Precautions for using homocysteine assay kit

The use of homocysteine assay kit should pay special attention to the following precautions to ensure the accuracy and safety of the test results:

1. Sample collection and processing

Fasting blood collection

Fasting blood samples should be collected to avoid diet affecting homocysteine levels. After collection, it should be immediately refrigerated (2-8 ℃) and the measurement should be completed within 4 hours; If delayed detection is required, the serum should be refrigerated and sealed for storage (up to 48 hours at 2-8 ℃).

Avoid hemolysis and repeated freeze-thaw cycles

Hemolytic samples can interfere with the test results and should be avoided from use.

. If the sample needs to be stored for a long time, it should be packaged and frozen at -20 ℃ or -70 ℃, and repeated freezing and thawing should be avoided to prevent protein denaturation.

High concentration sample dilution

If the homocysteine content in the sample exceeds 50 μ mol/L, it needs to be diluted with physiological saline or 8.5% saline and measured. The result should be multiplied by the dilution factor.

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II. Storage and Use of Reagents

Storage Conditions

The reagent kit should be stored at 2-8 ℃ in the dark, and the shelf life is generally 1-2 years (refer to the instructions for details).

. The opened reagent can be stable for 30 days in the instrument compartment (2-8 ℃), but contamination should be avoided.

Preparation and Use of Reagents

Reagent 1 and Reagent 2 must be mixed according to the instructions, and different batches of reagents cannot be mixed.

. Enzyme labeled antibodies, biotin labeled antibodies, and other working fluids should be prepared before use to avoid loss of activity.

Avoid cross contamination

Use disposable tips to prevent the metal parts of the sampler from coming into contact with the reagent.

; The unused Flat noodles or reagents shall be sealed and stored.

III. Operating Norms

Instrument Calibration and Quality Control

Before testing, it is necessary to calibrate the instrument (such as fully automatic biochemical analyzer, enzyme-linked immunosorbent assay reader) to ensure the accuracy of parameters such as wavelength (such as 340nm, 450nm) and optical path (such as 1.0cm).

. Quality control is required for each batch of testing, and standard curves are drawn using standard samples to ensure that the results are within the reference range.

Sample addition and reaction conditions

Strictly control the sample amount (such as reagent 1185 μ L, reagent 250 μ L, sample 10 μ L) and reaction time (such as incubation at 37 ℃ for 5 minutes) according to the instructions.

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The enzyme cycle method requires a measurement temperature of 37 ℃, a wavelength of 340nm, a measurement mode of rate method, and a time of 120 seconds.

Plate washing and color development

The ELISA method requires thorough washing of the enzyme-linked immunosorbent assay plate (4-6 times) to avoid interference from unbound reagents.

. The color developing solution needs to be incubated in the dark for 30 minutes, and the detection should be completed within 15 minutes after adding the stop solution.

IV. Safety and Waste Management

Personal Protection

If reagents accidentally splash onto the skin or eyes, rinse immediately with clean water;

; Medical attention is required for accidental ingestion. Reagent 2 contains sodium azide, and rubber gloves should be worn during operation.

Waste disposal

All samples, detergents, and waste must be treated as infectious agents and comply with relevant laws and regulations.

. Avoid direct discharge of liquids containing chemical reagents to prevent environmental pollution.

V. Interpretation and Validation of Results

Comprehensive Analysis

The test results should be comprehensively judged based on the patient's medical history, symptoms, and other examination results (such as folate and vitamin B12 levels) to avoid single indicator diagnosis.

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Abnormal result review

If the result exceeds the reference range, it needs to be retested and confirmed, and checked for errors in sample processing, reagent preparation, and other processes.

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