for 3 minutes.
 Examine under high power field for 5 minutes.
 Count the number of undigested fibers.

3. Neutral fat stain procedure

 Homogenize one part stool with two parts water.
 Mix emulsified stool with one drop 95% ethyl alcohol on slide.
 Add two drops saturated Sudan III in 95% ethanol.
 Mix and coverslip.
 Examine under high power field.
 Count orange droplets per high power field.

4. Split fat stain procedure

 Mix emulsified stool with one drop of 36% acetic acid.
 Add two drops saturated Sudan III.
 Mix and coverslip.
 Heat gently almost to boiling.
 Examine under high power.
 Count and measure the orange droplets per high power field.

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