Podcast
Questions and Answers
What substance is formed when hydrochloric acid acts on fructose during Seliwanoff’s Test?
What substance is formed when hydrochloric acid acts on fructose during Seliwanoff’s Test?
What color indicates a positive result for fructose in Seliwanoff’s Test?
What color indicates a positive result for fructose in Seliwanoff’s Test?
What does the presence of sugar in urine typically indicate?
What does the presence of sugar in urine typically indicate?
Which of the following substances is NOT classified as a ketone body?
Which of the following substances is NOT classified as a ketone body?
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What happens to excess acetyl CoA in the liver?
What happens to excess acetyl CoA in the liver?
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Which cells primarily use ketone bodies as an energy source?
Which cells primarily use ketone bodies as an energy source?
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How are free fatty acids transported to the liver?
How are free fatty acids transported to the liver?
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Why can't the liver use ketone bodies as fuels?
Why can't the liver use ketone bodies as fuels?
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What is the first step in performing Esbach’s test for the total volume of urine?
What is the first step in performing Esbach’s test for the total volume of urine?
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When is it necessary to dilute urine before protein testing using Esbach's test?
When is it necessary to dilute urine before protein testing using Esbach's test?
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What indicates a concentration of protein in the urine greater than 6-8 gm/l?
What indicates a concentration of protein in the urine greater than 6-8 gm/l?
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What adjustment needs to be made if the urine pH is not acidic during Esbach’s test?
What adjustment needs to be made if the urine pH is not acidic during Esbach’s test?
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What does the presence of leukocytes and bacteria in urine suggest?
What does the presence of leukocytes and bacteria in urine suggest?
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What characterizes Bence-Jones protein?
What characterizes Bence-Jones protein?
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At what temperature does Bence-Jones protein coagulate during the heat test?
At what temperature does Bence-Jones protein coagulate during the heat test?
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What should be done after adding Esbach’s reagent to the urine sample?
What should be done after adding Esbach’s reagent to the urine sample?
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What is a characteristic of functional proteinuria?
What is a characteristic of functional proteinuria?
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Which condition is associated with postural (orthostatic) proteinuria?
Which condition is associated with postural (orthostatic) proteinuria?
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What type of proteinuria is primarily due to kidney disease?
What type of proteinuria is primarily due to kidney disease?
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Which of the following is NOT a cause of pre-renal proteinuria?
Which of the following is NOT a cause of pre-renal proteinuria?
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What is one method used to test for urinary protein?
What is one method used to test for urinary protein?
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Which condition does NOT cause systemic disease or renal pathology proteinuria?
Which condition does NOT cause systemic disease or renal pathology proteinuria?
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What is accidental or false proteinuria?
What is accidental or false proteinuria?
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What color indicates a positive result on the Test-Tape after 30 seconds?
What color indicates a positive result on the Test-Tape after 30 seconds?
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Physiological proteinuria is typically characterized by which of the following?
Physiological proteinuria is typically characterized by which of the following?
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How is the reagent area of the Diastix strip utilized?
How is the reagent area of the Diastix strip utilized?
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What is the function of glucose oxidase in Diastix?
What is the function of glucose oxidase in Diastix?
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What is the expected result when glucose is present in Rubner’s Test?
What is the expected result when glucose is present in Rubner’s Test?
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What is a characteristic of the reagent area in Test-Tape?
What is a characteristic of the reagent area in Test-Tape?
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Which of the following conditions might lead to fructose appearing in urine?
Which of the following conditions might lead to fructose appearing in urine?
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What should be done at 30 seconds when using a Diastix strip?
What should be done at 30 seconds when using a Diastix strip?
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What happens to the indicator color on Test-Tape when it oxidizes?
What happens to the indicator color on Test-Tape when it oxidizes?
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What does a positive result in Robert's Test indicate?
What does a positive result in Robert's Test indicate?
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What reagent is used in Heller's Test?
What reagent is used in Heller's Test?
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How sensitive is the Sulphosalicylic Acid Test for detecting protein?
How sensitive is the Sulphosalicylic Acid Test for detecting protein?
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What is the main action that allows the Heat and Acetic Acid Test to work?
What is the main action that allows the Heat and Acetic Acid Test to work?
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In the procedure of Robert's Test, what indicates a positive outcome?
In the procedure of Robert's Test, what indicates a positive outcome?
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What happens during the first step of the Sulphosalicylic Acid Test?
What happens during the first step of the Sulphosalicylic Acid Test?
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Why is the lower portion of urine not heated in the Heat and Acetic Acid Test?
Why is the lower portion of urine not heated in the Heat and Acetic Acid Test?
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Which of the following tests can detect protein without forming a ring?
Which of the following tests can detect protein without forming a ring?
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Study Notes
Test-Tape for Glucose
- A screening test for glucose
- Utilizes the same principle as Clinistix
- Employs a different oxidation-reduction indicator and material for reagent impregnation
- Reagents are impregnated on a tear strip of special paper
- Indicator is yellow in its reduced form and green to blue in its oxidized form
Procedure
- Tear off approximately 1 and 1/2 inches of the strip
- Dip the strip into the urine specimen and immediately remove it
- Wait 30 seconds and observe the appearance of green color
- A yellow color indicates negative test
- A green color indicates positive test
Diastix Reagent Strip for Glucose
- A specific test for glucose based on glucose oxidase
- Chemical reaction is the same as Clinistix
- Uses a different chromogen system
- Reagent area contains glucose oxidase, peroxidase, blue background dye, and potassium iodide as the chromogen
Procedure
- Dip the reagent area of the strip into the specimen
- Compare the test area with the color chart after 10 seconds
- Compare the test area with the color chart after 30 seconds for a semi-quantitative result
Rubner’s Test
- Add 3 gms (excess) of lead acetate to 10 ml of undiluted urine
- Shake well and filter
- Boil the filtrate and add 1 ml of concentrated ammonium hydroxide
- Boil the mixture
- A precipitate forms, indicating a positive test
- Lactose: Red solution + Red precipitate
- Glucose: Red solution + Yellow precipitate
Fructose
- Found in urine after consuming fruits, honey, syrup, and jams
- Can be found in liver disease
- Found along with glucose in the urine of diabetics
Seliwanoff’s Test for Fructose
- Hydrochloric acid acts on fructose to form a furfuraldehyde derivative
- Derivative reacts with resorcinol to produce a red-colored compound
- Add a few drops of urine to 5 ml of reagent in a test tube
- Boil the mixture
- A red color within half a minute indicates a positive test
Clinical Significance of Urine Sugar
- Normally, urine does not contain enough sugar for a positive test
- Presence of sugar in urine indicates diabetes mellitus
- Used for screening, confirming diagnosis, and monitoring diabetic therapy
Ketones
- Also known as ketone bodies
- A group of three related substances:
- Acetone
- Acetoacetate (acetoacetic acid)
- β-hydroxybutyrate (β-hydroxybutyric acid)
- Formed in the liver from acetoacetate (oxidation product of amino acid, fatty acid)
- Used as an energy source for extrahepatic tissues like brain and muscle cells
Ketone Bodies as Alternative Fuel
- Normal products of fat metabolism
- Not detectable in the blood or urine of healthy individuals
- Fat is broken down in tissues to glycerol and fatty acids
- Free fatty acids are transported to the liver by plasma albumin and broken down to acetyl coenzyme A (acetyl Co-A)
- Excess acetyl CoA is diverted into ketone bodies
- Ketones are transported to peripheral tissues and oxidized by the TCA cycle
Increased Permeability of the Glomerulus
- Normally, the glomerular membrane is impermeable to protein molecules
- Damage to the glomerular membrane allows protein molecules to pass through
Decrease in Normal Reabsorption
- Disease conditions affect reabsorption in the tubules
Types of Proteinuria
- Classified based on etiology and mechanism
- Functional Proteinuria
- Systemic disease or renal pathology Proteinuria
Functional Proteinuria
- Not associated with renal damage
- Accidental or false proteinuria
- Occurs due to mixing of urine with proteinous fluids (pus, blood, vaginal discharge)
- Physiological proteinuria
- Less than 0.5 gm/24 hrs
- Associated with fever, exposure to heat or cold, emotional stress, and late pregnancy
- Postural (orthostatic) proteinuria
- Associated with upright position or sitting for a long time
- Intermittent proteinuria that disappears when lying down
Systemic Disease or Renal Pathology Proteinuria
- Not due to primary renal disease
- Fever and toxic conditions
- Venous congestion
- Renal hypoxia
- Hypertension
- Myxedema (hypothyroidism)
- Bence Jones protein
- Primarily kidney disease
- Glomerulonephritis
- Nephrotic syndrome (primary and secondary)
- Destructive parenchymal lesions (tumor, infection)
- Protein added to urine at a point farther down the urinary tract from the renal parenchyma
- Infection of the renal pelvis or ureter
- Cystitis
- Urethritis or prostatitis
- Contamination with vaginal secretions
Test for Urinary Protein
- Precipitation or turbidimetric test
- Principle: protein is precipitated or coagulated out of urine
- Tests include:
- Robert's Test (ring tests)
- Heller's Test (ring tests)
- Sulphosalicylic Acid Test (non-ring tests)
- Heat and Acetic Acid Test (non-ring tests)
Robert's Test
- Principle: protein is precipitated using concentrated Nitric acid (HNO3), forming a white compact ring
- Place 3-5 ml urine in a test tube
- Add 3 ml of Robert's reagent below the urine
- A white ring at the zone of contact indicates a positive test
- Read the ring within 3 minutes
- Report results based on the type of ring formed
Heller's Test
- Same principle as Robert's Test
- Heller's reagent is concentrated nitric acid
- Same procedure as Robert's test
Sulphosalicylic Acid Test
- Principle: Sulphosalicylic acid solution (20%) precipitates any protein in urine
- More sensitive and reliable than heat method
- Detects 5 to 10 mg/dl of protein in urine
- Take 2 ml of centrifuged (clear) urine in a test tube
- Add equal amount of Sulphosalicylic acid reagent
- Shake gently and let stand for 10 minutes
- Note the degree of turbidity against a dark background
- Grade and report as a non-ring test
- Interpretation: report based on turbidity
Heat and Acetic Acid Test
- Principle: protein precipitates by heat
- Fill a test tube with clear urine and heat the upper portion for 2 minutes
- Compare the turbidity with known standards
- If turbidity is present, it may be due to phosphates, carbonates, or protein
- Estimate the concentration of protein in the unknown urine
- If unknown urine contains more than 100mg/dl protein, dilute and repeat
Quantitative 24 Hour Protein Determinations
- Esbach’s Test
- Measure the total volume and filter some of the urine
- Perform qualitative protein test (Robert's or strip test)
- Make dilution based on qualitative test results
- Measure pH of urine and adjust if necessary
- Add pumice powder to the 0.5 mark of the Esbach's tube
- Add urine to the “U” mark
- Add Esbach’s reagent to the “R” mark
- Mix slowly by inversion 10 times
- Wait 30 minutes and read the highest point of the column
- The result is in grams per liter of protein in the urine
- Multiply by the dilution factor if diluted
- Calculate and record the g % and g / 24 hrs
Clinical Significance of Proteinuria
- Proteinuria: >6-8gm/l indicates renal damage
- A positive protein test may be correlated with casts
- Timely diagnosis and treatment are essential to prevent permanent kidney damage
- Presence of protein, leukocytes, and bacteria in urine suggests urinary tract infection
Bence - Jones Protein
- Abnormal low molecular weight globulin consisting of light chains of immunoglobulin
- Contains amino acids except methionine
- Found in urine of patients with multiple myeloma, a malignant disease of plasma cells affecting bone
- Incidence of Bence-Jones proteinuria in multiple myeloma is 50% to 80%
Heat test to screen Bence -Jones Protein
- Principle: Bence Jones protein coagulates when heated to 40 to 600C and partially/wholly re-dissolves on boiling
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Description
This quiz covers the various testing methods for glucose, including the test-tape and Diastix reagent strip. You'll explore the principles, procedures, and indicators used in these tests. Test your knowledge on how glucose is detected in urine samples.
