Human Rotavirus IgG (RV-IgG) Enzyme Linked Immunoassay Kit Instructions for Use

This kit is for research use only. 48T
Drug Name:
Generic Name: Human Rotavirus IgG (RV-IgG) Enzyme-Linked Immunoassay Kit Uses:
This kit qualitatively measures rotavirus IgG (RV-IgG) in human blood, or other related tissues.
Experimental principle:
The kit uses double antibody sandwich enzyme-linked immunosorbent assay (ELISA) to determine human rotavirus IgG (RV-IgG) in the specimen.
The microplate is coated with purified rotavirus IgG (RV-IgG) antibody to prepare a solid phase antibody, which can be used with rotavirus in the sample.
The IgG (RV-IgG) antigen is combined, washed to remove unbound antigen and other components, and then combined with HRP-labeled rotavirus IgG (RV-IgG) antibody to form an antibody-antigen-enzyme-labeled antibody complex. Thoroughly wash and add substrate TMB
Color development. TMB is converted to blue under the catalysis of HRP enzyme and converted to the final yellow color by the action of an acid. The absorbance (OD value) was measured at 450 nm using a microplate reader and compared with the CUTOFF value to determine the human rotavirus in the specimen.
The presence or absence of IgG (RV-IgG).
Kit composition:
1 20 times concentrated washing solution 20ml × 1 bottle 7 stop solution 3ml × 1 bottle
2 enzyme standard reagent 3ml × 1 bottle 8 positive control 0.5ml × 1 bottle
3 enzyme label coating plate 12 holes × 4 strips 9 negative control 0.5 ml × 1 bottle
4 sample dilution 3ml × 1 bottle 10 instructions 1 copy
5 developer A liquid 3ml × 1 bottle 11 sealing film 2 sheets
6 developer B liquid 3ml × 1 bottle 12 sealed bag 1 specimen requirements:
1. Specimen processing: serum and plasma samples can be directly detected
2. The specimens were tested as soon as possible after preparation. If it cannot be detected in time, the specimen can be stored at -20 °C for one month.
However, repeated freezing and thawing should be avoided.
3. Samples containing NaN3 could not be detected because NaN3 inhibited horseradish peroxidase (HRP) activity.
Steps:
1. No.: The corresponding micropores of the sample are numbered sequentially. Each plate should have 2 holes for negative control, 2 holes for positive control, and blank control 1
Hole (the blank control well is not added with the sample and the enzyme standard reagent, the other steps are the same)
2. Loading: 50 μl of negative control and positive control (standard) were added to the negative and positive control wells, respectively. Then, add 40 μl of the sample dilution to the sample well to be tested, and then add 10 μl of the sample to be tested. Add the sample to the bottom of the well of the ELISA plate.
Try not to touch the hole wall, gently shake and mix,
3. Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes.
4. Dosing: Add 30 times concentrated washing solution to distilled water to 600ml.
5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing solution, let stand for 30 seconds, then discard it.

Repeat 5 times and pat dry.
6. Add enzyme: Add 50 μl of enzyme labeling reagent to each well, except for blank wells.
7. Incubation: The operation is the same as 3.
8. Wash: Operate the same as 5.
9. Color development: Add 50 μl of developer A to each well, then add 50 μl of developer B, gently shake and mix, and avoid light at 37 °C.
15 minutes
10. Termination: 50 μl of stop solution was added to each well to stop the reaction (the blue color turned yellow).
11. Measurement: The absorbance (OD value) of each well was measured sequentially with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution.
The result is judged:
Test validity: mean value of positive control well ≥ 1.00; average value of negative control ≤ 0.15
CUT OFF calculation: critical value = negative control well average + 0.15
Negative judgment: The sample OD value < CUT OFF is the rotavirus IgG (RV-IgG) negative positive judgment: the sample OD value ≥ the critical value (CUT OFF) is rotavirus IgG (RV-IgG) positive Precautions
1. The operation is carried out in strict accordance with the instructions. The components of the different batches of this reagent shall not be mixed.
2. The kit should be taken out from the refrigerated environment and allowed to equilibrate for 15-30 minutes at room temperature. If the enzyme label is unsealed after unsealing, the strip should be stored in a sealed bag.
3. Concentrated washing liquid may crystallize out. When diluted, it can be heated and dissolved in a water bath. The washing will not affect the result.
4. The sealing film is intended for single use only to avoid cross-contamination.
5. Please keep the substrate away from light.
6. The test results must be determined by the microplate reader. When using dual-wavelength detection, the reference wavelength is 630nm.
7. All samples, washings and various wastes should be treated as infectious materials. The stop solution is 2M sulfuric acid and must be used safely.
Storage conditions and expiration date
1. The kit is stored at: 2-8 °C.
2. Validity: 6 months

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