KLUBSY: CHEMICAL ANALYSIS OF URINE Flashcards
These provide, simple, rapid means for performing medically significant analysis of urine
Reagent strips
This consist of chemical-impregnated absorbent pads attached to a plastic strip.
Reagent strips
Reagent strips consist of what?
consist of chemical-impregnated absorbent pads attached to a plastic strip
Reagent strip consist of a chemical-impregnated absorbent pads attached to a what?
attached to a plastic strip
A color producing chemical reaction takes place when the absorbent pad comes in contact with urine
reagent strip
What is used for reagent strip testing?
a fresh, well-mixed, uncentrifuged specimen
What are the 10 parameters of a rgnt strip?
1 - pH
2- protein
3-glucose
4-ketones
5-blood
6-bilirubin
7-urobilinogen
8-nitrite
9-leukocytes
10-S.G
What is the 11th parameter in a rgnt strip?
REDUCING AGENT VITAMIN C (Ascorbic acid)
How long must you dip the rgnt strip?
briefly; NO LONGER THAN 1 SECOND
T/F: The edge of a rgnt strip should be blotted on a disposable absorbent pad
T
Remedy: Formed elements such as WBC and RBC sinks to the bottom of the specimen and will be undetected
MIX SPECIMEN WELL
Allowing the strip to remain in the urine for an extended period may cause what?
may cause leachinf of rgnts from the pads
How to ensure against run-over in rgnt strip?
BLOT the edge of the strip with ADSORBENT paper and holding the strip HORIZONTALLY FACING DOWNWARD while comparing it with color chart
T/F: Specimens that have been refrigerated must not be returned to room temp prior to rgnt strip testing, as enzymatic rxns on the strip are temperature dependent
F.
MUST BE RETURNED TO ROOM TEMP
At what temperature must rgnt strips be stored?
Stored BELOW 30C (room temp); DO NOT FREEZE
Why must specimens that have been refrigerated be returned to room temperature before chemical testing by rgnt strips?
because the enzyme reactions on the strips perform best at room temperature
How is the QC for reagent strips?
Rgnt strips must be checked with BOTH positive and negative control a minimum of once every 24 hours.
When must rgnt strip be quality checked
minimum of once every 24 hrs
This is NOT recommended as a negative control
Distilled water
T/F: Distilled water is recommended as a negative control
F.
NOT recommended
Why is distilled water NOT recommended as a negative control?
because rgnt strip chemical rxns are designed to perform at IONIC CONCENTRATIONS similar to urine.
These are defined as test using different reagents or methodologies to detect the same substances as detected by the rgnt strips with the same or greater sensitivity or specificity.
Confirmatory tests
These procedures use tablets and liquid chemicals that may be available when questionable results are obtained or highly PIGMENTED specimens are encountered
Non-reagent strip testing
This is important in the identification of urinary crystals and determinastion of unsatisfactory specimens
pH
Important in aid of existence of systemic acid-based balance disorders.
pH
This is controlled primarily by dietary regulation, although medications also may be used
Urinary pH
Urinary pH is controlled primarily by what?
dietary regulation
This can be used for determination of unsatisfactory specimens
Urinary pH
Urinary pH can be used for determination of what?
Unsatisfactory specimens
This may be used for the treatment of urinary tract infections and renal calculi formation and prevention
Urinary pH
What is the normal urine pH?
4.5 - 8
what is the first morning urine pH?
5-6
This indicated a collection of a fresh sample
improperly preserved specimen
What is the pH of an improperly preserved specimen?
> 9 or > 8.5
This can cause alkalization of urine
Presence of detergent in the urine container
Presence of detergent in the urine container can cause what?
alkalization of urine
What is the most affected parameter in an unmixed urine sample?
BLOOD
This caused when the rgnt strip remains in contact with urine for extended or longer period of time (>1 second)
LEACHING
What is the remedy for leaching?
the rgnt strip should NOT be dipped in urine for more than 1 second to prevent leaching
Leaching is caused by what?
when the rgnt strip remains in contact with urine for extended or longer period of time (>1 second)
This is caused when the rgnt strip was NOT BLOTTED or was improperly blotted
Run-over
What is the remedy for a run-over
blotting the edge of the rgnt strip WITH ADSORBENT PAPER
A run-over in a rgnt strip is caused by what?
when the rgnt strip was NOT BLOTTED or was improperly blotted
This will lead to distortion of colors in the rgnt pad
RUN-OVER
Which of the following parameters is most affected when using a refrigerated sample?
GLUCOSE
A substance that abosrbs moist
hygroscopic
For QC of rgnt strip, what is used for negative control?
TAP WATER
When to a reject a sample?
if pH is > 9 or >8.5
*REJECT and recollect
What would be the result if there is detergent left in the container?
FALSE ELEVATION
What is the principle of the rgnt strip?
DOUBLE INDICATOR SYSTEM
What are the 2 indicators in rgnt strip?
- Methyl red (detects acidic urine)
- Bromthymol blue (detects alkaline pH)
What is the product of an acidic urine pH?
FROM RED TO YELLOW
What is the product of an alkaline pH
FROM GREEN TO BLUE
What is the pH of the methyl red indicator?
pH 4.0-6.0
What is the pH of the indicator bromthymol blue?
pH 6.0-9.0
What are the reagents used in the rgnt strip?
METHYL RED AND BROMTHYMOL BLUE
Which is associated with acidic urine?
A. Apple
B. Cranberry
C. Orange
D. Pomelo
CRANBERRY
Assoc. with acidic or alkaline urine: Emphysema
Acidic
Assoc. with acidic or alkaline urine: DM
Acidic
Assoc. with acidic or alkaline urine: Starvation
Acidic
Assoc. with acidic or alkaline urine: Dehydration
Acidic
Assoc. with acidic or alkaline urine: Cranberry juice
Acidic
Assoc. with acidic or alkaline urine: High protein diet
acidic
Assoc. with acidic or alkaline urine: food rich in fats/lipids
acidic
Assoc. with acidic or alkaline urine: E.coli
Acidic
*E.coli is an acid-producing bacteria
Assoc. with acidic or alkaline urine: Mandelamine
acidic
Assoc. with acidic or alkaline urine: Fosfomycintromethamine
acidic
Assoc. with acidic or alkaline urine: renal tubular acidosis
alkaline
Assoc. with acidic or alkaline urine: hyperventilation
alk
Assoc. with acidic or alkaline urine: vomiting
alk
Assoc. with acidic or alkaline urine: vegetarian diet
alk
Assoc. with acidic or alkaline urine: old specimens
alk
Assoc. with acidic or alkaline urine: presence of urease producing bacteria
alk
Assoc. with acidic or alkaline urine: alkaline tide (during and after following meals)
alk
Assoc. with acidic or alkaline urine: Proteus spp
alk
*presence of urease-producing bacteria
Assoc. with acidic or alkaline urine: Pseudomonas spp
alk
*pseudomonas spp is a urease-producing bacteria
Assoc. with acidic or alkaline urine: pomelo
alk
Assoc. with acidic or alkaline urine: lime
alk
Assoc. with acidic or alkaline urine: orange
alk
Assoc. with acidic or alkaline urine: lemons
alk
Assoc. with acidic or alkaline urine: grapefruits
alk
Density of a solution compared with density of similar volume of distilled
water at a similar temperature
S.G
S.G is influenced by what?
by number and size of particles on a solution
Rgnt strip s.g test does not measure the total solute content but only those solute that are what??
solutes that are IONIC
Normal random s.g
1.002 - 1.030
Radiographic contrast dye
S.G = > 1.040
S.G of NOT a urine
S.G = <1.002
What is the principle of S.G?
Change in the pKa of a polyelectrolyte in an alkaline medium
What is the indicator for S.G?
bromthymol blue
in S.G, the more H+ molecules are added to bromthymol blue, the more ??? it becomes
THE MORE ACIDIC it becomes
*since bromthymol blue is alkaline
What is the color change from bromthymol blue (alkaline) to more H+?
Blue ( decreased H+) > Green (++H) > YELLOW (+++H)
This parameter measures IONIC solutes ONLY
S.G
Why must S.G measure only IONIC solutes?
to eliminate the interference by large organic molecules (e.g., urea and glucose, and by radiographic contrast)
What are the rgnts used in S.G?
-Multistix
- Chemstrip
Which reagent in S.G consists of the “poly” and bromthymol blue?
Multistix
Multistix is composed of?
poly (methyl vinyl ether/maleic anhydride), bromthymol blue
Which reagent in S.G consists of ethylene glycol and bromthymol blue?
Chemstrip
Chemstrip is composed of what?
Ethylene glycol diaminoethyl ether tetra acetic acid and bromthymol blue
Sensitivity of S.G parameter?
1.000 to 1.030
In S.G parameter, what S.G is if product is blue?
1.000
in the S.G parameter, what is the S.G if product is YELLOW?
1.030
In the S.G parameter, what color is produced if s.g is 1.000
blue
in the S.G parameter, what color is produced if the s.g is 1.030?
yellow
In pH paramater, what are the sources of error/interferences?
-No known interfering substances
- Run over from adjacent pads
- Old specimens
In the S.G parameter, what are its interferences?
- False positive = High conc. of PROTEIN (100-500 mg/dL), KETOACIDOSIS
- False negative = Highly ALKALINE urines ( > pH 6.5 - add 0.005 SG reading)
What is the result for the interference of high conc. of protein?
False +
what is the result in the presence of the interence ketoacidosis?
false +
what is the result due to the interference of highly alkaline urines?
false (-)
T/F: The higher the concentration of urine, the more hydrogen ions are released, thereby lowering the pH
T
This parameter is most indicative of renal disease
PROTEIN
T/F: Normal urine contains very little protein
T
How much protein is excreted per 24 hours?
less than 10 mg/dL or 100 mg per 24 hours
Normal urine contains how much protein?
<150/24 hours
This is the major protein found in normal urine due to its low molecular weight
ALBUMIN
What other proteins are normally found in the urine?
- Serum and tubular microglobulins
- Tamm-horsfall protein (uromodulin)
- protein from prostatic and vaginal secretion
This produces a white foam in urine
protein
Protein produces what colored foam in urine?
white foam
What is the value of CLINICAL PROTEINURIA?
greater than or equal to 30 mg/dL or greater or equal to 300 mg/L
This is caused by condition that affect the plasma prior to its’ reaching the kidney
Pre-renal or overflow proteinuria
4 factors in pre-renal or overflow proteinuria
A - Hemoglobin
B- Myoglobin
C-Acute phase reactants
D-Bence Jones protein
this is associated with intravascular hemolysis
Hemoglobin
this is associated with muscle injury
myoglobin
this is associated with inflammation and infections
acute phase reactants
this protein is associated with multiple myeloma
Bence Jones protein
An immunoglobulin light chains (kappa and lambda) found in cases of multiple myeloma
Bence Jones protein
What are the immunoglobulin light chains in Bence Jones protein?
kappa and lambda light chains
Confirmatory test for Bence Jones protein
Serum electrophoresis (Immuno-electrophoresis/Immunofixation)
Heat precipitation test in urine: coagulates/precipitates/insoluble at what???
40-60C
heat precipitation test in urine: dissolves/SOLUBLE at ???
80-100C
*100 is the most preferred
This is known as the TRUE renal disease
Renal proteinuria
What happens in glomerular proteinuria?
When the glomerular membrane is damaged,
- selective filtration is impaired
-increased amounts of serum protein
- red and white blood cells pass through the membrane and are excreted in the urine
How much amount of protein appears in the urine following glomerular damage?
ranges from slightly above noram to 4 g/day
Examples of GLOMERULAR PROTEINURIA
1-Diabetic Nephropathy/Kimmelstiel-Wilson’s Disease
2-Amyloidosis
3-Immune complexes found in SLE and Streptococcal glomerulonephritis
4-Toxic substances
5-Pre-eclampsia and eclampsia
6-orthostatic/cadet/postural proteinuria
2 types of renal proteinuria
1-glomerular proteinuria
2-tubular proteinuria
This is when normally filtered albumin can no longer be reabsorbed
Tubular proteinuria
examples of TUBULAR PROTEINURIA
-Fanconi syndrome
-toxic agents/heavy metals (such as cadmium dust)
- several viral infections
This is assoc. in lower UTI/inflammations, injury/trauma, menstrual contamination, prostatic fluid/spermatozoa, and vaginal secretion
Post-renal proteinuria
Type of proteinuria: Lower UTI/inflammations
Post renal
Type of proteinuria: Injury/trauma
post renal
Type of proteinuria: menstrual contamination
post renal
Type of proteinuria: prostatic fluid
post renal
Type of proteinuria: spermatozoa
post renal
Type of proteinuria: vaginal secretion
post renal
Type of proteinuria: fanconi’s syndrome
renal
Type of proteinuria: toxic agents/heavy metals
renal
Type of proteinuria: cadmium dust
renal
Type of proteinuria: severe viral infections
renal
Type of proteinuria: Diabetic nephropathy
renal
Type of proteinuria: Kimmelstiel-Wilson’s disease
renal
Type of proteinuria: Amyloidosis
Renal
Type of proteinuria: Immune complexes found in SLE
Renal
Type of proteinuria: immune complexes found in streptococcal glomerulonephritis
Renal
Type of proteinuria: pre-eclampsia
renal
Type of proteinuria: eclampsia
renal
Type of proteinuria: orthostatic/Cadet/Postural proteinuria
Renal
Type of proteinuria: Bence Jones protein
Pre-renal
This is a.k.a overflow proteinuria
pre-renal
This is usually transient and can be produced by condition such as strenuous exercise, high fever, dehydration, and exposure to cold.
Benign proteinuria
Normal Albumin Excretion Rate (AER)
0-20 ug/min
microalbuminuria value in AER?
20-200 ug/min or 30-300mg/24 hrs
What pH environment is produced in HIGH S.G?
ACIDIC environment
what pH environment is produced in LOW S.G?
Alkaline environment
What protein is seen in the urine that is unique to the kidney
Uromodulin
this protein is produced by the RTE cells
Uromodulin
Uromodulin is a.k.a
Tamm-Horsfall protein
BJP was first described in a patient admitted to where?
St. George’s Hospital in London
BJP was first described in a patient in London under the care of ???
Drs. Watson and MacIntyre (1845)
BJP was discovered by?
Dr. Henry Bence Jones
What was found in the urine by Dr. Henry Bence Jones?
A substance that was precipitated by the addition of nitric acid
What did Dr. Jones call the substance he found in the urine?
Hydrated deutoxide of albumen
BJP is a.k.a
Free light chains
BJP are present in the serum as what?
tetramers
Laboratory analysis of patient urine can identify Bence Jones proteins at a threshold of what?
threshold of 10mg/L
What is the appearance of BJP found in the urine?
-monomers
-dimers
-or high molecular weight polymers
What type is the preferred urine specimen for BJP?
24HRS URINE SAMPLE
What type is the ALTERNATIVE urine specimen in BJP?
MORNING URINE SAMPLE/first or second morning
This type of urine specimen is for the screening of BJP?
Morning uring sample (either first or second morning)
Lower UTI is a.k.a
Cytitis or Urethritis
Urethritis is a.k.a
LOWER UTI
Upper UTI is a.k.a?
pyelonephritis
Pyelonephritis is a.k.a
Upper UTI
Type of proteinuria: Upper UTI
Renal
What organ is affected in renal proteinuria?
KIDNEYS
What organ is affected in POST renal proteinuria?
Bladder or the urethra
This is associated with renal failure in persons with Type I and II DM
Diabetic Nephropathy / Kimmelstiel-Wilson’s disease
Diabetic Nephropathy: Decreased or increased GFR?
DECREASED
What is the indicator for Diabetic nephropathy/Kimmelstiel-Wilson’s disease?
MICROALBUMINURIA
This is due to the presence of albumin in urine above the normal level but below the detectable range of conventional urine dipstick methods
Microalbuminuria
The presence of this is also associated with an increased risk of cardiovascular disease
Microalbuminuria
The presence of microalbuminuria is also associated with an increased risk of what?
Cardiovascular disease
T/F: All glomerular disorders are characterized by PROTEINURIA
T
What is the minimum sensitivity of a rgnt strip for albumin?
10 mg/dL
*2nd answer = 15 mg/dL
What is the major contributor in orthostatic proteinuria?
POSTURE
What are the 2 types of urine specimen for orthostatic proteinuria?
- 1st morning
- 2 hours after standing
This is the oldest for microalbuminuria
AER - Albumin Excretion Rate
What are the 2 types of immunologic tests for testing for microalbuminuria
- micral-test
-ImmunoDip
What is the principle for micral-test?
Enzyme immunoassay
Sensitivity value of a micral-test?
0 to 10 mg/dL
Reagents in Micral-Test
- Gold-labeled antibody
- B-galactosidase
-Chlorophenol red galactoside
What is the principle for ImmunoDip?
Immunochromographics
What is the sensitivity value for ImmunoDip?
1.2 - 8.0 mg/dL
What are the reagents for an ImmunoDip?
-Antibody-coated blue latex particles
What are the interferences in an ImmunoDip?
False-neg: Dilute urine
Strips are dipped into the urine up to a level marked on the strip and helpd for 5 seconds
Micral-Test
Strips are individually packaged in specially designed containers. The container is placed in the urine specimen for 3 minutes
ImmunoDip
In the Micral-Test, strips are dipped into the urine up to a level marked on the strip and help for how long?
held for 5 seconds
In the ImmunoDip, strips are individually packaged in specially designed containers. the container is placed in the urine specimen for how long?
3 minutes
Reading time for Micral-test?
1 minute
What color is produced in a negative micral-test?
WHITE COLOR
What color is produced in a positive micral-test?
RED
*2nd answer: PINK or BLUE
*3rd answer: BLUE
What is the appearance of a negative immunodip result?
Darker bottom band
What is the appearance of a borderline ImmunoDip result?
Equal band colors
What is the appearance of a positive Immunodip result?
Darker TOP band
What is the amount sensitivty of a negative ImmunoDip?
<1.2
What is the amount sensitivity of a borderline ImmunoDip?
1.2 to 1.8
What is the amount sensitivity of a positive ImmunoDip?
2 to 8
What are the interferences in the Micral-Test?
- False positive: Strong oxidixing agent (soap)
- False negative: Dilute urine
What is chromogen in Micral-test?
Chlorophenol red galactoside
What reagent has the HIGHEST sensitivity and specificity to albumin?
a-DIDNTB
b-DBDH
c-TMB
d-Tetrabromphenol blue
DIDNTB
What is the abnormal A:C ratio
30 to 300 mg/g or 3.4 to 33.9 mg/mmol
What are the kinds of A:C ratio rgnt strips?
-Albumin strip
-creatinine strip
Albumin reagent strips use what dye?
bis (3’,3”-diiodo-4’, 4”-dihydroxy-5’,5”-dinitrophenyl)-3,4,5,6-tetrabromo sulphonphthalein (DIDNTB)
This has a higher sensitivity and specificity for albumin
DIDNTB
DIDNTB strips can measure albumin between what range?
between 8 and 15 mg/dL (80 to 150 mg/L) WITHOUT inclusion of other proteins
What is the color range in an albumin strip?
Pale green to Aqua blue
What are interferences in an albumin strip?
-HIGHLY BUFFERED ALKALINE URINE can be controlled using paper treated with bis-(heptapropylene glycol) carbonate
-FALSELY ELEVATED results can be caused by visibly BLOODY URINE, and abnormally COLORED URINES
The addition of this decreases the nonspecific binding of polyamino acids to the albumin pad
Addition of polymethyl vinyl ether
What is the principle of a creatinine strip?
Pseudoperoxidase acitivity of copper-creatinine complexes
What are the reagents of a creatinine strip?
-Copper sulfate
-3,3’,5,5’-tetramethylbenzidine (TMB)
-Diisopropyl benzene dihydroperoxide (DBDH)
What is the color range of a creatinine strip?
Orange (negative) to green to blue
T/F: NO creatinine reading are considered abnormal, as creatinine is normally present in concentrations of 10 to 200 mg/dL
F.
10 to 300 mg/dL
The purpose of this is to correlate the albumin concentration to the urine concentration
the purpose of creatinine measurement
What are the interferences in a creatinine strip?
-Falsely increased: visibly bloody urine, presence of the gastric acid-reducing medication cimetidine (Tagamet), and abnormally colored urines
The only protein that can be detected by a rgnt strip
ALBUMIN
What is the principle for the rgnt strip reaction for protein?
Protein (Sorensen’s) error of indicator
Sorensen’s test is more sensitive to albumin because?
because albumin contains more amino groups to accept the hydrogen ions than other proteins
T/F: The pH of the medium in Sorensen’s test remains constant
T
The pH of the medium in the Sorensen’s test is:
pH of 3 buffered with citrate
T/F: Reagent strip is sensitive to albumin ONLY
T
In the Sorensen’s test, what is the color for negative?
yellow
*Indicators appear YELLOW in the ABSENCE of protein; however, as the protein concentration increases, the color progresses through various shades of green and finally to blue
In the Sorensen’s test, what is the color for positive?
Blue-Green
*Indicators appear YELLOW in the ABSENCE of protein; however, as the protein concentration increases, the color progresses through various shades of green and finally to blue
What are the reagents of a Sorensen’s test?
-Multistix
-Chemstrip
What is the indicator utilized in Multistix?
Tetrabromphenol blue
What is the indicator utilized in Chemstrip?
Tetrachlorophenoltetrabromosulfonphthalein
In the grading for Sorensen’s test, what is the grading for an albumin quantitiy for <30 mg/dL
Trace
In the grading for Sorensen’s test, what is the grading for an albumin quantitiy for 30 mg/dL
1+
In the grading for Sorensen’s test, what is the grading for an albumin quantitiy for 100 mg/dL
2+
In the grading for Sorensen’s test, what is the grading for an albumin quantitiy for 300 mg/dL
3+
In the grading for Sorensen’s test, what is the grading for an albumin quantitiy for 2000 mg/dL
4+
False pos/False neg interference in Sorensen’s test: microalbuminuria
False neg
*Remember that only microalbuminuria and proteins other than albumin are the only ones that result in a FALSE NEGATIVE interference
False pos/False neg interference in Sorensen’s test: Proteins other than albumin
false neg
*Remember that only microalbuminuria and proteins other than albumin are the only ones that result in a FALSE NEGATIVE interference
False pos/False neg interference in Sorensen’s test: Highly buffered interference alkaline urine
false pos
False pos/False neg interference in Sorensen’s test: Pigmented specimens, phenazopyridine
false pos
*Remember that only microalbuminuria and proteins other than albumin are the only ones that result in a FALSE NEGATIVE interference
False pos/False neg interference in Sorensen’s test: Quaternary ammonium compounds (detergents)
False pos
*Remember that only microalbuminuria and proteins other than albumin are the only ones that result in a FALSE NEGATIVE interference
False pos/False neg interference in Sorensen’s test: Antiseptics, chlorhexidine
false pos
*Remember that only microalbuminuria and proteins other than albumin are the only ones that result in a FALSE NEGATIVE interference
False pos/False neg interference in Sorensen’s test: Loss of buffer from prolonged exposure of the strip to the specimen reagent
False pos
*Remember that only microalbuminuria and proteins other than albumin are the only ones that result in a FALSE NEGATIVE interference
False pos/False neg interference in Sorensen’s test: High specific gravity
false pos
*Remember that only microalbuminuria and proteins other than albumin are the only ones that result in a FALSE NEGATIVE interference
T/F: S.G of the urine should be considered in evaluating urine protein because a trace protein in a dilute specimen is MORE SIGNIFICANT than in a concentrated specimen
T
What does the SSA precipitation test mean?
Sulfosalicylic acid precipitation test
This is a cold precipitation test that reacts equally with all forms of protein
SSA PRECIPITATION TEST
In the SSA test, the final solution concentration (urine + reagent) is what?
0.015 g of SSA per milliliter of total solution
SSA grading: No increase turbidity
Negative
SSA grading: Noticeable turbidity
Trace
SSA grading: Distinct turbidity, with no granulation
1+
SSA grading: Turbidity with granulation, no flocculation
2+
SSA grading: Turbidity with granulation, and flocculation
3+
SSA grading: Clumps of protein
4+
SSA interferences: Highly alkaline urine
False decrease/false neg
*Remember: In the SSA test, only HIGHLY ALKALINE URINE and QUATERNARY AMMONIUM COMPOUNDS (e.g detergents and soap) result in FALSE DEC/FALSE NEG
SSA interferences: Quaternary ammonium compounds (e.g detergents and soap)
false decrase/false neg
*Remember: In the SSA test, only HIGHLY ALKALINE URINE and QUATERNARY AMMONIUM COMPOUNDS (e.g detergents and soap) result in FALSE DEC/FALSE NEG
SSA interferences: Radiographic contrast dye/x-ray film
False increase/false pos
*Remember: In the SSA test, only HIGHLY ALKALINE URINE and QUATERNARY AMMONIUM COMPOUNDS (e.g detergents and soap) result in FALSE DEC/FALSE NEG
SSA interferences: Drugs (Tulbotamide, penicillin, sulfonamide, cephalosporin)
False increase/false pos
*Remember: In the SSA test, only HIGHLY ALKALINE URINE and QUATERNARY AMMONIUM COMPOUNDS (e.g detergents and soap) result in FALSE DEC/FALSE NEG
SSA interferences: Para-amino-salicylic acid/salicylates
false increase/false pos
*Remember: In the SSA test, only HIGHLY ALKALINE URINE and QUATERNARY AMMONIUM COMPOUNDS (e.g detergents and soap) result in FALSE DEC/FALSE NEG
Microscopically, what is the SSA pattern if proteins cause a positive reaction?
Amorphous
Microscopically, what is the SSA pattern if drugs and radiographic contrast dye cause a positive reaction?
Crystalline
T/F: Proteins mainly albumin accepts protons from the indicator
F.
accepts HYDROGEN IONS from the indicator
Which reagent in SSA is acidic?
Multistix (Tetrabromphenol blue) = YELLOW
Which reagent is alkaline in SSA?
Chemstrip (tetrachlorophenoltetrabromosulfonphthalein) = BLUE-GREEN
Sorensen’s test grading: <10
normal
Sorensen’s test grading: 10-29
trace
Sorensen’s test grading: 30-99
1+
Sorensen’s test grading: 100-299
2+
Sorensen’s test grading: 300-1999
3+
Sorensen’s test grading: greater than or equal to 2000
4+
SSA test is a.k.a
EXTON’S TEST
Exton’s test is a.k.a
SSA test
(+) SSA
(-) ALBUMINURIA
A. Albuminuria
B. Proteinuria other than albumin
C. Pak ganern
B.
*Rgnt strip - detects albumin ONLY
SSA - detects all proteins
The most frequently performed chemical analysis on urine (due to monitoring of DM)
GLUCOSE
This is the plasma concentration of a substance at which tubular reabsorption stops
Renal threshold
What is the renal threshold for glucose?
160-180 mg/dL
What specimens are used for glucose parameter in rgnt strip?
-fasting or second morning
-8 hour urine sample or first morning urine
-2 hr post prandial
*Note: A first morning specimen does not always represent a fasting spx because glucose from an evening meal may remain in the bladder overnight, and patients should be advised to empty the bladder and collect the second specimen
2 clinical significance of urine glucose (dalawang dahilan kung bakit may GLUCOSE ang ihi):
1 Hyperglycemia - Associated
2 Renal - associated (impaired tubular REABSORPTION)
Signs of hyperglycemia-associated urine glucose:
-increased blood glucose
-increased urine glucose
Signs of renal-associated urine glucose
-Normal blood glucose
-Increase urine glucose
hyperglycemia-association is caused by:
-DM and Gestational DM
-Pancreatitis and Pancreatic cancer
-Pheochromocytoma
-Acromegaly
- Cushing syndrome
- Hyperthyroidism
- Liver disease
- Cerebrovascular accident / stroke
renal-association is caused by:
-Fanconi’s syndrome
- Advanced renal disease
-Osteomalacia
- Pregnancy
- ESRD (End-stage renal disease)
What is the principle for the rgnt strip reaction for glucose?
double sequential enzymatic reaction
What are the 2 enzymes in the reaction for glucose?
- Glucose oxidase
- peroxidase
Most specific enzyme for B-D-glucose
glucose oxidase
What is the indicator of multistix for glucose in reagent strip?
Glucose oxidase, peroxidase, potassium iodid (blue to green to brown)
what is the indicator substance in a chemstrip?
glucose oxidase, peroxidase, tetramethylbenzidine (yellow to green)
Color produced by Aminopropylcarbazole?
yellow to orange brown
color produced by O-toluidine pink
Pink to purple
Vitamin C can cause FALSE NEGATIVE in what parameters in the rgnt strip?
- Blood
-Bilirubin
-Leukocyte esterase - Nitrite
-Glucose
“BBLNG”
This is added by the manufacturers in the glucose reagent strip to minimize interference by ascorbic acid.
Iodate
This chemical oxidizes ascorbic acid so that it cannot interfere with the oxidation of the chromogen
Iodate
This tablet is used to test for sugars/carbs except sucrose
Clinitest
This tablet is used to test for Bilirubin
Ictotest
This tablet is used to test for ketones
Acetest
Copper reduction test is a.k.a
- BENEDICT’S TEST or CLINITEST
This is a non-specific test for REDUCING sugars such as glucose, galactose, fructose, maltose, lactose, and pentoses.
Copper reduction test
This is a NON-REDUCING sugar and cannot be detected by the Benedict’s test
Sucrose
What is the clin. significance of the Benedict’s test?
For the detection of inborn error of metabolism
This test is clinically significant for the detection of inborn error of metabolism especially galactosuria in newborns in which there is a LACK OF ENZYME galactose-1-phosphate uridyltransferase
Copper reduction test / clinitest / benedict’s test
The copper reduction test is clinically significant for the detection of inborn error of metabolism especially galactosuria in newborns in which there is a LACK OF WHAT ENZYME???
galactose - 1-phosphate uridyltransferase
Component of the tablet: Copper sulfate
main reacting agent
Component of the tablet: sodium carbonate
eliminates interfering O2 (room air)
Component of the tablet: Sodium citrate/citric acid
for heat production
Component of the tablet: sodium hydroxide
for heat production
Component of the tablet: main reacting agent
copper sulfate
Component of the tablet: eliminates interfering O2 (room air)
sodium carbonate
Component of the tablet: for heat production
- sodium citrate/citric acid
- sodium hydroxide
In the copper reduction test, these act as an effervescent
- sodium carbonate
- citric acid
In the Clinitest, this provides the alkaline medium necessary for the reaction, and the heat required is provided by the reaction of sodium hydroxide with water and citric acid
sodium hydroxide
How long is the waiting after the boiling has stopped before gently shaking the contents of the tube in a clinitest?
15 seconds (there is effervescent formation)
T/F: Clinitest tablets are hygroscopic and should be stored in their tightly closed packages.
T
In Clinitest, this color suggests deterioration ue to moisture accumulation, as does vigorous table frizzing
a strong blue color in the unused tablets
this occurs when greater than 2 g/dL sugar is present
Pass through phenomenon
(From a TRUE positive reaction to a FALSE negative reaction due to too much sugar in the urine)
What is the color change that occurs in a pass through phenomenon?
From blue > green > yellow > orange/brick red > green brown
How to prevent pass through phenomenon?
use 2 gtts urine
False neg/false pos interference in Benedict’s test: Contamination of oxidizing agents and detergents
False pos
*In the Benedict’s test there is only ONE interference that results in FALSE POSITIVE (Contamination of oxidizing agents and detergents)
False neg/false pos interference in Benedict’s test: High levels of ascorbic acid
False neg
*In the Benedict’s test there is only ONE interference that results in FALSE POSITIVE (Contamination of oxidizing agents and detergents)
False neg/false pos interference in Benedict’s test: High levels of ketones
false neg
*In the Benedict’s test there is only ONE interference that results in FALSE POSITIVE (Contamination of oxidizing agents and detergents)
False neg/false pos interference in Benedict’s test: High SG
False neg
*In the Benedict’s test there is only ONE interference that results in FALSE POSITIVE (Contamination of oxidizing agents and detergents)
False neg/false pos interference in Benedict’s test: Low temp
False neg
*In the Benedict’s test there is only ONE interference that results in FALSE POSITIVE (Contamination of oxidizing agents and detergents)
False neg/false pos interference in Benedict’s test: Improperlyy preserved specimens
False neg
*In the Benedict’s test there is only ONE interference that results in FALSE POSITIVE (Contamination of oxidizing agents and detergents)
What is the principle in the Benedict’s test?
Copper reduction
The sensitivity of Clinitest to glucose is reduced to how much, therefore, the Clinitest cannot be used as a confirmatory test for glucose
reduced to a minimum of 200 mg/dL
Why can’t the Clinitest be used as a confirmatory test for glucose?
Because the sensitivity of Clinitest to glucose is reduced to a minimum of 200 mg/dL
False pos/false neg interference for Benedict’s test: Vit C
False pos
False pos/false neg interference for Benedict’s test: Formalin
false pos
False pos/false neg interference for Benedict’s test: Uric acid
false pos
False pos/false neg interference for Benedict’s test: cephalosporin
false pos
False pos/false neg interference for Benedict’s test: reducing agents
false pos
False pos/false neg interference for Benedict’s test: oxidizing agents
false neg
False pos/false neg interference for Benedict’s test: detergent
false neg
Benedict’s test grading: Blue
Negative
Benedict’s test grading: Green
1+
Benedict’s test grading: Yellow
2+
Benedict’s test grading: Orange
3+
Benedict’s test grading: Brick red
4+
Sugar level for Benedict’s test: Blue (Negative)
1/4%
*Benedict’s test grading GYOR
Sugar level for Benedict’s: Green (1+)
1/2%
*Benedict’s test grading GYOR
Sugar level for Benedict’s: Yellow (2+)
3/4%
*Benedict’s test grading GYOR
Sugar level for Benedict’s: Orange (3+)
1%
*Benedict’s test grading GYOR
Sugar level for Benedict’s: Brick red (4+)
2%
*Benedict’s test grading GYOR
This is a result from increased fat metabolism.
Ketones
These are formed from beta oxidation of fats
Ketones
Condition associated: Inability to metabolize or utilize available carbohydrate
Type 1 DM
Condition associated: Increased loss of carbohydrates
vomiting
Condition associated: inadequate intake of carbohydrate
starvation and malabsorption/pancreatic disorder
Condition associated: Overuse of available carbohydrates
frequent strenuous exercise
Major ketone but not detected in reagent strip
78% Beta hydroxybutyric acid
PARENT ketone
20% Acetoacetic acid (AAA) / Diacetic acid
Main ketone body detected
20& Acetoacetic acid (AAA) / Diacetic acid
This ketone is detected only when glycine is present
2% Acetone
3 Ketone bodies:
- 78% Beta Hydroxybutyric acid
- 20% Acetoacetic acid / Diacetic acid
- 2% Acetone
2% Acetone is detected only when this substance is present
GLYCINE
What is the renal threshold level for ketones?
70 mg/dL
What happens when the blood ketone concentration exceeds 70 mg/dL?
Ketones are excreted in the urine
For ketones, what color is produced for a positive result?
PURPLE
Reagents in rgnt strip reaction for ketones
- Sodium nitroprusside (nitroferricyanide)
- glycine (chemstrip)
False pos/false neg interferences in ketones: Phthalein dyes
false pos
*in ketones, there is only ONE interference that causes a FALSE NEGATIVE (Improperly preserved specimens)
False pos/false neg interferences in ketones: highly pigmented red urine
false pos
*in ketones, there is only ONE interference that causes a FALSE NEGATIVE (Improperly preserved specimens)
False pos/false neg interferences in ketones: levodopa
false pos
*in ketones, there is only ONE interference that causes a FALSE NEGATIVE (Improperly preserved specimens)
False pos/false neg interferences in ketones: medications containing free sulfhydryl groups
false pos
*in ketones, there is only ONE interference that causes a FALSE NEGATIVE (Improperly preserved specimens)
False pos/false neg interferences in ketones: mercaptoethane sulfonate sodium (MESNA)
false pos
*in ketones, there is only ONE interference that causes a FALSE NEGATIVE (Improperly preserved specimens)
False pos/false neg interferences in ketones: improperly preserved specimens
false neg
*in ketones, there is only ONE interference that causes a FALSE NEGATIVE (Improperly preserved specimens)
The Acetest tablet is composed of:
a-sodium nitroprusside
b-disodium phosphate
c-lactose
this component in the Acetest tablet gives better color differentiation
Lactose
This tablet HAS BEEN used as a confirmatory test for questionable reagent strip results, however, it was primarily used for testing serum and other bodily fluids and dilutions of these fluids for severe ketosis
Acetest
How long must the Acetest test be read?
read for 30 seconds
T/F: Acetest tablets are hygroscopic
T
T/F: Acetst can only be used to test urine.
F.
Acetest can be used to test urine, serum, plasma, or whole blood
T/F: Acetest is 10x more sensitive to diacetic acid than to acetone
T
Any amount of blood greater than how much is considered CLINICALLY SIGNIFICANT
> 5 cells per uL of urine
This shows CLOUDY RED URINE
Hematuria
This shows PRESENCE OF AN INTACT RBC
Hematuria
This produced a SPECKLED/SPOTTED pattern on reagent pad
Hematuria
This shows a CLEAR RED URINE
Hemoglobinuria
This shows a UNIFORM GREEN/BLUE COLOR in reagent strip pad
Hemoglobinuria
This may result from the LYSIS OF RBCs produced in the urinary tract, particularly in dilute, alkaline urine
Hemoglobinuria
This ALSO shows a CLEAR RED URINE
myoglobinuria
This portion of the myoglobin is toxic to the renal tubules
heme portion
This ALSO shows a UNIFORM GREEN / BLUE COLOR in reagent strip pad
Myoglobinuria
Condition associated: Glomerulonephritis
Hematuria
Condition associated: Renal calculi
Hematuria
Condition associated: Pyelonephritis
hematuria
Condition associated: Tumors
hematuria
Condition associated: Trauma
Hematuria
Condition associated: Anticoagulants
Hematuria
Condition associated: Strenuous exercise
hematuria
Condition associated: Hypertension
hematuria
Condition associated: Cystitis
Hematuria
Condition associated: Exposure to toxic chemical
Hematuria
Condition associated: transfusion rxns
hemoglobinuria
Condition associated: Hemolytic anemias
Hemoglobinuria
Condition associated: Severe burns
hemoglobinuria
Condition associated: malaria
hemoglobinuria
Condition associated: syphilis
hemoglobinuria
Condition associated: mycoplasma
hemoglobinuria
Condition associated: C. perfringens
Hemoglobinuria
Condition associated: Strenuous exercise (2)
Hemoglobinuria
Condition associated: Brown recluse spider bites
hemoglobinuria
How much hemoglobin must be present in the urine before it is detected by routine protein reagent strip tests?
EXCEEDING 10 mg/dL
Condition associated: Rhabdomyolysis
Myoglobinuria
Condition associated: Prolonged coma
myoglobinuria
Condition associated: convulsions
myoglobinuria
Condition associated: Extensive exertion
myoglobinuria
Condition associated: muscle wasting diseases
myoglobinuria
Condition associated: cholesterol-lowering statin medications
myoglobinuria
Condition associated: muscle ischemia associated with carbon monoxide poisoning
Myoglobinuria
Condition associated: muscle infection (myositis)
myoglobinuria
Condition associated: trauma (2)
Myoglobinuria
Condition associated: Crush syndrome
Myoglobinuria
Condition associated: ALCOHOLISM
Myoglobinuria
Condition associated: Heroin abuse
myoglobinuria
Reabsorption of filtered hemoglobin also results in the appearance of what?
LARGE YELLOW-BROWN GRANULES OF DENATURED FERRITIN CALLED HEMOSIDERIN
This has an appearance of large yellow-brown granules of DENATURED FERRITIN
Hemosiderin
This is toxic to the renal tubules, and ihgh concentrations can cause acute renal failure
Heme
the heme portion of myoglobin is toxic to the renal tubules, and high concentrations can cause what?
It can cause ACUTE RENAL FAILURE
What are the 2 tests to differentiate hemoglobin and myoglobin?
1-Plasma examination
2-Blondheim’s precipitation test (Ammonium sulfate)
What is the appearance of hemoglobin in PLASMA EXAMINATION?
Red/pink plasma due to hemolysis
What is the appearance of myoglobin in a plasma examination?
Pale yellow plasma
What is the appearance of Hemoglobin in a Blondheim’s precipitation test (Ammonium sulfate)?
Precipitated by ammonium sulfate
What is the appearance of myoglobin in Blondheim’s precipitation test (Ammonium sulfate)?
NOT precipitated by ammonium sulfate
This produces a CLEAR SUPERNATANT that is NEGATIVE for blood rgnt strip
Hemoglobin
This produces a RED SUPERNATANT that is POSITIVE for blood rgnt strip
Myoglobin
What is the color for a NEGATIVE result for blood in rgnt strip?
YELLOW (-)
What is the color for a POSITIVE result for blood in rgnt strip?
(+) Green to Blue
For blood reactions in a rgnt strip, how much concentration of blood can be detected?
Rgnt strip tests can detect concentrations as low as 5 RBCs per uL
Through the acivity of this enzyme of the heme moiety, peroxide is REDUCED and the chromogen becomes OXIDIZED, producing a color change on the reaction pad from YELLOW TO GREEN.
Through pseudoperoxidase
Through pseudoperoxidase activity of the heme moiety, this can be reduced and the chromogen becomes oxidized, producing a color change on the reaction pad from yellow to green
PEROXIDE is reduced
Through pseudoperoxidase activity of the heme moiety, peroxide can be reduced and this becomes oxidized, producing a color change on the reaction pad from yellow to green
CHROMOGEN
Through pseudoperoxidase activity of the heme moiety, peroxidase can be reduced and the chromogen becomes oxidized, producing a color change on the reaction pad of what?
from yellow to green
What is the indicator in MULTISTIX for blood?
diisopropylbenzenedehydroperoxidetetramethylbenzidine
What is the indicator in a CHEMSTRIP for rgnt strip for blood?
dimethyldihydroperoxyhexanetetramethylbenzidine
False pos/false neg interference for blood in rgnt strip: Strong oxidizing agents
False pos
False pos/false neg interference for blood: Vegetable and bacterial peroxidases
false pos
False pos/false neg interference for blood: Escherichia coli
false pos
False pos/false neg interference for blood: menstrual contamination
false pos
False pos/false neg interference for blood: High S.G
False neg
False pos/false neg interference for blood: Crenated cells
false neg
False pos/false neg interference for blood: Formalin
False neg
False pos/false neg interference for blood: Captopril
False neg
False pos/false neg interference for blood: Ascorbic acid ( >25 mg/dL)
False neg
False pos/false neg interference for blood: Unmixed specimen /failure to mix spx prior to testing
False neg
False pos/false neg interference for blood: High concentration of nitrite ( >10mg/dL)
False neg
The appearance of this in the urine can provide an EARLY INDICATION OF LIVER DISEASE
BILIRUBIN (B2)
Hepatic jaundice is associated with what condition/s?
Hepatitis and Cirrhosis
Post-Hepatic jaundice is associated with what condition/s?
Biliary obstruction (gallstones, carcinoma)
This is associated with Heptitis and cirrhosis
Hepatic jaundice
this is associated with biliary obstruction
Post hepatic jaundice
What bilirubin type is WATER SOLUBLE thus can be seen in urine and can be detected
ONLY B2 or CONJUGATED bilirubin
T/F: B2 is insoluble in water
F.
Water soluble
It produces an amber urine with yellow foam
B2
What color urine does B2 produce?
AMBER URINE
What color foam does B2 produce?
YELLOW foam
*mukhang yellow yung tao pag mataas ang bilirubin or may hepatitis
Conjugated bilirubin is normally excreted where?
excreted in the bile
Conjugated bilirubin is normally excreted in the bile into the what?
into the duodenum
Normal adult urine contains only how much of bilirubin per deciliter?
0.02 mg of bilirubin per deciliter
Excretion of bilirubin is enhanced by what?
enhanced by alkalosis
This is a HIGHLY pigmented YELLOW compoun
Bilirubin
this is a degradation product of hemoglobin
Bilirubin
What is the NORMAL lifespan of RBCs?
approx. 120 days
What organ/s destroys RBCs after 120 days?
spleen and liver by phagocytic cells of the reticuloendothelial system
Free hemoglobin is broken down into what components?
- iron
- protein
- protoporphyrin
T/F: The body reuses the iron and protein, and the cells of the reticuloendothelial system convert the remaining protoporphyrin to bilirubin
T
What is the indicator in the rgnts used for rgnt strip reaction for Bilirubin? (Multistix)
2,4-dichloroaniline diazonium salt
What is the indicator in the rgnts used for rgnt strip reaction for Bilirubin? (Chemstrip)
2,6-dichlorobenze diazonium tetrafluoroborate
False pos/false neg interference for Bilirubin: Highly pigmented urines
False pos
False pos/false neg interference for Bilirubin: phenazopyridine
false pos
False pos/false neg interference for Bilirubin: indican
false pos
False pos/false neg interference for Bilirubin: metabolites of Iodine
false pos
False pos/false neg interference for Bilirubin: specimen exposure to light
false neg
False pos/false neg interference for Bilirubin: ascorbic acid
false neg
False pos/false neg interference for Bilirubin: high concentration of nitrite
false neg
This parameter in the rgnt strip is more difficult to interpret than other rgnt strip rxns and are EASILY INFLUENCED by other pigments present in the urine
Rgnt strip color rxns for BILIRUBIN
This is the confirmatory test for bilirubin
Ictotest
Ictotest is able to detect how much of bilirubin
as little as 0.05 mg/dL
This test is much more sensitive than the dipsticks
ictotest
Components of Ictotest:
1 p-nitrobenzene-diazonium p-toluenesulfonate
2 SSA
3 Sodium carbonate
4 Boric acid
Positive reaction in Ictotest:
BLUE to PURPLE color
This is a bile pigment that results from hemoglobin degradation
UROBILINOGEN
Conjugated bilirubin is reduced by intestinal bacteria into what?
urobilinogen
This is REDUCED by intestinal bacteria into urobilinogen
conjugated bilirubin
How much of urobilinogen is NORMALLY found in the urine
less than 1 mg/dL or Ehrlich unit
T/F: Constipation can raise urobilinogen level
T
*1% of the non-hospitalized population and 9% of a hospitalized populatopm exhibit elevated results.
What is the PRINCIPLE for Urobilinogen in the rgnt strip?
EHRLICH’S REACTION
In urobilinogen, what indicator is used in Multistix?
Ehrlich reagent
in urobilinogen, what indicator is used in chemstrip?
4-methyloxybenzene-diazonium-tetrafluoroborate (more SPECIFIC than ehrlich’s rxn)
What are the ehrlich-reactive compoounds?
- porphobilinogen
- indican
- p-aminosalicylic acid
- sulfonamides
- methyldopa
- chlorpromazine
*Also gives POSITIVE RXN for Ehrlich’s rxn
false pos/false neg interference in Urobilinogen: other ehrlich’s compound
False pos
*In urobilinogen, there are only TWO interferences that cause a false positive: other ehrlich’s compound and highly pigmented urine
false pos/false neg interference in Urobilinogen: highly pigmented urine
false pos
*In urobilinogen, there are only TWO interferences that cause a false positive: other ehrlich’s compound and highly pigmented urine
false pos/false neg interference in Urobilinogen: old spx
false neg
*In urobilinogen, there are only TWO interferences that cause a false positive: other ehrlich’s compound and highly pigmented urine
false pos/false neg interference in Urobilinogen: preservation in formalin
false neg
*In urobilinogen, there are only TWO interferences that cause a false positive: other ehrlich’s compound and highly pigmented urine
false pos/false neg interference in Urobilinogen: improperly preserved, allowing urobilinogen to be photo-oxidized to urobilin
false neg
*In urobilinogen, there are only TWO interferences that cause a false positive: other ehrlich’s compound and highly pigmented urine
false pos/false neg interference in Urobilinogen: high concentration of nitrite
false neg
*In urobilinogen, there are only TWO interferences that cause a false positive: other ehrlich’s compound and highly pigmented urine
What would be the result of urobilinogen measurements if the rgnt strip is performed at a higher temperature?
FALSELY INCREASED
*The sensitivity of the Ehrlich rxn INCREASES with temperature, and testing should be performed at RT
What would be the result of urobilinogen measurements after following a meal?
normally highest after meal
*as a result of increased excretion of bile salts, urobilinogen results are normally highest following a meal
Used to differentiate urobiliinogen, porphobilinogen, and other Ehrlich reactive compounds
Watson-Schwartz test
This test uses extraction with organic solvents chloroform and butanol
Watson-Schwartz test
Watson-Schwartz test uses extraction with organic solvents like?
chloroform and butanol
Using chloroform extract, what is usually settled at the bottom at remains at the top, respectively?
- Chloroform (bottom)
- Urine (top)
*UC BU
Using butanol extract, what is usually settled at the bottom at remains at the top, respectively?
- urine (bottom)
- butanol (top)
*UC BU
Using chloroform extract, what is usually settled at the bottom at remains at the top, respectively in UROBILINOGEN?
- Red (chloroform;bottom)
- Colorless (urine;top)
Using chloroform extract, what is usually settled at the bottom at remains at the top, respectively in PORPHOBILINOGEN?
- Colorless (chloroform; bottom)
- Red (urine;top)
Using chloroform extract, what is usually settled at the bottom at remains at the top, respectively in OTHER EHRLICH REACTIVE COMPOUNDS?
- Colorless (chloroform; bottom)
- Red (Urine; top)
Using butanol extract, what is usually settled at the bottom at remains at the top, respectively in PORPHOBILINOGEN?
- Red (urine; bottom)
- Colorless (Butanol;top)
Using butanol extract, what is usually settled at the bottom at remains at the top, respectively in UROBILINOGEN?
- Urine (colorless; bottom)
- Butanol (red;top)
Using butanol extract, what is usually settled at the bottom at remains at the top, respectively in OTHER EHRLICH REACTIVE COMPOUND?
- Colorless (urine;bottom)
- Red (Urine; top)
This is a.k.a as INVERSE EHRLICH REACTION
Hoesch test
This is a rapid screening test for porphobilinogen (>2 mg/dL)
Hoesch test (inverse ehrlich rxn)
What is the appearance of a positive Hoesch test?
RED on TOP of solution
In the Hoesch test, how much ehrlich’s reagent is dissolved?
6M or 6N HCl
The Hoesch test can detect how much porphobilinogen?
approx. 2 mg/dL
Urobilinogen is inhibited by what?
HIGHLY acidic pH
T/F: Urobilinogen is inhibited by highly alkaline pH
F.
Highly acidic pH
In the Hoesch test, these may produce FALSE POSITIVE results
High concentrations of methyldopa and indican, and highly pigmented urine
Hemoytic disease is associated with kind of jaundice?
Prehepatic jaundice
Liver damage is associated with what kind of jaundice?
Hepatic jaundice
Bile duct obstruction is associated with what kind of jaundice?
post hepatic jaundice
What is the result of BLOOD in prehepatic jaundice?
Increase unconjugated bilirubin
what is the result of BLOOD in hepatic jaundice?
Increase both B1 and B2
What is the result of blood in post hepatic jaundice?
increase conjugated bilirubin
What disease is assoc. with prehepatic jaundice
hemolytic disease
what disease is assoc with hepatic jaundice?
liver damage
what disease is associated with post hepatic jaundice?
bile duct obstruction
what is the result of urine bilirubin in pre hepatic jaundice?
NEGATIVE
What is the result of urine bilirubin in hepatic jaundice?
+/-
what is the result of urine bilirubin in post hepatic jaundice??
+++
what is the result of urine urobilinogen in prehepatic jaundice?
+++
what is the result of urine urobilinogen in hepatic jaundice?
++
what is the result of urine urobilinogen in post hepatic jaundice?
Normal
This provides a rapid screening test for the presence of UTI and bacteriuria
Nitrite
this can also be used to evaluate the sucess of antibiotic therapy
nitrite test
this test can be used to periodically screen persons with recurrent infections, patients with diabetes, and pregnant women, all of whom are considred to be at high risk for UTI
Nitrite test
T/F: Nitrite test is intended to replace the urine culture as the primary test for diagnosing and monitoring bacterial infection
F.
NOT INTENDED TO REPLACE
What is the specimen used for nitrite test?
1st morning or 4-hour urine
What is the chemical basis of the nitrite test?
ability of certain bacteria to reduce nitrAte (a normal constituent of urine) to nitrIte (which does not normally appear in the urine
Nitrite at an ACIDIC pH reacts with an aromatic amine called ???
para-arsanilic acid or sulfanilamide
Nitrite at an acific pH reacts with an aromatic amine to form a DIAZONIUM compound that then reacts with TETRAHYDROBENZOQUINOLIN compounds to produce what COLOR??
Pink-colored azodye
Nitrite reacts with sulfanilamide to form a diazonium compound that then reacts with what compound to produce a pink-colored azodye?
TETRAhydroBENZOquinolin
For nitrite determination in a rgnt strip, Multistix uses what indicator?
p-arsanilic acid, tetrahydrobenzoquinolin-3-ol
*Si GREISS (Grace) mahilig sa pink
For nitrite determination in rgnt strip, Chemstrip uses these indicators:
Sulfanilamide, HYDROXYtetraHYDRObenzoQUINOLINE
False pos/false neg for nitrite determination in rgnt strip: Improperly preserved spx
false pos
False pos/false neg for nitrite determination in rgnt strip: highly pigmented urine
false pos
False pos/false neg for nitrite determination in rgnt strip: non reductase containing bacteria
false neg
False pos/false neg for nitrite determination in rgnt strip: insufficient contact time between bacteria and urinary nitrate
false neg
False pos/false neg for nitrite determination in rgnt strip: large quantities of bacteria converting nitrite to nitrogen
false neg
False pos/false neg for nitrite determination in rgnt strip: presence of antibiotics
false neg
False pos/false neg for nitrite determination in rgnt strip: ascorbic acid
false neg
False pos/false neg for nitrite determination in rgnt strip: high S.G
false neg
*For Nitrite, there are only 2 interferences that show a FALSE POSITIVE result (Improperly preserved spx and highly pigmented urine)
Positive result for nitrite should appear as:
UNIFROM/HOMOGENOUS PINK
How is nitrite in rgnt strip reported as?
reported only as negative or positive
How will you report the appearance of PINK SPOTS/EDGE in a nitrite rgnt strip?
NEGATIVE
This test is for gram negative bacteria/bacilli which are mostly nitrite positive
Hoesch test
This bacteria gives nitrite positive results
Enterobacteriaceae/coliform
This rgnt strip parameter is signficant in UTI/Inflammation, screening of urine culture specimen, bacterial and non-bacterial infection
Leukocytes
It detects the presence of leukocytes that have been lysed, particularly in dilute ALKALINE urine
Leukocytes
It offers a more standardized means for detection of leukocytes
Leukocytes in a rgnt strip
T/F: In a reagent strip test for leukocytes, it is not design to measure the concentration of leukocytes
T
T/F: For leukocytes, it is recommended that quatitation should be done by microscopic examination
T
In an LE test, this is detected in Neutrophil, Basophil, Eosinophil, Monocytes, Trichomonas, Chlamydia, Yeast, and Histocytes
esterase
An LE test detects esterase found in these cells:
-neutrophil
- basophil
-eosinophil
- monocytes
-trichomonas
-chlamydia
-Yeast
-Histocytes
T/F: An LE test is NEGATIVE for lymphocytes
T
Screening urine specimens using LE test should be correlated with what?
correlated with nitrite chemical rxns
These cells do NOT contain esterases:
- Lymphocytes
- Erythrocytes
-Bacteria - renal tissue cells
These infections cause leukocytura or pyuria without bacteriuria
-trichomonads
-mycoses (e.g., yeast)
-chlamydia
-mycoplasmas
- viruses
- tuberculosis
What is the principle for rgnt strip reaction for leukocytes?
LEUKOCYTE ESTERASE
This is the appearance of a POSITIVE reaction in rgnt strip for leukocytes
Purple azodye
For leukocytes, Multistix uses these indicators?
Diazonium salt, derivatized pyrrole amino acid ester
For leukocyte, Chemstrip uses these indicators:
- Diazonium salt, indoxylcarbonic acid ester
Sensitivity of Multistix for Leukocytes:
5 to 15 WBC/hpf
Sensitivity of Chemstrip to Leukocytes
10 to 25 WBC/hpf
False pos/false neg interferences for Leukocytes: Strong oxidizing agents
False pos
False pos/false neg interferences for Leukocytes: formalin
false pos
False pos/false neg interferences for Leukocytes: Highly pigmented urine, nitrofurantoin, beets, phenazopyridine
false pos
False pos/false neg interferences for Leukocytes: spx contaminated with vaginal secretions
false pos
False pos/false neg interferences for Leukocytes: high concentration of protein (>500 mg/dL)
false neg
False pos/false neg interferences for Leukocytes: High glucose (greater than or equal to 3 g/dL)
false neg
False pos/false neg interferences for Leukocytes: oxalic acid (in acidified urine that has 4.4 pH or below)
False neg
False pos/false neg interferences for Leukocytes: ascorbic acid
false neg
False pos/false neg interferences for Leukocytes: gentamicin
false neg
False pos/false neg interferences for Leukocytes: cephalosporins
false neg
False pos/false neg interferences for Leukocytes: tetracyclines
false neg
False pos/false neg interferences for Leukocytes: inaccurate timing
false neg
This test requires the loooooongest time of all the reagent strip reactions (2 mins)
LE rxn
This is the 11th parameter in a reagent strip test
Ascorbic acid
Ascorbic acid causes a FALSE NEGATIVE reaction to these parameters:
-Blood
-Bilirubin
- Leukocyte
-Nitrite
- Glucose
*BBLNG
Ascorbic acid causes a false positive result to this tablet
Clinitest
This ascorbic level causes a negative reaction to Bilirubin and nitrite
greater than or equa to 25 mg/dL
This ascorbic acid level causes a negative reaction to glucose
greater than or equal to 50 mg/dL
An ascorbic acid level of greater than or equal to 50 mg/dL causes what reaction to glucose
causes a negative reaction
An ascorbic acid level of greater than or equal to 50 mg/dL causes a negative reaction to what?
negative reaction to glucose
An ascorbic acid level of greater than or equal to 25 mg/dL causes what reaction to bilirubin and nitrite
negative reaction
An ascorbic acid level of greater than or equal to 25 mg/dL causes a negative reaction to what?
Bilirubin and nitrite
What brand for Ascorbic acid has a reading time of 60 seconds?
Stix
What brand for Ascorbic acid has a reading time of 10 seconds?
C-six
What brand for Ascorbic acid produces a blue color for positive?
BOTH stix and c-stix
What are the 2 brands for Ascorbic acid?
- Stix
- C-stix
The brand Stix for ascorbic acid has a reading time of what?
60 seconds
The brand C-Stix for ascorbic acid has a reading time of what?
10 seconds
What is the principle of Bilirubin test in rgnt strip?
Diazo rxn
What is the principle of Glucose test in rgnt strip?
Double sequentioal enzymatic rxn
What is the principle of Ketones test in rgnt strip?
Sod. Nitroprusside (Legal’s reaction)
What is the principle of S.G test in rgnt strip?
pKa change of polyelectrolyte
What is the principle of pH test in rgnt strip?
Double indicator system
What is the principle of Protein test in rgnt strip?
Protein (Sorensen’s) error of indicator
What is the principle of Blood test in rgnt strip?
Pseudoperoxidase activity of heme
What is the principle of Urobilinogen test in rgnt strip?
Ehrlich’s rxn
What is the principle of Nitrite test in rgnt strip?
Greiss rxn
What is the principle of Leukocyte test in rgnt strip?
Leukocyte esterase
This uses the principle: Diazo rxn
bilirubin
This uses the principle: Double sequential enzymatic rxn
Glucose
This uses the principle: Sod. Nitroprusside (Legal’s rxn)
Ketones
This uses the principle: pKa change of polyelectrolyte
S.G
This uses the principle: Double indicator system
pH
This uses the principle: Protein (Sorensen’s) error of indicator
Protein
This uses the principle: Pseudoperoxidase activity of heme
Blood
This uses the principle: Ehrlich’s reaction
Urobilinogen
This uses the principle: Greiss rxn
Nitrite
This uses the principle: Leukocyte esterase
Leukocyte
What is the appearance of a positive result in Bilirubin?
Violet, tan, or pink
What is the appearance of a positive result in Glucose
Potassium iodide = blue-green to brown
What is the appearance of a positive result in Ketones
Purple
What is the appearance of a positive result in S.G?
Diluted = blue
concentrated = yellow
What is the appearance of a positive result in pH
Acidic = red to yellow
Alkaline = green to blue
What is the appearance of a positive result in protein
Blue-green
What is the appearance of a positive result in Blood
Green to blue
What is the appearance of a positive result in urobilinogen
Red
What is the appearance of a positive result in Nitrite
pink
What is the appearance of a positive result in leukocyte
purple
In this parameter, it appears as violet, tan, or pink when positive
Bilirubin
In this parameter, it appears as blue-green to brown when positive
glucose
In this parameter, it appears as purple when positive
Ketones and Leukocyte
In this parameter, it appears as blue when diluted and yellow when concentrated when positive
S.G
In this parameter, it appears as red to yellow when acidic and green to blue when alkaline when positive
pH
In this parameter, it appears as blue-green when positive
protein
In this parameter, it appears as green to blue when positive
blood
In this parameter, it appears as red when positive
urobilinogen
In this parameter, it appears as pink when positive
nitrite
What is the reading time for bilirubin?
30 secs
What is the reading time for glucose?
30 s
What is the reading time for ketones?
40 s
What is the reading time for S.G?
45 s
What is the reading time for pH?
60 s
*PPBUN (tests with 60 s as reading time)
pH, Blood, Protein, Urobilinogen, Nitrite
What is the reading time for protein?
60 s
*PPBUN (tests with 60 s as reading time)
pH, Blood, Protein, Urobilinogen, Nitrite
What is the reading time for blood?
60 s
*PPBUN (tests with 60 s as reading time)
pH, Blood, Protein, Urobilinogen, Nitrite
What is the reading time for urobilinogen?
60 s
*PPBUN (tests with 60 s as reading time)
pH, Blood, Protein, Urobilinogen, Nitrite
What is the reading time for nitrite?
60 s
*PPBUN (tests with 60 s as reading time)
pH, Blood, Protein, Urobilinogen, Nitrite
What is the reading time for leukocyte?
120 s
Reducing agent such as vit. C result in false pos for:
- false pos Clinitest
Reducing agent such as vit. C result in false neg in:
- BBLNG
(Blood, Bilirubin, Leukocyte, Nitrite, Glucose)
Oxidizing agents such as detergent or soap result in a false pos result in:
LGBP
-LE
-Glucose
- Blood
-protein
Oxidizing agents such as detergent or soap result in a false negative result in:
- Clinitest
Formalin result in a false pos result in:
- LE
Formalin result in a false neg result in:
Blood and Urobilinogen
Highly pigmented urine result in a false pos result in:
BULKN
(Bilirubin, Urobilinogen, LE, Ketones, Nitrite)
Highly pigmented urine result in a false neg result in:
NONE
Which of the following is NOT a component of clinitest?
a- sodium carbonate
b- lactose
c- copper sulfate
d-Sodium citrate/citric acid
B. Lactose
T/F: Vit. C causes a false positive in Clinitest
True
How many drops of urine must be added in the case of a pass through phenomenon?
2 gtts
T/F: Whenever albumin is present in the urine, the medium in protein will become alkaline (yellow to blue)
T
What is the buffer used for protein rgnt strip?
citric acid / citrate (pH of 3)
The most predominant type of ketone
78% Beta hydroxybutyric acid
Major ketone detected in the urine
20% AAA
What parameters give a PURPLE POSITIVE RXN?
-Bilirubin
- Ketones
- LE
*ang BKL (bakla) mahilig sa purple
This is the confirmatory test in a rgnt strip for Ketones
Acetest
This test is the LEAST sensitive to time
LE
How to differentiate hemoglobinuria vs myoglobinuria?
- Plasma examination
- Blondheim’s precipitation test
how much ammonium sulfate is added to a Blondheim’s precipitation test?
2.8 g
In the pseudoperoxidas activity of heme, which of the following is oxidized?
a- hydrogen peroxide
b-chromogen
c-H2O
B. Chromogen
What color is the reduced form of the chromogen in pseudoperoxidase reaction?
YELLOW
What color is the oxidized form of the chromogen in pseudoperoxidase reaction?
Green to Blue
What form of bilirubin is seen in the urine?
B2
The most difficult parameter to interpret
Bilirubin
most affected parameter in unmixed urine sample
Blood
most affected parameter using a refrigerated sample
Glucose
What is the color of urobilinogen?
Colorless
Urobilin is a.k.a
Stercobilin (for stool)
When conjugated bilirubin (B2) is reduced it forms what?
Urobilinogen
When urobilinogen is OXIDIZED, it forms what?
Urobilin or stercobilin (pigment that gives urine or stool its color)
What is the ratio of urobilinogen to Ehrlich’s to mg/dL?
1:1
*6 mg/dL of urobilinogen is equivalent to how many Ehrlich’s unit?
Ans: 6 Ehrlich’s unit
In the Watson-Schwartz test, which is soluble to both chloroform and butanol?
Urobilinogen
In Watson-Schwartz test, which is insoluble to both chloroform and butanol?
Porphobilinogen
In Watson-Schwartz test, which is soluble to butanol ONLY?
Other Ehrlich reactive compounds
This is a supplemental test for UTI and bacteriuria
Nitrite
Positive nitrite corresponds to _____ organisms/mL
100,000 organisms/mL (minimum amount of microorganism in order to produce a nitrite reaction)
All enterobacteriaceae are nitrite positive except for ???
Shigella and Klebsiella and some Proteus spp
Diazonium salt used as reagent
-bilirubin
-urobilinogen
-LE
*BUL
Diazonium salt as product formed from a reaction
NITRITE REAGENT STRIP
What enzyme converts nitrate to nitrite?
Nitrate reductase
What WBC that does not have esterase activity?
LYMPHOCYTES
T/F: Leukocyte is lysed in dilute acidic urine
F.
Dilute alkaline urine
TIPS FOR REAGENT STRIP REAGENTS
Bilirubin: Dichloro…….. diazonium salt
Blood: Di…….. tetramethylbenzidine
Protein: Tetra…….
S.G: …… bromthymol blue
Nitrite: …… quinolin
