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8/18/2019 dipstik lengkap

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Urinalysis Practicum

Clinical Pathology Dept of Medical Fac

Brawijaya Univ/Saiful Anwar

General Hospital Malang


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Sample Processing

Sentrifuge 1500-

2000 rpm, 5 min

For chemical

examinationFor microscopic (0.5

cc)

10-12 ml

Urine


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Sentrifuge 1500-

2000 rpm, 5 min

For microscopic (0.5 cc)

10-12 ml

urine, mix:

forchemical

dipstick


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Urine Protein (Boiling method)

- : Clear

+1 : slightly cloudy+2 : Cloudy, seed

+3 : Cloudy,

fragmented

+4 : Cloudy,agglutinate

3-5 drops

3-6%

acetic acid

2 mlUrine

Test controle


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Negative + 1 + 2 + 3 + 4

Interpretation


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Glucose urine (reduction test)

• Principle:

Glucose will reduced cupri (Cu3+) to

cupro (Cu

2+

)


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Negative Positive 3+

Benedict method

8 drops

Urine

5 ml

Reagent

Boil


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Interpretation

Negative + 1+ 2 + 3 + 4

Negative (-) : Blue or greeny blue, cloudy

Positive 1+ : yellowish green and cloudy

Positive 2+ : yellow, cloudy

Positive 3+ : orange, cloudy

Positive 4+ : brownish red


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Dipstick & Microscopic

Examination


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The Procedure of Dipstick Exam

1. Test urine as soon as possible after

receipt (use fresh urine)

2. Remove only enough strips forimmediate use; recap tightly.

3. Test a well-mixed, unspun urine sample.

4. Do not touch the test area with fingers.

5. Dip reagent strip into urine briefly – nolonger than 1 second.


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6. Drain excess urine off – run edge ofstrip along rim of tube

7. Touch the edge of the strip to an

absorbent paper

8. Do not lay reagent strip directly onworkbench surface

9. Follow exact timing recommendations

for each chemical test. Usually 60 sec,

no more than 2 min

10. Hold reagent strip close to the color

chart and read under good lighting.


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Urine Multistix – reading dipstick

results manually; colors are matched

to those on the bottle label; timing is

critical for each pad.


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• Polyelectrolyte sensitive to ions in

urine

• If electrolytes level is increased, SG is

increased too

1. Polyelectrolyte: polymethylvinyl ether/maleic acid

2. Indicator: bromthymol blue

Specific gravity


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pH

Manufacturers use a double-indicator system

of methyl red and bromthymol blue.

Methyl red H → Bromthymol blue H

(Red-Orange → Yellow) (Green → Blue)


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Protein

The protein area of the strip contains

either tetrabromphenol blue or 3′ , 3′′ , 5′ ,

5′′ -tetrachlorophenol-3, 4, 5, 6-

tetrabromosulfonphthalein and an acidbuffer to maintain the pH at a constant

level.


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Negative

Positive

Readings are reported in terms of negative,

trace, 1+, 2+, 3+, and 4+; or thesemiquantitative values of 30, 100, 300, or

2000 mg/dL corresponding to each color

change.


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Source of error/interference

False positive False negative

Highly buffered alkaline

urine

Pigmented specimens,

phenazopyridine

Quaternary ammoniumcompounds (detergents)

Antiseptics, chlorhexidine

Loss of buffer from

prolonged exposure of thereagent strip to the specimen

High specific gravity

Proteins other than albumin

Microalbuminuria


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Glucose

glucose oxidase

1. Glucose + O2 (air) → gluconic acid H2O2

peroxidase

2. H2O2 + chromogen → oxidized colored chromogen +

H2O


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Interference

• False-positive:Contamination by oxidizing agents and

detergents

• False-negative:High levels of ascorbic Acid,

High levels of ketones

High specific gravity

Low temperatures

Improperly preserved specimens


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Keton

• Results are reported qualitatively asnegative, trace, small (1+), moderate (2+),

or large (3+), or semiquantitatively as

negative, trace (5 mg/dL), small (15

mg/dL), moderate (40 mg/dL), or large (80to 160 mg/dL).

alkaline

acetoacetate + sodium nitroprusside + (glycine) → (and acetone) purple color


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Blood

hemoglobin

H2O2 + chromogen → oxidized chromogen + H2Operoxidase green-blue color

• False-positive: Bacterial peroxidases,Menstrual contamination

• False-negative: High specific

gravity/crenated cells, Formalin, Captopril,High concentrations of nitrite, Ascorbic

acid 25 mg/dL, Unmixed specimens


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Bilirubin

acid

bilirubin glucuronide + diazonium salt → azodye

• False-positive: Highly pigmented urines,

phenazopyridine, Indican (intestinal

disorders), Metabolites of Lodine

• False-negative: Specimen exposure to light,Ascorbic acid 25 mg/dL, High

concentrations of nitrite


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Urobilinogen

acidurobilinogen + p-dimethylaminobenzaldehyde→ red color

(Ehrlich’s (Ehrlich reagent)

reactive

substances)

• False-positive: Porphobilinogen, Indican, p-

aminosalicylic acid, Sulfonamides, Methyldopa,

Procaine, Chlorpromazine, Highly-pigmentedurine

• False-negative: Old specimens, Preservation in

formalin

k


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Leukocyte esterase

leukocyte esterases

indoxylcarbonic acid ester → indoxyl + acid indoxylacid

+ diazonium salt → purple azodye

• False-positive: Strong oxidizing agents,

Formalin, Highly pigmented urine,

nitrofurantoin

• False-negative: High concentrations of

protein, glucose, oxalic acid, ascorbic acid,gentamicin, cephalosporins, tetracyclines,

inaccurate timing


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Nitrite

acid

para-arsanilic acid or sulfanilamide NO2 → diazonium salt

(nitrite)

aciddiazonium salt tetrahydrobenzoquinolin → pink azodye

P-arsanilic

acid

positive


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2000 rpm/5 min

10 – 12 ml urine

in to centrifuge

tube Pour supernatant, quickly backbut smoothly, stand on rack,sediment ± 0.5-1 ml

Microscopic Urinalysis (Sediment)


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Shake to resuspendsediment, place 1 drop on aslide

Coverslip the drop

• Place on the slide on the microscope.• Reduce your light setting and lower

the condenser approx. halfway down.

• If the light is still too bright, close the

diaphragm slightly.


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• Low power field (10X Obj)

– Cast count no. (range) – Epithelial cells (+/-, types)

– Crystals (+/-, types)

• High power field (45X obj)

– Erythrocytes:

•Count no. range

•Morphology eu/dysmorphic,

crenated – Leukocytes:

•Count no. range


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SedimentErythrocytes : N: 0 – 2/hpf

Dismorfik


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Normal: 0 – 5/hpf

Leukocytes Epithelial cells


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Hyaline cast


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Coarsely granular cast Finely granular


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Epithelial cast


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Leukocyte cast


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Erythrocyte cast


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Casts


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Oval Fat odies

Trichomonas

Waxy cast Rough granular cast


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Crystals

Uric acid

Ca oxalate


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Ca

phosphate

n

TriplePhosphate


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Tyrosine crystal

R l R i


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Result Reporting

• Chemical stick:

- Glucose: ...

- Protein : ...- Keton :… etc.

• Sediment:

– Cast : + / -

• Hyaline : …. - …. / LPF• Erythrocyte : …. - …. / LPF

• Leukocyte : …. - …. / LPF

• Granular : …. - …. / LPF

• Etc . : …. - …. / LPF

– Ery : …. - …. / HPF (eu/dysmorphic)

– Leuko : …. - …. / HPF

– Epithel : + / ++ / +++, Type: ……..

– Crystal : + / ++ / +++, Type: ……..

– Other(s) : ……………………………….…….


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