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Composition of Reagent Strips
Chemical impregnated absorbent pads attached to plastic strip Color producing chemical reaction takes place when urine comes in contact with strip Reactions are compared with manufacture·s color chart or read by automated instrumentation. Qualitative and semi quantitative results http://www.vet.uga.edu/vpp/CLERK/Sine/
Handling, Storage, and QC of strips
Protect from moisture, heat, volatile chemicals, light and fingerprints Date bottle when opened. Do not use past expiration date or 6 months after opening; Store in original container at room temp. Check strips with positive and negative manufactured controls at least once every 24 hours or each shift.
Dipstick Parameters Clinical Significance Chemical reactions Disease states .
pH Used more to verify crystal identification Normal first morning urine is slightly acidic Helps to rule out suspected acid/base disturbance Used to identify improperly preserved urine .
pH Chemicals and Reagent Strip Reaction Double indicator system ² Methyl red and Bromthymol Blue Methyl Red active from pH 4-6 (orange>>>yellow) Bromthymol Blue active from pH of 6-9 (green>>>blue) .
.Protein Most indicative of renal disease above all other parameters Normally very little protein in urinelow molecular weight proteins that filter through glomerulus Proteinuria can be pre-renal. or post renal in origin. renal.
.Prerenal Proteinuria Not indicative of renal disease Increase of protein prior to reaching kidney Renal threshold is reached and flows into urine Usually low molecular weight proteins not detected by strip (strip detects albumin) Infection and inflammation (hemoglobin. Confirm this with serum protein electrophoresis (monoclonal gammopathy). myoglobin. acute phase reactants) Multiple Myeloma ²Malignant proliferation of B cells. Bence Jones Protein ² coagulates in urine between 40-600 C and clears at 1000 C.
immune complexes ² i.strenuous exercise. toxic substances. dehydration. microalbumin (routine strip does not pick up microalbumin) is good parameter to measure to predict diabetic complications. . lupus or streptococcal AGN Increased blood pressure entering glomerulus. II ² complication of diabetes. tubular dysfunction Diabetes mellitus ² Type I. hypertension.Renal Proteinuria Glomerular or tubular damage.from amyloid material.e. pregnancy (preeclampsia).
Postrenal Proteinuria Protein added to urine as passes through lower urinary track (ureters. vagina) Bacterial or fungal infections Bleeding as a result of injury or menstrual contamination . bladder. urethra.
exposure to cold. Transient ² strenuous exercise. then positive protein in urine after standing a few hours. Seen in young adults. dehydration . high fever.Benign Proteinuria Persistent ² Orthostatic Proteinuria ² increased pressure on renal vein when in upright position (vertical). negative first morning protein.
radiographic dye. false pos. false neg ² highly alkaline urine Protein reagent strip reaction . ´Protein Error of Indicatorsµ based on changing of indicator color in presence of changes in protein level instead of pH changes Tetra-bromphenol blue +protein >>>>>color change from yellow to green to blue green as protein increases Interference ² false positive due to highly buffered alkaline urine. highly pigmented urine. leaving urine on pad too long. detergent contamination Confirmation ² SSA test ² cold precipitation turbidity test.
hyperthyroidism .e.Glucose 100% of glucose normally reabsorbed in the proximal convoluted tubule Glycosuria occurs due to hyperglycemia (reaching the renal threshold 160 ² 180 mg/dl ² Diabetes mellitus ² Meal with high glucose content ² Hyperglycemia of non-diabetic origin i.
Glycosuria continued Glycosuria in the absence of hyperglycemia due to ´renal glycosuriaµ ² renal tubular reabsorption malfunction seen in end stage renal disease Normal blood glucose. positive urine glucose .
Glucose reagent strip reaction ´Double sequential enzyme reactionµ ² 1) glucose + O2 (glucose oxidase) >>>gluconic acid + H2O2 ² H2O2 + chromagen (peroxidase)>>>>oxidized colored chromagen ² Color change from green to brown as glucose increases .
Clinitest Can be used as a backup for glucose test strip Detects reducing substances (not only glucose) Copper reduction test ² old test was called Benedict·s test Used to check for galactosuria in newborns Inborn Error of Metabolism. 52 for comparisons of test strip glucose and Clinitest results . negative reagent strip for glucose but positive Clinitest Avoid ´pass throughµ with this test See chart on p.
Ketones Products of fat metabolism Appear in urine when carbohydrate utilization for energy is compromised due to: ² Diabetes mellitus ² insulin insufficiency ² Increased loss of CHO due to vomiting ² Inadequate intake of CHO ² starvation/malabsorption ² Strenuous exercise ² overuse of available CHO 3 Ketone bodies are beta-hydroxybutyric acid. acetoacetic acid and acetone .
Ketones evaporate in unpreserved urine. 65) Acetest tablet is used as a backup. See p.Ketone Reagent Strip reactions Acetoacetate + sodium nitroprusside in an alkaline medium >>>>purple color Test is very sensitive to acetoacetate and only slightly sensitive to acetone. can be used on serum or urine. Beta-hydroxybutyric and acetone are presumed present because acetoacetic acid is the intermediate compound and is easily converted to either of the two (see p. Lactose added gives better color differentiation. 54 for false pos reactions .
hemoglobinuria or myoglobinuria .Blood Hematuria ² intact RBCs. cloudy red urine Hemoglobinuria ² lysed RBCs. clear red urine Positive test strip for blood = hematuria.
glomerular disease. toxic chemicals. tumors.renal calculi. pylonephritis. anticoagulant therapy Non ² pathologic significance ² Strenuous exercise ² Menstrual contamination .Hematuria Pathologic significance ² Renal or genitourinary origin. trauma.
strenuous exercise . infections. Intravascular hemolysis ² No intact RBCs seen ² Hemolytic anemias.Hemoglobinuria Lysis of RBCs produced in UT. typically seen along with hematuria in UTIs. transfusion reactions. severe burns.
Hemoglobinuria vs Myoglobinuria ² Pink plasma ² Neg precip test ² ² . history very important ² Suspect in patients with muscle destruction. convulsions. history . trauma.Myoglobin Heme containing protein in muscle tissue ² Produces clear red/brown urine ² Pt. coma. LD enzymes .pos precip test .Pt.^ CK.clear plasma . muscle wasting diseases. etc.
Blood reagent strip reaction Pseudoperoxidase activity of hemoglobin H2O2 + tetramethylbenzidine in the presence of hemoglobin(peroxidase) >>>oxidized chromagen (green to blue) +H2O Hemoglobinuria >>>solid green to blue Hematuria >>> green speckled pattern See book for interfering substances .
protein reused by body ² Protoporphyrin converted to bilirubin Bilirubin transported to liver to be converted to water soluble compound (bilirubin glucaronide) Bilirubin glucaronide >>intestine>>uroblilinogen>>urobilin>>feces . protoporphyrin ² Iron. bile duct obstruction Urine appears burnt orange RBC breakdown: ² Iron. protein.Bilirubin Waste product of hemoglobin Seen in urine in liver disease.
Bilirubin reagent strip reaction Bilirubin glucaronide + diazo salt >>>> Azo dye (color change on strip ² tan/brown) Hard to interpret ² susceptible to urine pigments Ictotest ² used to confirm ² more sensitive and interfering substances are washed away .
gives feces brown color As it circulates in blood back to liver. so normal to have small amount of this in urine Abnormally high amounts (> 1mg/dl or Ehrlich unit) seen in urine in liver disease and hemolytic disorders Multistix 10 SG uses Ehrlich·s aldehyde reagent to produce red to pink color on reagent strip Watson-Schwartz test differentiates porphobilinogen from urobilinogen ² . small portion is filtered by kidney. 68 Most excreted in feces.Urobilinogen Produced from reduction of bilirubin by intestinal bacteria ² see chart p.
Nitrite Screening test for bacteriuria Positive in cystitis and pyelonephritis Most UTI·s caused from Gram negative bacilli that reduce nitrate to nitrite Also used to monitor success of antibiotic therapy and to screen people with recurrent infections Correlate with Leukocytes on test strip .
61 for false neg and false pos reactions .Nitrite test strip reaction Greiss reaction Nitrite + aromatic amine >> diazo compound + tetrahydrobenzoquinolin >> pink color Any color of pink = positive Will not react with bacteria that are non nitrate reducers (lack reductase) such as Staph and yeasts See p.
Leukocyte Test strip measures leukocyte esterase ² WBC need not be intact to give a positive result Used to screen for bacterial and non bacterial UTI Used to screen for inflammation of UT Correlates with Nitrite in order to determine necessity to do urine culture Does not quantitate # of leukocytes. this must be done by microscopic .
LE test strip reaction Leukocyte esterase + acid ester (on pad) >>>> aromatic compound +acid Aromatic compound + acid react with diazo salt >>>> purple azodye See book for reaction interference .
) Bromthymol blue pH indicator changes as pH changes More concentrated the urine>>more ions>>more H+>>more pH change>>color changes from blue to green to yellow .Specific Gravity Screens for the kidney·s ability to concentrate the urine (reabsorption capability) Reagent strip reaction based on dissociation constant (pK) change of a polyelectrolyte in an alkaline medium Polyelectrolyte ionizes (releasing H+ in relation to #of ions in solution.
005 to reading if urine is pH > 6.SG continued Measures only ionic solutes Eliminates interference by large molecules such as glucose and urea Elevated protein = slight increase due to protein anions Specimens with pH of >6.5 will have decreased readings due to interference with the indicator. Add .5 .
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