PART 2: Routine examination of urine (week 4) Flashcards
- The ability of proteins to alter the color
reaction without altering the pH - The test area is buffered to a constant low so that the changes in color will reflect the presence and concentration of proteins.
- Test area is more sensitive to albumin than globulin, Bence-Jones protein, or mucoprotein
- Indicator – tetrabromphenol blue: (-) yellow; (+) green then blue
Other indicators used:
- Tetrachlorophenol
- Tetrabromosulfonaphthaelin
Reagent Strip Method
contains more amino groups
to accept H+ than other proteins
Albumin
Reagent Strip Method other indicators used:
- Tetrachlorophenol
- Tetrabromosulfonaphthaelin
1) Highly buffered alkaline urine
2) High specific gravity
3) Quaternary ammonium compounds
False positive
a) Proteins other than albumin
b) Microalbuminuria
False negative
Principle: Precipitation of
protein by heat and coagulation
by chemical agents
Note: Persistent cloudiness indicates protein.
HEAT AND ACETIC ACID
(+)result: precipitation
This is a cold precipitation test because it
doesn’t require heating
3% SULFOSALICYLIC ACID TEST (EXTON’S)
White opaque ring at the zone of contact
- Heller’s ring or heller’s heat or nitric acid test
Light cloudiness to a heavy flocculation
Picric acid test
White ring at the point of contact
Robert’s ring test
Highly alkaline urine in Acid precipitation
tests
False negative
Highly alkaline urine in Rgt strip test
False positive
Quaternary ammonium compounds in Rgt strip
False positive
Radiographic contrast media in rgt strip
No effect
Radiographic contrast media heat and acetic acid test
False positive
SSA +TCA ( nephelometer or photometer)
Precipitation
TCA- Biuret
Colorimetric
- Turbidity methods
- Turbidity is measured by photometer and spectrophotometer
TCA ( Trichloraceteic Acid )
SSA (Sulfosalicylic acid)
Dye: Coomasie blue, Ponceau S, Benzethonium chloride, Pyrogallol red molybdate
Dye binding colorimetric methods
- Associated with multiple myeloma,
macroglobulinemia and malignant lymphomas - Suspected if the specimen is turbid between 40oC - 60oC and is clear at 100oC
BENCE JONES PROTEINURIA
1) Heat and HAc Acid test
2) Bradshaw
3) Toluene Sulfonic Acid (TSA) Test
4) Electrophoresis – indicated by single
sharp peak in globulin region; best
method
5) Immunofixation electrophoresis (IFE)
Methods for Bence Jones Protein
indicated by single sharp peak in globulin region; best method
Electrophoresis
at temperatures between
40oC and 60oC
Coagulates
when the temperature
reaches 100oC. (dissolved when boiled)
Dissolves
Unique characteristic of Bence Jones
Coagulates and Dissolves
malignant disorder that
results in infiltration of bone marrow by
plasma cells
Multiple myeloma
Predictor of clinical nephropathy in insulin
dependent diabetes mellitus (diabetic
nephropathy
- Hypertension and cardiovascular risk
- Specimen: random or first morning
urine
MICROALBUMINURIA
Albumin Excretion rate
(AER) is______
20 to 200 µg/min
Principle: IMMUNOLOGIC TEST (Enzyme
Immunoassay)
- Strip contains gold labeled antihuman albumin antibody - enzyme conjugate
- Albumin binds w/antibody & move up the strip
- React with the enzyme substrate
- Sensitivity = 0-10mg/dl
Dip strip into the urine for 5 seconds
- Stand for 1 min
- (-) white (+) pink-red
MICRAL TEST
Principle:Immunologic (immunochromatography)
1) Strip in urine for 3 min - albumin in urine binds with blue latex particles coated with antihuman albumin antibody
2) Bound & unbound particle migrate up the strip unbound particles do not migrate as far as the bound particles
IMMUNODIP
1) Dye binding method
2) Reported in albumin:creatinine ratio
CLINITEK MICROALBUMIN
a) Estimates 24 hr microalbumin excretion
b) Corrects for for overhydration or
dehydration in a randspecimen
c) Creatinine – produced & excreted at a
constant rate
Albumin: Creatinine ratio
produced & excreted at a
constant rate
Creatinine
Principle: Binding of albumin to
sulphonaphthalein dye
- Dye: diiodo-dihydroxy-dinitrophenyltetrabromosulphonaphthalein (DIDNTB)
- Specific & sensitive to albumin
(sensitivity: 8 and 15 mg/dL or 80 to 50
mg/dL)
ALBUMIN
Principle: seudo-peroxidase activity of
copper – creatinine complexes
◦ DBDH-diisopropylbenzenedihydroperoxide
(acts as a peroxide )
◦ TMB- tetramethylbenzidine (chomogen)
CREATININE
DBDH
(acts as a peroxide )
diisopropylbenzenedihydroperoxide
TMB (chomogen)
tetramethylbenzidine
Reagent strip that measures protein-high,
protein-low, creatinine, specific gravity, pH, glucose, ketones, bilirubin, blood, nitrite, leukocytes
MULTISTIX PRO
Albumin sensitivity in multistix pro
10 to 150 mg/L
Creatinine sensitivity in multistix pro
10 to 300 mg/dL
- Leukocytes
- Nitrite
- UROBILINOGEN
- Protein
- pH
- Blood
- Sp Gr
- Ketone
- Bilirubin
- Glucose
MULTISTIX
- Glucose
- PROTEIN HIGH
- PROTEIN LOW
- Ketone
- Sp gravity
- Creatinine
- pH
- Blood
- Nitrite
- Leukocytes
Multistix pro