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D-Glucose + H2O + O2 ------------------------------- H2O2 + D-Gluconate
H2O2 + 4-AAP + Hydroxybenzoate---->Quinoneimine dye + H2O
Principle for Glucose Oxidase (equation)
Glucose Oxidase Principle
Glucose is oxidized by glucose oxidase to gluconate and hydrogen peroxide.
Phenol + 4-AAP + hydrogen peroxide, in the presence of peroxidase,
produces a quinoneimine dye that is measured at 500nm. The absorbance
at 500nm is proportional to the concentration of glucose in the sample.
Albumin is bound by the BCG dye to procedure an increase in the blue-green color measured at 630 nm. The color increase is proportional to the concentration of albumin present.
Alkali Protein + Cu++----------␣Colored Complex (protein conc.)
Total Protein (Biuret) Reagent Set Equation
Total Protein (Biuret) Principle
Protein in serum forms a violet colored complex when reacted with cupric ions in an alkaline solution. The intensity of the violet color is proportional to the amount of protein present when compared to a solution with known protein concentration.
Alkaline phosphatase principle
p-Nitrophenol phosphate is hydrolyzed to p-nitrophenol and inorganic phosphate. The rate at which the p-NPP is hydrolyzed, measured t 405 nm, is directly proportional to the alkaline phosphatase activity
Diagnosis: Glucose Oxidase Increase/Decrease
Blood sugar levels outside the normal range may be an indicator of a medical condition. A persistently high level is referred to as hyperglycemia; low levels are referred to as hypoglycemia. Diabetes mellitus is characterized by persistent hyperglycemia from any of several causes, and is the most prominent disease related to failure of blood sugar regulation.
Glucose Oxidase Normal Ranges
The normal range for Adults is 64.8 and 104.4 mg/dL. For children from birth to age 5, normal blood sugar targets are 100 to 200mg/dL. For children who are between the ages of 5 and 11, normal blood sugar targets are 70 to 150mg/dL.
What formula is used to calculate abs?
(Absorbance unknown sample /Concentration of Standard Absorbance) x concentration of standard =
Clinical Significance:Uric acid crystals are seen in normal urine. They may also be associated with kidney stone formation.
Clinical significance:Calcium oxalate crystals may be seen in normal urine. Calcium oxalate is the major component of kidney stones in adults
Clinical Significance:Calcium phosphate crystals are seen in normal urine. They may also be associated with kidney stone formation
Clinical Significance:Triple phosphate crystals are seen in normal urine. They may also be associated with kidney stone formation.
Clinical Significance:Ammonium biruate crystals can be seen in normal urine. However, the presence of ammonium biurate crystals usually indicates an old or poorly preserved specimen
Clinical significance:Cystine crystals are found in the inherited condition, cystinuria. Cystine crystals are the most frequent cause of kidney stones in children.
Clinical significance:Leucine crystals may be seen in liver disorders in which amino acid metabolism is impaired.
Clinical significance:Tyrosine crystals may be seen in tyrosinemia and in certain liver disorders in which amino acid metabolism is impaired.
Clinical Significance:Sulfonamide crystals may appear in the urine following antibiotic therapy with sulfonamides. They may be associated with kidney stone formation.
Clinical significance: WBC casts are associated with inflammation or infection within the kidney.
white cells present
The presence of RBCs in urine is clinically significant. They may be associated with disease, injury, or infection of any portion of the renal system.
Clinical Significance: Trichomonas is a common cause of vaginitis and is usually indicative of a STD.
Squamous epithelial cell
Clinical significance: Yeast is a common cause of urinary tract infection in immunocompromised or diabetic patients
Clinical Significance: RBC casts are the most diagnostic of all elements in urinary sediment. Their presence is always pathological. RBC casts are primarily indicative of an acute disorder of the glomerulus
Clinical significance: WBC casts are associated with inflammation or infection within the kidney. The presence of WBC casts is an indication that the WBC seen in the sediment are originating in the kidney
Clinical significance: Granular casts are occasionally seen in urine from normal individuals after strenuous exercise or stress. Increased numbers of granular casts may be seen in a variety of kidney diseases but are not indicative of any specific renal disorder.
Clinical Significance: Hyaline casts may be seen transiently following fever, stress, exercise or postural changes in normal individuals. Increased numbers of hyaline casts may be seen in all diseases of the kidney but are not indicative of any specific renal disorder.
Squamous epithelial cell*
Clinical Significance: Oval fat bodies are frequently associated with nephrotic syndrome.
oval fat bodies
The presence of RBCs in urine is clinically significant. They may be associated with disease, injury, or infection of any portion of the renal system
What is the proper storage of the reagent strips?
*protect from moisture/chemicals/heat and light
*keep lid tight and do not discard dessicant
*store below 30C (85F)
confirmatory test for bilirubin
place tablet on mat and add water
Diazo reaction=Bilirubin reacts w diazonium salt to produce tan, pink, violet
ability of glucose to reduce copper sulfate to cuprous oxide in presence of heat and alkali
contains all the reagents that are necessary for the reaction except the constituent to be measured.
Sample/ Serum blank
is useful when a serum is colored by a substance that are not being measured and do not interfere chemically with the reaction. ex)hemolyzed, lipemic or icteric samples
continuous movement in one direction; caused by reagent or instrument. (6 continuous results that are increasing or decreasing.
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