**ELISA Reagent Evaluation and Usage Guide**
Are you familiar with the evaluation of ELISA reagents? If not, Shanghai Hengyuan is here to help. We provide comprehensive support for all your ELISA-related needs. Feel free to visit our website or contact us directly for more information. Your satisfaction is our priority, and we look forward to your call.
The evaluation of ELISA reagents involves two main aspects: the quality of the reagent itself and its performance in clinical applications. For example, when evaluating hepatitis B ELISA diagnostic reagents, they must first pass verification by the Chinese pharmaceutical biological product authority to obtain a production license. In addition to proper packaging, labeling, and instructions, the reagent's performance—such as specificity, sensitivity, precision, and linearity—must be thoroughly tested using reference products. Only after passing these tests can the reagent be considered suitable for use.
In clinical settings, the quality of ELISA reagents is assessed based on their ability to distinguish between healthy and diseased states. The Central Inspection Center has conducted extensive work in this area, improving reagent quality through rigorous evaluations. This process typically involves collecting patient serum samples and testing them with the most reliable reagent available to determine whether they are positive or negative. While no test is 100% accurate, ELISA remains one of the most widely used methods due to its reliability and efficiency.
**Key Points for Clinical Quality Evaluation**
- The reagent’s reliability is evaluated from a clinical perspective, focusing on its ability to detect disease markers accurately.
- Specificity, sensitivity, precision, and linearity are critical factors that must be tested.
- A “seed plate†of known positive and negative sera is often used to validate the test results.
- No test is perfect, and false positives or negatives may still occur, but careful validation helps minimize errors.
**Kit Instructions and Handling**
1. Store the reagents away from strong light and keep all vials tightly sealed to prevent evaporation and contamination.
2. Handle the reagents carefully, following the recommended incubation times and temperatures. Avoid long time gaps between adding samples to different wells, as this can affect accuracy.
3. Washing steps are crucial; incomplete washing may lead to inaccurate results.
4. Some reagents should be stored at -20°C, while others require refrigeration at 2–8°C. Always refer to the label for storage conditions.
5. If the concentrated washing solution becomes cloudy, it can be dissolved by heating in a water bath.
6. Small droplets inside the microplate wells are normal and do not affect results.
7. There may be slight differences between Chinese and English manuals. Please follow the English version for accuracy.
8. All samples must be handled according to standard operating procedures.
9. Shelf life: 6 months.
**Sample Preparation and Requirements**
1. **Serum**: Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect the supernatant and avoid any cellular debris.
2. **Plasma**: Use EDTA or sodium citrate as an anticoagulant. Mix well and centrifuge for 20 minutes. Collect the supernatant carefully.
3. **Urine**: Collect in a sterile container and centrifuge for 20 minutes. Discard any precipitate.
4. **Cell Culture Supernatant**: Collect in a sterile tube, centrifuge, and carefully collect the supernatant. For intracellular components, lyse the cells using repeated freeze-thaw cycles before centrifuging again.
5. **Tissue Specimens**: Weigh the tissue, add PBS (pH 7.4), homogenize, and centrifuge. Keep the supernatant for testing and store the rest at 2–8°C.
6. **General Sample Handling**: Process samples as soon as possible after collection. If not used immediately, store at -20°C, avoiding repeated freezing and thawing.
7. **Avoid NaN3**: Samples containing sodium azide (NaN3) should not be used, as it inhibits horseradish peroxidase (HRP) activity.
For more details or assistance, please contact our sales team at Shanghai Hengyuan. We offer professional guidance and support to ensure the best results with our ELISA kits. Let us help you achieve accurate and reliable test outcomes.
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