Biochemical test reagents play an important role in clinical diagnosis, scientific research, and industrial testing. Their standardized operation directly impacts the accuracy and reliability of test results. This article systematically describes the standard operating procedures for biochemical test reagents to ensure the scientific and reproducible nature of experimental data.
I. Pre-Operation Preparation
Environmental Requirements: Testing should be performed in a clean, well-ventilated laboratory, away from direct sunlight and temperature fluctuations (generally controlled between 18-25°C).
Reagents and Equipment: Before use, confirm that the reagents are within their expiration date and free of precipitation or discoloration. Supporting equipment (such as cuvettes, pipettes, centrifuges, etc.) must be calibrated and maintained sterile.
Sample Handling: Biological samples such as serum and plasma should be free of interference from hemolysis, lipemia, or icterus. Pre-treatment (e.g., centrifugation, dilution, etc.) should be performed according to the manufacturer's instructions.
II. Operational Procedures
Reagent Preparation: If the reagent is a lyophilized powder or concentrate, reconstitute or dilute it strictly according to the proportions specified in the manufacturer's instructions, mix thoroughly, and allow to cool to room temperature.
Sample Addition: Use a calibrated pipette to accurately pipette samples and reagents to avoid air bubbles. The order of sample addition generally follows "sample → reagent 1 → reagent 2" to ensure a uniform reaction system.
Reaction Condition Control: Place the reaction system in a constant-temperature water bath or spectrophotometer, setting the temperature (e.g., 37°C) and reaction time (usually 5-30 minutes) according to the reagent requirements.
Detection and Reading: Measure the absorbance (OD) using a spectrophotometer, selecting the wavelength specified for the reagent (e.g., 340nm, 546nm, etc.) to ensure that the instrument parameters match the reagent.
III. Precautions
Cross-contamination Prevention and Control: Replace pipette tips between samples or reagents to prevent residual contamination of the results.
Quality Control: Each batch of testing must include quality control samples. Results must be within the acceptable range; otherwise, retesting is required.
Waste Disposal: Waste reagents and samples should be disposed of separately according to biosafety regulations to prevent environmental contamination.
IV. Interpretation and Recording of Results
After the test is completed, calculate the results according to the standard curve or the formula in the reagent manual. When recording the data, please indicate the test time, instrument model, and operator information. Abnormal results should be considered in light of the clinical context or repeated verification.
Proper operation of biochemical test reagents is a key component in ensuring test quality. Operators must strictly follow standardized procedures and receive regular training to enhance their skills.