MIDTERM LEC: CHEMICAL EXAMINATION Flashcards
- composed of several chemical-impregnated
- absorbent pads attached to a plastic
- color producing chemical reaction takes place when absorbent pad comes in contact with urine
- reactions are interpreted by comparing the color produced on the pad with a chart supplied by the manufacturer
REAGENT STRIP
TYPES OF REAGENT STRIP
- MULTISTIX
- CHEMSTRIP
(Siemens Healthcare Diagnostics, Deerfield, IN)
MULTISTIX
(Roche Diagnostics, Indianapolis, IN)
CHEMSTRIP
PROCEDURE:
- Dip the reagent strip completely in the urine
- Remove excess urine from the strip by running the edge of the strip on the container when withdrawing it from the specimen
- Blot the strip horizontally on an absorbent medium pad
- Wait for the specified length of time for reactions to take place
- Compare the colored reactions against the
manufacturer’s chart using a good light source
ERRORS CAUSED BY IMPROPER TECHNIQUE
✓ Formed elements such as RBC & WBC sink to the bottom of the specimen therefore it will be undetected in an unmixed specimen
✓ Allowing the strip to remain in the urine for an extended period may cause leaching of reagents from the pads
✓ Excess urine remaining on the strip after its removal from the specimen can produce over-run between chemicals on adjacent pads, producing distortion of the colors
✓ The timing of reactions should be followed; when precise timing can’t be achieved, the reactions
✓ should be read between 60 & 120 second
✓ A good light source is essential for accurate
interpretation of color reactions
✓ The strip must be held close to the color chart without actually being placed on the chart
✓ Reagent strips & color charts from different
manufacturers are not interchangeable
✓ Specimens that have been refrigerated must allow to return to room temperature prior to reagent strip testing
HANDLING & STORAGE OF REAGENT STRIP
✓ Reagent strips must be protected from deterioration caused by moisture, heat, light, & volatile chemicals
✓ Reagents are packed in opaque containers with a desiccant to protect them from light & moisture
✓ Strips are removed just prior to testing, & the bottle is tightly resealed immediately
✓ Bottle should not be open in the presence of volatile fumes
✓ Reagents should be stored at room temperature but never refrigerated
✓ All bottles are stamped with an expiration date that represents the functional life expectancy of the reagent strips therefore it must not be used past the expiration date
✓ Care must be taken not to touch the chemical pads when removing the strips
✓ A visual inspection should be done each time a strip is used to detect deterioration even though the strips may still be within the expiration date
QUALITY CONTROL OF REAGENTS STRIPS
✓ Reagent strips must be checked with both positive & negative controls a minimum of once every 24hrs, usually every start of the shift
✓ Testing is also performed when a new bottle is opened, questionable results are obtained, or when there is concern about the integrity of the reagent strip
AUTOMATED REAGENT STRIP READERS
PRINCIPLE: REFLECTANCE PHOTOMETRY
light reflection from the test pads decreases in
proportion to the intensity of color produced by the concentration of the test substance
PRINCIPLE: REFLECTANCE PHOTOMETRY
The most common sugar in the urine
GLUCOSE
should not be detected in normal urine
GLUCOSE
Small amounts when present are detected by any sensitive test
GLUCOSE
GLUCOSE RENAL THRESHOLD:
160-180 mg/dl
commonly used as a screening test for possible DM or to monitor control of blood glucose in diabetes
GLUCOSE
Other Sugars in Urine
* Identified by TLC:
- FRUCTOSE (Levulose)
- GALACTOSE
- LACTOSE (Glu+Gal)
- PENTOSE
- SUCROSE (NON-REDUCING SUGAR)
increase fruits, honey, syrup,
fructose intolerance
FRUCTOSE(Levulose)
increase in infants with galactosemia
GALACTOSE
increase during pregnancy, lactation, strict milk diet, lactose intolerance
LACTOSE (Glu+Gal)
– increase in benign essential pentosuria (Xylulose, Arabinose)
PENTOSE
increase in intestinal disorders, sucrose intolerance
SUCROSE (non-reducing sugar)
HYPERGLYCEMIA-ASSOCIATED
blood glucose:
urine glucose:
INCREASE; INCREASE
RENAL-ASSOCIATED
blood glucose:
urine glucose:
NORMAL; INCREASE
HYPERGLYCEMIA-ASSOCIATED CAUSES:
- DM
- PANCREATITIS
- ACROMEGALY
- CUSHING’S SYNDROME
- HYPERTHYROIDISM
- PHEOCHROMOCYTOMA
- STRESS
- GESTATIONAL DIABETES
RENAL-ASSOCIATED CAUSES:
- FANCONI SYNDROME
- ADVANCED RENAL DISEASE
- OSTEOMALACIA
- PREGNANCY
GLUCOSE REACTION TIME:
30 SECONDS
GLUCOSE PRINCIPLE:
DOUBLE SEQUENTIAL ENZYME
REACTION
REAGENTS FOR GLUCOSE
MULTISTIX:
Glucose oxidase and Peroxidase
REAGENTS FOR GLUCOSE
MULTISTIX
CHROMOGEN:
Potassium Iodide →
(𝑏𝑙𝑢𝑒 𝑡𝑜 𝑔𝑟𝑒𝑒𝑛 𝑡𝑜 𝑏𝑟𝑜𝑤𝑛)
REAGENTS FOR GLUCOSE
CHEMSTRIP:
Glucose oxidase and Peroxidase
REAGENTS FOR GLUCOSE
CHEMSTRIP
CHROMOGEN:
Tetramethylbenzidine →
(𝑦𝑒𝑙𝑙𝑜𝑤 𝑡𝑜 𝑔𝑟𝑒𝑒𝑛)
REAGENTSTRIP FOR GLUCOSE
FALSE-POSITIVE:
Contamination by oxidizing
REAGENTSTRIP FOR GLUCOSE
FALSE – NEGATIVE:
✓ high levels of ascorbic acid
✓ high levels of ketones
✓ high specific gravity
✓ low temperatures
✓ improperly preserved specimens
COPPERREDUCTION TEST (CLINITEST (TABLET)/BENEDICT’S TEST) PRINCIPLE:
Copper Reduction
ability of the glucose and other substances to
reduce copper sulfate to cuprous oxide in the presence of alkali and heat.
COPPERREDUCTION TEST (CLINITEST (TABLET)/BENEDICT’S TEST)
color change progressing from negative blue
through green, yellow, and orange/red occurs when reaction takes place
COPPERREDUCTION TEST (CLINITEST (TABLET)/BENEDICT’S TEST)
- NOTE: Observe the reaction closely , because at high glucose level “ pass through” phenomenon may occur.
COPPERREDUCTION TEST (CLINITEST (TABLET)/BENEDICT’S TEST)
COPPERREDUCTION TEST (CLINITEST (TABLET)/BENEDICT’S TEST)
PREVENT PASS THROUGH :
use 2 gtts urine
FALSE-POSITIVE : Reducing agents
- Clinitest tablet test follow the same principle with Benedict’s Test
- The classic Benedict’s solution was developed in 1908 and contained copper sulfate, sodium carbonate, and sodium citrate buffer
GLUCOCE OXIDASE: 1+ POSITIVE
CLINETEST: ______
INTERPRETATION: ________
CLINETEST: negative
INTERPRETATION: small amount of glucose
GLUCOCE OXIDASE: NEGATIVE
CLINETEST: ______
INTERPRETATION: __
CLINETEST: positive
INTERPRETATION: non-glucose reducing substance possible interfering substances for reagent strip
GLUCOCE OXIDASE: 4+ POSITIVE
CLINETEST: ______
INTERPRETATION: __
CLINETEST: negative
INTERPRETATION: oxidizing agent interfere on reagent strip
DETECT: Conjugated Bilirubin(water soluble)
BILIRUBIN
provide an early indication of liver disease
BILIRUBIN
pigmented yellow compound from the breakdown of hemoglobin
BILIRUBIN
Amber urine (Tea Color)with yellow foam
BILIRUBIN
CLINICAL SIGNIFICANCE OF BILIRUBIN IN URINE:
- Hepatitis
- Cirrhosis
- Other liver disease
- Biliary obstruction(gallstones, carcinoma)
REAGENTSTRIPFORBILIRUBIN(30 SECONDS)
PRINCIPLE:
Based on the “Diazo Reaction” of Bilirubin
BILIRUBIN
MULTISTIX:
2,4- dichloroaniline diazonium salt
BILIRUBIN
CHEMSTRIP:
2,6- dichlorobenzene diazonium
BILIRUBIN
FALSE- POSITIVE:
✓ Highly pigmented urines
✓ Phenazopyridine
✓ Indican
✓ Metabolites of iodine
BILIRUBIN
FALSE-NEGATIVE:
✓ Specimen exposure to light
✓ High conc. of nitrite
✓ Ascorbic acid
- More sensitive than reagent strip with less interference
- (+) blue to purple color
Contains:
✓ P- nitrobenzene-diazonium Ptoluenesulfonate
✓ SSA
✓ Sodium Carbonate
✓ Boric Acid
ICTOTEST (TABLET)
Result from increased fat metabolism due to inability to metabolize carbohydrates
KETONES
end product of rapid or excessive fatty acid breakdown when glucose is not used as source of energy
KETONE BODIES
KETONE BODIES:
- Beta-hydroxybutyric acid(78%)
- Acetoacetic acid(AAA)/Diacetic
acid (20%)
Acetone(2%)
major ketone but not detected in
reagent strip
Beta-hydroxybutyric acid(78%)
parent ketone
Acetoacetic acid(AAA)/Diacetic
acid (20%)
detected only when glycine is present
Acetone(2%)
CLINICAL SIGNIFICANCE OF KETONES IN URINE:
- Type 1 DM
- Insulin Dosage Monitoring
- Starvation
- Strenuous Exercise
- Malabsorption
- Vomiting
REAGENTSTRIP FOR KETONES(40 SECONDS)
PRINCIPLE:
Based on “NITROPRUSSIDE” reaction for ketones
KETONES REAGENT
CHEMSTRIP:
Sodium Nitroprusside(Sodium
Nitroferricyanide), Glycine
The ketone pad on the multi-reagent dip stick
detects mainly acetoacetic acid and acetone;
_________ is not detected.
ß-hydroxybutyrate
KETONES
FALSE-POSITIVE:
✓ Pthalein dyes
✓ Highly pigmented red urine
✓ Levodopa
✓ Medications containing free sulfhydryl groups
KETONES
FALSE- NEGATIVE:
✓ Improperly preserved specimens
CONTAINS:
✓ Sodium Nitroprusside
✓ Disodium Phosphate
✓ Lactose
ACETEST (TABLET)