Clin Path Lab 6 Urinalysis Part 2

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ROUTINE URINALYSIS USTMED ’07 Sec C - AsM Components of a Urinalysis Physical Examination of Urine

1. color 2. transparency 3. specific gravity 4. pH

Chemical Examination of Urine (using a multi-parameter reagent strip) 1. Protein 2. Glucose 3. Ketone 4. Bilirubin 5. Hemoglobin 6. Urobilinogen 7. Nitrite 8. Leukocyte esterase

Microscopic Examination of Sediments Collection and Handling of Urine Specimen

Urine must be collected in a clean, dry container. Disposable containers are recommended in order to reduce bacterial contamination.

Label the specimen container w/ the patient’s name, date and time of collection. Place the label on the container and not on the lid.

Urine collected should be sent to the lab as soon as possible and should be tested w/in 1 hr. Refrigeration is necessary if the urine cannot be delivered immediately to the laboratory or cannot be tested w/in 1 hr. The addition of a chemical preservative can also be done.

Types of urine specimens 1. random specimen

– most commonly received specimen 2. First morning urine

- the ideal screening specimen

- used for routine screening, pregnancy tests and for determining orthostatic proteinuria

3. Fasting Specimen (Second Morning Specimen)

- second voided urine specimen used for glucose monitoring

4. 24-hour specimen

- timed specimen is used to determine concentration of a particular substance

- Instruction:

o Day 1 – 7am - patient voids and discards specimen - patient collects all urine for the next 24 hours o Day 2 – 7am - patient voids and adds

this urine to the previously collected urine

5. Catheterized Specimen - used for bacterial culture 6. Midstream Clean-Catch Specimen

- an ideal specimen for routine screening and for bacterial culture

- patient is instructed to cleanse thoroughly the genitalia and is asked to collect the midstream portion of urine. When collecting, patient should be instructed to separate the labia in females or retract the foreskin in uncircumcised males. 7. Suprapubic Aspiration

- used for bacterial culture and cytology

- sterile needle is introduced into the bladder to collect sample that is free of contaminants

8. Pediatric Specimen

- soft, clear plastic bag w/ adhesive is attached to the genital portion

PHYSICAL EXAMIANTION OF URINE Odor

- usually not a part of routine urinalysis

- ammoniacal odor of urine is due to breakdown of urea

Odor Pathologic Nonpathologic

Ammonia UTI Old urine

Sweet Diabetes mellitus Starvation, dieting, strenuous exercise, vomiting, diarrhea

Mousy Phenylketonuria

Maple syrup Maple syrup disease

Distinctive Garlic, onions,

asparagus

Appearance and Color of Urine

Apprnce Cause Remarks

Colorless Very dilute urine Polyuria, diabetes insipidus Cloudy Phosphates, carbonates

Urates, uric acid Leukocytes Red cells (“smoky”) bacteria, yeasts spermatozoa prostatic fluid mucin, mucous threads calculi, “gravel” clumps, pus, tissue fecal contamination radiographic dye

Soluble in dilute acetic acid Dissolve in 60oC and in alkali Insoluble in dilute acetic acid Lyse in dilute acetic acid Insoluble in dilute acetic acid Insoluble in dilute acetic acid May be flocculent

Phosphates, oxalates Rectovesical fistula In acid urine Milky Many neutrophils

(pyuria) Fat Lipiduria,opalascent Chyluria, milky Emulsified paraffin

Insoluble in dilute acetic acid

Nephrosis, crush injury – soluble in ether Lymphatic obstruction –

soluble in ether Vaginal creams Yellow Acriflavine Green Fluorescence

Yellow-orange Concentrated urineUrobilin in excess Bilirubin

Dehydration, fever No yellow foam

Yellow foam, if sufficient bilirubin

Yellow-green Bilirubin-biliverdin Yellow foam

Yellow-brown Bilirubin-biliverdin “beer” brown, yellow foam

Red Hemoglobin

Erythrocytes Myoglobin Porphyrin Fuscin, aniline dye Beets

Menstrual contamination

Positive reagent strip for blood

Positive reagent strip for blood

Positive reagent strip for blood

May be colorless Foods, candy

Yellow alkaline, genetic Clots, mucus

Red-purple Porphyrins May be colorless Red-brown Erythrocytes Hemoglobin on standing Methemoglobin Myoglobin Bilifuscin (dipyrrole) Acid pH Muscle injury Result of unstable hemoglobin

Brown-black MethemoglobinHomogentisic acid Melanin Blood, acid pH On standing, alkaline On standing, rare Blue-green Indicans Pseudomonas infections Chlorophyll

Small intestine infection

Mouth deodorants

Color

Normal urine color – Yellow (due to urochrome)

Color Pathologic NonPatho Drugs

Orange Bilirubin Rhubarb

Carrots Phenazopyridine Dark Yellow Bilirubin

Urobilin CarrotsConcentrated Fluorescein Green Oxidized

bilirubin Biliverdin Pseudomonas

Vit B complx Dithiazanine Nitorfurans Phenol Red or Pink RBCs Hemoglobin Myoglobin Porphyrins Beets Benzene Acetophenetidin Phenindione Brown or

Black Biliary pigments Melanin Hmogentisic acid Rhubarb Phenol derivatives Methyldopa Levodopa Pale Yellow Diabetes

mellitus Diabetes insipidus

Large fluid

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Transparency (clarity) Normal appearance

- freshly voided urine is usually clear

- white cloudiness may be caused by precipitation of amorphous phosphates and carbonates

Causes of turbidity

- presence of phosphates, carbonates, crystals - red blood cells, leukocytes, epithelial cells, etc. Specific Gravity

- defined as the density of a substance compared with the density of a similar volume of distilled water at a similar temperature

- used to assess kidney’s ability for reabsorption - Normal values = 1.015-1.025

ROUTINE URINALYSIS PROCEDURE Urine in specimen container Transfer urine in a clean tube

Step 1 Physical Examination *note color & transparency

*specific gravity & pH will be done using the Multistix (Step)2 Step 2 Chemical Examination

* use Multistix Reagent Strip and test for glucose, bilirubin, ketones, specific gravity, blood, pH, protein, Urobilinogen, nitrite

& leukocytes Centrifuge for 5 mins

Decant supernatant & place a drop of the sediments onto a slide Step 3 Microscopic Examination

*examine under LPO & HPO and write your findings in the result form

QUANTITATIVE DETERMINATION OF TOTAL PROTEIN IN URINE A. Pyrogallol Red-Molybdate Method

Clinical Significance:

Urinary protein determinations are important in the evaluation of kidney function. The detection of protein in urine is considered evidence of renal disease and usually of glomerular disease. A minor evaluation of urinary albumin excretion from about 15-30 mg/day is now recognized to be a useful indicator of renal disease.

Principle:

The Total Protein Test for Urine is based on the procedure developed by Watanabe et al which is a dye-binding colorimetric method utilizing pyrogallol red-molybdate complex.

The pyrogallol red is combined w/ molybdenum acid, forming a red cplx w/ maximum abs at 467nm. When this cplx is combined w/ protein in acidic conditions, a blue-purple color develops w/ an inc in abs to 598nm.

Reagents:

1. PRM color reagent

• pyrogallol red-molybdate soln buffered at pH 2.2 2. Microprotein standard

• (50 mg/dl) a soln of bovine serum albumin with preservative

Specimen Collection & Handling:

Urine – random or 24 hr urine collection. Keep specimen cold during collection.

Analyze fresh. Stable frozen at -20oC for up to one year. Procedure:

Unknown Standard Blank

PRM reagent 3.0 ml 3.0 ml 3.0 ml

Serum 0.1 ml -

-Protein Standard - 0.1 ml

-Distilled water - - 0.1 ml

Allow to stand at room temperature for 20 mins

Set wavelength of the photometer at 600 nm and zero with the BLANK. Read and record readings of remaining vials.

Computation: Example-Au = Abs of Unknown Au = 0.16 As = Abs of Standard As = 0.20 Cu = Concentration of Unk Cu = ? Cs = Conc of Standard Cs = 50 mg/dl Formula: Au x Cs = Cu in mg/dl 0.16 x 50 = 40 mg/dl As 0.20

Urine Total Protein (mg/24 hrs or g/24 hrs)

Urine Total Protein = Au x Cs x Volume in dl (24 hr urine) As

To convert mg/dl to g/L: Divide computed value by 100 or multiply by 0.01 Example-Au = 0.12 As = 0.08 Cs – 50 mg/dl Urine volume = 10 dl or 1 L

Urine Total Protein = 0.12 x 50 mg/dl x 10 dl 0.08 = 1.5 x 50 mg/dl x 10 df = 750 mg/24 hrs to convert to g/24 hrs: Divide by 1000 = 0.75 g/24 hrs Normal Values Random urine = 0.01 – 0.14 g/L Urine Total protein = up to 0.15 g/24 h4s B. Microalbumin

An immunological, semi-quantitative determination of mcroalbuminuria up to 100 mg/L

Principle:

Immunological detection of human albumin. The absorbed urine enters a zone on the strip containing a soluble antibody-gold-conjugate which specifically binds to urine albumin. Excess conjugate is retained in a separation zone containing immobilized human albumin so that only the albumin-loaded gold conjugate reaches the detection zone. The color produced (white to red) is directly related to the albumin content of the urine. Cross-reactions with other human proteins such as hemoglobin, transferring, Bence-Jones protein, α1-antitrypsin, acidic-α1-glycoprotein, α-amylase, Tamm-Horsfall protein and retinol-binding protein, as well as with IgG, IgA, human leukocytes and erythrocytes have been found to be <0.5%.

Application:

For early detection and monitoring the course of incipient nephropathy in e.g. diabetics and hypertensive patients. Microalbuminuria denotes as albumin excretion of 20-200 mg/L. Test Component

One test strip contains: Monoclonal antibodies against human albumin (Immunoglobulin G) labeled with colloidal gold: 2.2ug, fixed albumin: 7.7 ug.

Ideal Specimen: First morning urine Results:

Reaction colors lighter than the color block corresponding to approx 20 mg/L albumin indicate a physiological urine albumin conc.

The result is positive (i.e. indicates persistent microalbuminuria) when at least two of the three morning urines tested produce a reaction color corresponding to 20 mg/L (Threshold value of microalbuminuria) or more.

Determination of albumin concentrations above 100 mg/L

In order to determine albumin conc above 100 mg/L, the urine sample can be diluted e.g. by mixing 1 part of urine with 2 parts water. The original albumin concentration is then calculated by multiplying the result by 3.

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L eu co c y tes Nitri te U ro bi li n o gen P ro tei n pH Blo o d S peci fi c Gra vi ty Keto n e B il ir u bi n Gl u c o se

CHEMICAL EXAMINATION OF URINE

COMBISTIX / MULTISTIX Multiple Reagent Strips for Urinalysis Semi-quantitative determination of the following parameters:

Specific gravity Bilirubin

pH Hemoglobin/Myoglobin

Albumin Urobilinogen

Glucose Nitrite

Ketone Esterase

Specimen Collection and Preparation:

Use a freshly voided, well-mixed uncentrifuged urine specimen, collected in a clean container. If testing is not possible one hour after voiding, refrigerate specimen and return it to room temperature before testing. No preservative will prevent deterioration of ketones, bilirubin or urobilinogen, if certain contaminating organisms are present, they may metabolize glucose. Always handle specimen under sanitary conditions. Contamination of urine sample with chlorhexidine antiseptic will give false-positive reading for protein.

Procedure: MUST BE FOLLOWED EXACTLY TO ACHIEVE RELIABLE TEST RESULTS

1. collect FRESH urine specimen in a clean, dry container. Mix well immediately before testing.

2. remove one strip from bottle and replace cap. Completely immerse reagent areas of the strip in FRESH urine and remove immediately to avoid dissolving out reagents

3. While removing, run the edge of the strip against the rim of the urine container to remove excess urine. Hold the strip in a horizontal position to prevent possible mixing of chemicals from adjacent reagent areas/or contaminating the hands with urine.

4. Compare reagent areas to corresponding Color chart on the bottle label at the time specified. Hold strip close to color blocks and match carefully. Avoid laying the strip directly on the Color chart as this will result in the urine soiling the chart

For optimal results, read the ketone test at 15 seconds after dipping; read the bilirubin test at 20 seconds; glucose at 30 seconds; blood at 40 seconds; urobilinogen at 45 seconds; and specific gravity from 45 to 60 seconds after dipping.

I. Ketone (15 secs) 40 seconds nakalagay sa table…???

Principle: Sodium Nitroprusside Rxn

The test is based on the development of colors ranging from buff pink for negative reading to maroon when acetoacetc acid reacts with nitro-prusside. Normally no ketones are present in urine. As little as 0.5-1 mmol/L acetoacetic acid is detectable. Positive results (trace or less) may occur with highly pigmented urine specimens or those containing large amounts of L-Dopa metabolites.

Reagent Composition: Sodium nitroprusside

II. Bilirubin (20 secs) 30 seconds on the table…

Principle: Diazo reaction

Coupling of bilirubin w/ diazotized dichloroaniline in a strongly acid medium yielding various shades of tan.

Reagent Composition:

2,4 dichloroaniline diazonium salt

This test is based on the coupling of biilrubin w/ diazotized dichloroaniline in a strongly acid medium yielding various shades of tan. Normally, no bilirubin is detectable in urine by even the most sensitive tests, so any positive result calls for further investigation. The test has a sensitivity of 7-14 umol/L bilirubin. Colors that do not match the color blocks may result from the presence of bile pigments other than bilirubin, indicating bile pigment abnormalities and suggesting the need for further testing with ICTOTEST reagent tablet, which are 2 to 4 times more sensitive than the strip test. (If the presence of very small conc of bilirubin is suspected, as in early phases of live disease, ICTOTEST should be used.) Specimens from patients receiving large doses of chlorpromazine may give false positive results. Metabolites of drugs such as Pyridium and conc of 1.4 mmol/L or greater may cause false negatives.

III. Glucose (30 secs for quantitative results)

Principle: Glucose oxidase, double sequential enzyme rxn

Glucose oxidase converts glucose to gluconic acid and hydrogen peroxide. Peroxidase then catalyzes the rxn of hydrogen peroxide w/ a potassium iodide (KI) chromogen to form a green to brown color.

Reagent Composition:

Glucose oxidase, peroxidase, potassium iodide, buffer, non-reactive ingredients.

In a double sequential enzyme rxn, glucose oxidase converts glucose to gluconic acid and hydrogen peroxide. Peroxidase then catalyzes the rxn of H2O2 w/ KI chromogen to form a green to brown color. This test yields negative results w/ normal urines. Significant abnormality may be indicated by results of as little as 5 mmol/L if found consistently. High specific gravity, low temperature or moderately high amounts of ketones (4 mmol/L acetoacetic acid or greater) may cause false negatives in specimens containing small amounts of glucose (5 mmol/L). However, the combination of such high ketone levels and low glucose levels is metabolically improbable in screening. Ascorbic acid concentrations of 2.9 mmol/L or greater may cause false negative for specimens containing small amounts of glucose (5 mmol/L). At glucose levels of 60 mmol/L or greater the color may appear uneven. Use the darkest color appearing on the reagent area to interpret the results.

IV. Blood (40 seconds) 60 seconds on the table…

Principle: Peroxidase-like activity of hemoglobin

Catalyzes the reaction of cumene hydroperoxide and tetramethylybenzidine

Note: the appearance of green spots on the reacted reagent area indicates the presence of intact red cells in the urine.

Reagent Composition:

Cumene hydroperoxide, tetramethylbenzidine

This test is based on the peroxide-like activity of hemoglobin w/c catalyzes the rxn of cumene hydroperoxide and 3,3’, 5,5’-tetramethylbenzidine. The resulting color ranges from orange through green to dark blue. Blood is often but not always found in the urine of menstruating females. The test is generally capable of detecting 150-620 ug/L free hemoglobin (or 5 to 20 intact red blood cells per microliter) in urines w/ specific gravity of 1.005 and ascorbic acid concentrates of < 0.3 mmol/L and is less sensitive in urines with higher specific gravity or greater ascorbic acid content. The test is slightly more sensitive to free hemoglobin and myoglobin than to intact red cells. The appearance of green spots on the reacted reagent area indicates the presence of intact red cells in the urine. Certain oxidizing contaminants such as hypochlorite and microbial peroxidase associated with urinary tract infection, may cause false positive results.

V. Urobilinogen (45 seconds) 60 seconds ulit…

Principle: Ehrlich reaction

p-dimethly-amino-benzaldehyde reacts w/ urobilinogen in a strongly acid medium to form a brown orange color.

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Reagent Composition:

Para-dimethylaminobenzaldehyde

This test is based on the Ehrlich reaction in which p-dimethyl-amino-benzaldehyde reacts w/ urobilinogen in a strongly acid medium to form a brown orange color. In healthy population, the normal urine urobilinogen range is 1.6-16 umol/L urobilinogen. The absence of urobilinogen in the specimen cannot be established. The test will react w/ interfering substances known to react w/ Ehrlich’s reagent, such as p-aminosalicyli acid. Drugs containing azo dyes may give a masking golden color. This test is not a reliable method for the detection of prophobilinogen. VI. Specific Gravity (45-60 seconds)

Principle:

This test is based upon the ionic conc of urine. The pKa’s of certain pretreated polyelectrolytes change in relation to the ionic conc of the urine. In the presence of an indicator, colors range from deep blue-green in urine of low ionic conc through green and yellow-green in urines of increasing ionic conc.

A lower S.G. reading

- highly buffered alkaline urines

- urines containing glucose or urea at conc of > 1% Elevated S.G. readings

- presence of moderate 91.0-7.5 g/L) qty of protein. S.G. readings correlate with values obtained by the refractive index method. A lower S.G. reading, relative to the other methods, may be caused by highly buffered alkaline urines and urines containing glucose or urea at conc of > than 1%. Elevated S.G. readings, relative to other methods, may be obtained in the presence of moderate (1.0-7.5 g/L) qty of protein. Random urines may vary in S.G. between 1.003-1.040. This S.G. test permits determination of urine S.G. between 1.000-1.030. For increased accuracy, 0.005 may be added to readings from urines with pH equal or greater than 6.5.

Reagent Composition:

Bromthymol blue, poly (methyl vinyl ether maleic anhydride), sodium hydroxide

VII. pH (may be read immediately – timing not critical)

Principle: Double indicator system

Gives a range of colors from orange through yellow and green to blue and permits differentiation to w/in 0.5 pH units in the range pH 5 to pH 8.5.

Reagent Composition:

methyl red, bromthymol blue, nonreactive ingredients This test is based on the double indicator system principle. Readings are not affected by variations in urinary buffer concentration, but contamination by reagent washed from an overwetted adjacent reagent area may affect results.

VIII. Protein (may be read immediately – timing not critical)

Principle: Protein-error-of-indicators

The presence of protein results in development of a green color

Reagent Composition:

Tetrabromphenol blue, buffer, non-reactive ingredients. Note: contamination of the urine sample w/ chlorhexidine antiseptic will give a false-positive reading for protein.

The test is based on the protein-error-of-indicators principle. The presence of protein results in development of a green color (echo!). The test yields negative results w/ normal urines, so any positive result greater than “Trace” indicates significant proteinuria. Clinical judgment may be used to interpret “Trace” results, w/c may occur w/ urines of high specific gravity w/ non-significant protein-uria. The “trace” result corresponds to 0.05-0.2 g/L albumin, but the test is less sensitive to globulin, Bence-Jones

protein and mucoprotein, so a negative result does not rule out the presence of these proteins. False positive results may occur w/ alkaline, highly buffered urines or in the presence of contaminating quaternary ammonium compounds.

IX. Nitrite (60 seconds)

Principle: Greiss reaction

Conversion of nitrate (derived from diet) to nitrite by the action of principally Gram negative bacteria in the urine

Reagent Composition:

p-arsanilic acid, tetrahydrobenzo(h)-quinolin-3-ol

This test depends upon the conversion of nitrate (derived from diet) to nitrite by the action of principally Gram negative bacteria in the urine. The test is specific for nitrite and will not react w/ any other substance normally excreted in urine. Pink spots or pink edges should not be interpreted as positive result. Any degree of uniform pink color development should be interpreted as a positive nitrite test suggesting the presence of 105 or more organisms per mL, but color development is not proportional to the number of bacteria present. A negative result does not in itself prove that there is not significant bacteriuria. Negative results may occur when UTI are caused by organisms w/c do not contain reductase to convert nitrate to nitrite; when urine has not been retained in the bladder long enough (4 hrs or more) for reduction of nitrate to occur; or when dietary nitrate is absent, even if organisms containing reductase are present and bladder incubation is ample. Sensitivity of the nitrite test is reduced for urines w/ high specific gravity. Ascorbic acid conc of 1.42 mmol/L or greater may cause false negative results w/ specimens containing small amts of nitrite ion (13 umol/L or less).

X. Leukocytes (2 minutes)

Principle:

Granulocytic leukocytes contain esterases that catalyze the hydrolysis of the derivatized pyrrole amino acid ester to liberate 3-hydroxy-5-phenyl pyrrole. This pyrrole then reacts w/ a diazonium salt to produce a purple product.

Reagent Composition:

Derivatized pyrrole amino acid ester, diazonium salt

MICROSCOPIC ANALYSIS OF URINE

1. Fresh or adequately preserved urine specimen (approximately 10-15 m) is placed preferably on a conical tube and centrifuged for 5 minutes.

2. Decant supernatant and the sediment is resuspended with the remaining urine in the tube (usually 0.5 mL or 1.0 mL)

3. Using a pipet, place a drop of resuspended sediment on a clean slide and cover with a cover slip

4. Examine the sediment first using the LPO

5. Shift to HPO to identify specific types of cells, casts, bacteria and crystals

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Manner of Reporting: Elements Report As

RBCs and WBCs Average of RBC or WBC seen in 10 high power fields (HPF)

Casts Average casts seen per cover slip ( __/cs) Epithelial cells Report as:

few, occasional, moderate, many or +,++,+++,++++ Crystals Other elements (Bacteria, yeasts) Report as:

few, occasional, moderate, many or +,++,+++,++++

Read under LPO Casts Hyalin Granular Waxy Pus cell RBC Cells Squamous cells Renal cells Mucus threads Crystals Amorphous urates Uric acid Calcium oxalate Amorphous PO4 Triple PO4 Report As

# of casts seen/ cover slip

Few, occasional, moderate or many OR

+, ++, +++, ++++

Few, occasional, moderate or many OR

+, ++, +++, ++++

Read under HPO RBC

WBC Average of RBC/WBC seen in 10 fields/HPF Yeast cells

Bacteria Few, occasional, moderate or many OR +, ++, +++, ++++

ROUTINE FECALYSIS

There is no substitute for careful visual inspection of the patient’s stool by the physician. Patient’s descriptions of their feces are subjective and inaccurate.

Routine Fecal Analysis:

Laboratory examination of feces is most commonly employed for: 1. Detection of gastrointestinal bleeding

2. Confirmation of steatorrhea

3. Detection and identification of parasites Specimen Collection:

A well-ringed bed pan is a convenient collecting vessel. A cleaned, rinsed and boiled glass jar provides a satisfactory alternative. Female patients must be warned not to contaminate the specimen w/ urine since urine has harmful effects on protozoa. Patients must also be instructed not to overfill and contaminate the container.

Stool collected w/in 72 hrs after barium enema or swallow is unsatisfactory. Similarly, stool from patient taking mineral oil is undesirable.

Detection of Fecal Occult Blood Using Hematest Reagent Tablets Principle:

The test is based on the peroxidase-like activity of hemoglobin in catalyzing the oxidation by peroxide of a chromogen, tetramethylbenzidine, to form a blue color.

Reagent:

Tetramethylbenzidine, strontium peroxide Specimen Collection and Preparation:

Collect fecal specimen uncontaminated w/ urine and test as soon as possible. When testing collected specimens, it is recommended that several segments or portions of the stool be tested; blood from colo-rectal bleeding may be predominantly on the outer surface of the formed stool, and blood from upper portions of the GI tract is not always uniformly dispersed throughout the specimen. A meat-free diet prior to specimen collection is desirable to minimize false positive reactions that can result from ingestion of raw or rare meat. Intake of Vit. C should be restricted because ingestion of large quantities may result in false negative results.

Procedure:

1. Place HEMATEST filter paper on a clean glass or

porcelain plate. Apply a small portion of a collected stool specimen to the HEMATEST filter paper w/ an applicator stick. The smear should be a thin, narrow streak. Do not use an emulsion or suspension.

2. place the tablet on the specimen so that part of the tablet is directly touching the clean filter paper

3. Place one drop of distilled water on the tablet. Allow 5-10 secs for the water to penetrate the tablet. Then add a second drop so that the water runs down the side of the tablet onto the specimen filter paper. If necessary, gently tap side of plate once or twice to dislodge water from top of tablet.

4. For up to two minutes, observe filter paper for appearance of any trace of blue color surrounding the tablet. Ignore color that develops on or directly under the tablet itself and any color appearing on the filter paper after the 2-minute time.

Results:

A positive rxn is indicated by the appearance of any trace of blue color on the filter paper around the tablet w/in the 2-minute reading time. Ignore any color that develops on or directly under the tablet itself and any color appearing on the filter paper after the two-minute time.

Neutral Fat Estimation (Sudan III Staining) Procedure:

1. mix stool specimen thoroughly w/ an applicator stick 2. place a small amt (2-3 mm) on a clean glass slide.

Emulsify w/ a drop of ethyl alcohol. 3. add 2 drops of sudan stain

4.

mix and apply cover slip

5. let stand for 5 mins and examine microscopically Interpretation:

Presence of neutral fat droplets is indicated by yellow-orange to red globules which tend to collect at the edges of the cover slip. In normal stools, there are less than 50 fat globules/highly power field by microscopic examination.

Fecal Trypsin Determination Principle:

x-ray film strips are immersed in diluted fecal emulsions. Proteolytic activity (of trypsin and chymotrypsin) causes digestion of the gelatin emulsion on x-ray film

Procedure:

1. mix duodenal content or a pea-sized fecal material in a test tube w/ 2% sodium carbonate (about 0.5 ml)

2.

place a small amount of gelatin square (xray film) into

the mixture and incubate at 37oC Interpretation:

Complete digestion of gelatin is recognized by complete clearing of x-ray film (the blue tint of the film base should be apparent). Partial digestion is indicated by mixed clear and cloudy film. When there is no digestion, the gelatin remains intact.

-fin-arrrrgggggghhhhhh!!!!!!

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Urinalysis Result Form

Physical Characteristics Microscopic Findings

Color: reddish Casts:

Transparency: turbid Hylaine

pH: 6.0 Granular

Specific Gravity: 1.026 Waxy Pus Cell RBC

Chemical Test Cells:

Glucose: negative RBC: 50-60 / HPF Bilirubin: negative Pus Cell: 1-30 / HPF Ketone: negative Yeast Cell

Blood: ++ Squamous Cells: few

Protein: ++ Renal Cells

Urobilinogen: negative Bacteria Nitrite: negative Mucus Threads Leucocyte esterase: negative Crystals:

Amorphous urates: few Uric Acid: ++++ Calcium Oxalate Amorophous PO4 Triple PO4 Diagnosis: Ureterolithiasis (Uric Acid), Right

Urinalysis Result Form

Physical Characteristics Microscopic Findings Color: dark yellow Casts:

Transparency: turbid Hylaine: 20 – 25 /cs

pH: 6.0 Granular: 15-20 / cs

Specific Gravity: 1.030 Waxy Pus Cell RBC > 50 / cs

Chemical Test Cells:

Glucose: negative RBC: 50-60 / HPF Bilirubin: negative Pus Cell: 2-4 / HPF Ketone: negative Yeast Cell

Blood: + Squamous Cells: few

Protein: +++ Renal Cells: few

Urobilinogen: negative Bacteria Nitrite: negative Mucus Threads Leucocyte esterase: trace Crystals:

Amorphous urates: few Uric Acid:

Calcium Oxalate Amorophous PO4 Triple PO4 Diagnosis: Acute Glomerulonephritis

Urinalysis Result Form

Physical Characteristics Microscopic Findings

Color: yellow Casts:

Transparency: turbid Hylaine: 2-4 / cs

pH: 5.0 Granular: 4-6 / cs

Specific Gravity: 1.035 Waxy

Pus Cell: > 50 / cs RBC

Chemical Test Cells:

Glucose: ++++ RBC: 2-4 / HPF Bilirubin: negative Pus Cell: 50-60 / HPF Ketone: negative Yeast Cell: few Blood: negative Squamous Cells: few

Protein: ++ Renal Cells: +++

Urobilinogen: negative Bacteria: +++

Nitrite: ++ Mucus Threads: +

Leucocyte esterase: ++ Crystals:

Amorphous urates: few Uric Acid

Calcium Oxalate Amorophous PO4 Triple PO4

Diagnosis: Acute Pyelonephritis and Diabetes Mellitus

Urinalysis Result Form

Physical Characteristics Microscopic Findings

Color: reddish Casts:

Transparency: turbid Hylaine: 3 / cs

pH: 7.0 Granular: 12 / cs

Specific Gravity: 1.030 Waxy Pus Cell RBC

Chemical Test Cells:

Glucose: negative RBC: 0-2 / HPF Bilirubin: negative Pus Cell: 1-3 / HPF Ketone: negative Yeast Cell

Blood: +++ Squamous Cells: few

Protein: ++ Renal Cells

Urobilinogen: negative Bacteria Nitrite: negative Mucus Threads Leucocyte esterase: negative Crystals:

Amorphous urates: few Uric Acid:

Calcium Oxalate Amorophous PO4 Triple PO4

Diagnosis: Acute Immune-mediated Blood Transfusion Reaction

Urinalysis Result Form

Physical Characteristics Microscopic Findings Color: yellow brown Casts:

Transparency: turbid Hylaine

pH: 6.0 Granular

Specific Gravity: 1.030 Waxy Pus Cell RBC

Chemical Test Cells:

Glucose: negative RBC: 0-1 / HPF Bilirubin: negative Pus Cell: 0-2 / HPF Ketone: negative Yeast Cell

Blood: negative Squamous Cells: few Protein: trace Renal Cells

Urobilinogen: negative Bacteria

Nitrite: negative Mucus Threads: few Leucocyte esterase: negative Crystals:

Amorphous urates: few Uric Acid:

Calcium Oxalate Amorophous PO4 Triple PO4 Diagnosis: Cholelithiaasis (obstructive jaundice)

Urinalysis Result Form

Physical Characteristics Microscopic Findings Color: yellow Casts:

Transparency: turbid Hylaine: 0-1 / cs

pH: 6.0 Granular: 0-1 / cs

Specific Gravity: 1.028 Waxy Pus Cell RBC

Chemical Test Cells:

Glucose: negative RBC: 0-3 / HPF Bilirubin: negative Pus Cell: 50-60 / HPF Ketone: negative Yeast Cell

Blood: negative Squamous Cells: few Protein: trace Renal Cells

Urobilinogen: negative Bacteria: ++

Nitrite: ++ Mucus Threads: few

Leucocyte esterase: ++ Crystals:

Amorphous urates: + Uric Acid:

Calcium Oxalate Amorophous PO4 Triple PO4 Diagnosis: Urinary Tract Infection (E. Coli)

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Figure

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References

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