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Questions and Answers
How is the change in absorbance for the standard calculated?
How is the change in absorbance for the standard calculated?
What is the normal range for urea in serum/plasma?
What is the normal range for urea in serum/plasma?
What should be done if the urea concentration exceeds 300 mg/dl?
What should be done if the urea concentration exceeds 300 mg/dl?
Why is it necessary to run known controls with every assay?
Why is it necessary to run known controls with every assay?
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What is the recommended laboratory practice regarding the handling of reagents?
What is the recommended laboratory practice regarding the handling of reagents?
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What is the primary purpose of the LIQUIZYME UREA kit?
What is the primary purpose of the LIQUIZYME UREA kit?
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In the process described, which substance is NOT a product of the urea hydrolysis reaction?
In the process described, which substance is NOT a product of the urea hydrolysis reaction?
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Which of the following conditions is associated with elevated Urea levels?
Which of the following conditions is associated with elevated Urea levels?
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What is the wavelength used for measuring absorbance in the LIQUIZYME UREA procedure?
What is the wavelength used for measuring absorbance in the LIQUIZYME UREA procedure?
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What should be done with urine samples prior to analysis when using the LIQUIZYME UREA kit?
What should be done with urine samples prior to analysis when using the LIQUIZYME UREA kit?
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Which reagent is NOT included in the LIQUIZYME UREA kit?
Which reagent is NOT included in the LIQUIZYME UREA kit?
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How long is the working reagent stable when stored at 2-8 °C?
How long is the working reagent stable when stored at 2-8 °C?
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What is the main enzyme involved in the Urea hydrolysis reaction as per the LIQUIZYME UREA method?
What is the main enzyme involved in the Urea hydrolysis reaction as per the LIQUIZYME UREA method?
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Study Notes
Urea (UV) Test (UV GLDH Method)
- Intended Use: Quantitative determination of urea in serum, plasma, and urine.
- Clinical Significance: Urea is a waste product of protein metabolism. Elevated levels can indicate kidney disease, urinary obstruction, shock, or other conditions. Decreased levels might suggest liver failure or pregnancy.
- Principle: The urease enzyme hydrolyzes urea to ammonia and carbon dioxide. The ammonia reacts with ketoglutarate and NADH. The rate of NADH to NAD conversion is measured (as a decrease in absorbance) which is proportional to the urea concentration in the sample.
Procedure
- Reagents: Standard and sample require specific reagent volumes (1 ml, 10 μl).
- Reading: Measure the initial and final absorbance values of both standard and sample. Calculate the change in absorbance.
Calculation
- Formula: Specific formula for calculating urea concentration using the change in absorbance values (ΔAT/ΔAS)
Normal Values
- Serum/Plasma: 10-40 mg/dL
- Urine: 26-43 g/24 hours
Materials
- Clean glassware
- Laboratory pipette(s) or micropipettes
- Biochemical analyzer
Samples
- Serum, plasma, urine (avoid fluoride or ammonium ions in samples)
- Urine samples require dilution (urine 1: 49 with water, then multiply result by 50)
Reagent Preparation and Stability
- Mix Reagent 1 and Reagent 2 to create working reagent
- Stable for 5 days at room temperature (15-25°C)
- Stable for 4 weeks when stored at a temperature of 2 to 8.
- Opened kit stable til expiry
- Do not freeze reagents
System Parameters
- Reaction Type: Fix time (Decreasing)
- Wavelength: 340nm
- Cuvette Temperature: 37°C
- Delay Time: 30 Seconds
- Interval Time : 60 Seconds
Linearity
- Linear up to 300 mg/dL
- Sample dilution needed for values greater than 300 mg/dl
- Multiply result by dilution factor
Quality Control
- Run known controls regularly for accuracy
Limitations and Precautions
- Handle reagents carefully to avoid contact with skin, eyes, and mucous membranes
- Follow lab safety procedures for safe handling.
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Description
Test your knowledge on the Urea (UV) Test using the UV GLDH method. This quiz covers the intended use, clinical significance, principle, procedure, and calculations involved in determining urea concentration in biological samples. Perfect for students and professionals in clinical biochemistry!
