Aubf Flashcards
Bence jones protein is often detected in urine of patients with:
Multiple myeloma
Bone tumor
Chronic leukemia
Hyperthyroidism
Amyloidosis
Fibrotic disease
Adenocarcinoma of stomach and prostatitis
Precipitation of BJP depends on:
Urine ph (4.6- 5.4)
Ionic strength of the solution
Electrolyte composition
Protein concentration
What are the tests for BJP
SSA
HEAT AND ACETIC ACID (HARRISON TEST)
BRADSHAW TEST
PUTNAM TEST
TOLUENE SULFONIC ACID
OSGOOD-HASKIN TEST
JACOBSON AND MILNER TEST
Proteins denature upon exposure to extreme ph or temperature, causing decrease in solubility, thus precipitation
Sulfosalicylic acid precipitation test
This is performed when SSA result is positive
Heat and acetic acid test (harrison test)
3 tubes are prepared in heat and acetic acid test, what are placed in one of each?
5ml of urine dispensed in each tube
One drop of 33% acetic acid to the first tube
Two drops on the second tube
No addition on the third tube
This is performed when heller’s ring test or SSA is positive
Bradshaw test
This uses 2M acetate buffer ( sodium acetate trihydrate and glacial acetic acid)
Putnam test
It precipitates BJP but does not precipitate albumin. It unmasks globulin only if present in high concentrations
TSA (Toluene Sulfonic Acid)
This uses 50% acetic acid and saturated aqueous solution of NaCl
Osgood-haskin test
This test requires a fresh urine to prevent the conversion of proteins to pseudo-BJP or decomposition of BJP
Jacobson and Milner Test
Concentrated nitric acid is overlaid with urine snd white ring is produced in contact (jacobson and milners test). This may indicate the presence of:
Albumin
Proteoses
Resinosis drugs
BJP
Rings 1-2mm above contact zone are due to:
Water
Urea
Bile
Jacobson and milners test:
If the result is positive, procedure must be repeated by using?
Concentrated HCL then 25% acetic acid. If white ring is still present after warming at 70C, BJP is present
Four commonly used brands of commercial reagent strips:
Multistix
Chemstrips
Vchemstrips
Aution stick
4 reagent strip parameter:
Glucose (30s)
Specific gravity (45s)
Ph (60s)
Protein (60s)
Reagent strips (ph):
Erroneous results can occur from?
Improper storage of the specimen (falsely increased)
Contamination of the specimen container (falsely increased)
Improper reagent strip technique (falsely decreased ph)
Runover of urine (falsely acidic in an alkaline urine)
Tests for urine ph
Reagent strips
Ph test paper
Ph meter
It consists of a silver-silver chloride indicator electrode with a ph sensitive glass membrane connected by a salt bridge to a reference electrode (calomel electrode)
Ph meter
What proteins are usually not detected in the reagent strip for albumin?
Globulin
Myoglobin
Hemoglobin
Immunoglobulin light chains
Mucoproteins
Reagent strip in albumin:
False positive reactions are observed in:
Extremely alkaline (ph > 9) or highly buffered
Highly colored substances such as phenazopyridine drugs and beets
Reagent strip (albumin) sensitivity
Chemstrip 6.0mg/dl
Multistix 15 to 30 mg/dl
Vchem at 15mg/dl
Sensitive albumin tests
Immunodip test
Micral test strips
Clinitek microalbumin test strip
Multistix pro test strip
Micral test method
Falsw positive results are caused by;
Ocytetracyline
Strong oxidizing agents
Wualitative tests for protein
Heat and acetic acid test
Purdy’s modification of heat and acetic acid test
Heat coagulation method by henry
Sulfosalicylic acid precipitation test
Sulfosalicylic acid precipitation tests
False positive:
False negativen:
False positive:
Penicilin
Contrast media (crystalline precipitate)
False negative:
Highly alkaline or buffered urine
This involves 3% picric acid to precipitate proteins
Picric acid test
This uses 50% acetic acid and saturated sodium chloride
Osgood haskin test
This uses glacial acetic acid and 5-10% potassium ferrocyanide
Positive result: faint cloudiness appears at once
Potassium ferrocyanide test
Ring or contact tests:
Heller’s ring test
Robert’s ring test
Spiegler’s test (jolles modification)
Spiegler’s reagent consists of:
Mercuric chloride
Succinic scid
Sodium chloride
Distilled water
Hellers ring test
False positive results occurs from:
Uric acid
Urea
Mucin
Biuret reaction
Violet mixture:
Rose colored:
Violet: albumin and globulin
Rose: albumin and peptone
Esbach’s reagent:
Tsuchiya’s reagent:
1% picric acid
2 citric scid
Distilled water
Tsuchiya’s reagent:
Phosphotungstic acid
95% alcohol
Conc. HCl
Quantitative test for proteins
Esbach’s test
Kingsbury’s - Clark Method (SSA)
Shevky and Stafford Method
Kwilecki’s modification of esbach’s
Modified tsuchiya’s test
Purdy’s method
This involves the use of 50% acetic acid snd 10% potassium ferrocyanide added with 15ml of urine. The amount of precipitate after 10 minutes is measured
Purdy’s method
This uses ethanolic HCl phosphotungstic acid to precipitate proteins at 56C followed by spectrophotometry at 540nm
Modified tsuchiya’s test
Involves heating s mixture of 10% ferric chloride and urine at 72C to allow precipitation
Kwileckis Modification of Esbach’s
This uses Tsuchiya’s reagent to precipitate proteins and the degree of flocculation is quantified
Shevky and Stafford method
