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Chemical luminescence immune assay determination reagent kit for rubella virus IgM antibody and preparation method thereof

A chemiluminescence immunity and chemiluminescence technology, which is applied in the field of immunoanalysis medicine, can solve the problems of naked eye observation error, low accuracy rate, cumbersome measurement, etc., and achieve the effects of saving time and cost, improving accuracy and high sensitivity

Inactive Publication Date: 2009-02-18
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The technical effect of this patented technology lies between two methods: chemical reaction on an organic compound called lanthanide (L) particles, making it possible to create strong light signals from these particles without being affected by other substances like proteins found inside them; and binding certain molecules together into larger structures such as DNA strands through covalently crosslinked bonds, resulting in stronger signal intensity compared to direct measurement techniques. These improvements make the testing process faster and more efficient than existing tests.

Problems solved by technology

This patents describes various technical problem addressed in this patented text relating to the use of rubellaria vims (ruboled warts) due to its ability to spread through sexual contact between individuals without being affected. Current testing procedures involve collecting tissue specimens before laboratory treatment, followed up on hospital visits, and then subjective judgment based upon symptoms like abnormalities. However, existing tests lack precision and reliability, making accurate diagnocies difficult.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Preparation of rubella virus IgM antibody chemiluminescence immunoassay assay kit of the present invention

[0023] The rubella virus IgM antibody diagnostic kit of the present invention comprises:

[0024] Anti-human IgMμ chain-coated microwell plate (48-well or 96-well), negative and positive controls (1 bottle each), neutralizing antigen (1 bottle), alkaline phosphatase-labeled monoclonal antibody (1 bottle), sample diluent (1 bottle), concentrated washing solution (1 bottle) and chemiluminescence substrate (1 bottle).

[0025] 1. Preparation of rubella anti-human IgM μ chain-coated microwell plates

[0026] (1) Add the anti-human IgMμ chain to 0.02M PB buffer solution with a pH value of 4.2 and mix to make a coating solution with a concentration of 1:900. Add 100uL to each well in a microwell plate, and overnight at 37°C;

[0027] Specifically, the PB buffer preparation method is

[0028] Sodium dihydrogen phosphate 0.78g

[0029] Dilute purified water...

Embodiment 2

[0076] Embodiment 2 The usage method of the kit of the present invention

[0077] 1) Take out the kit from the refrigerator at 4°C, and equilibrate at room temperature for 15 minutes;

[0078] 2) Dilute the concentrated washing solution provided by the kit 20 times with deionized water;

[0079] 3) Adding samples: Set 3 wells for negative control and 2 wells for positive control, add 100uL of negative control and positive control respectively. Add 100uL of the sample diluent to the sample well, then add 10uL of the serum to be tested, set a blank well for each experiment, shake and mix well with a micro shaker, paste the sealing film, and incubate at 37°C for 30 minutes;

[0080] 4) Plate washing: Wash the plate 5 times with a fully automatic plate washer or manually, 400uL per well, and finally dry it on clean absorbent paper;

[0081] 5) Add neutralizing antigen and enzyme markers: except for blank wells, add 50uL each of neutralizing antigen and enzyme markers to each wel...

Embodiment 3

[0089] Embodiment 3 kit of the present invention is compared with ELISA kit

[0090] 1. Sensitivity:

[0091] Rubella IgM antibody-positive serum was tested after doubling dilution with the sample diluent, and the results are shown in Table 1:

[0092] Table 1 Sensitivity test results of two kits

[0093] serum titer 1:20 1:40 1:80 1:160 1:320 1:640 1:1280 1:2560 Kit of the present invention + + + + + + ± - ELISA kit + + + + ± - - -

[0094] It can be seen from Table 1 that when the dilution is 1:1280, the present invention can still be judged as positive, while the ELISA kit can only detect a dilution of 1:320, which shows that the present invention has higher sensitivity.

[0095] 2. Serum specimen testing:

[0096] The kit of the present invention and the ELISA kit simultaneously detect 487 cases of negative serum samples and 52 cases of rubella IgM positive serum samples that have been diagnosed, and the detection results o...

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Abstract

The invention provides a test kit and a preparing method thereof for rubella IgG antibody detecting by integrating chemiluminescence immunoassay with capture method. The test kit is composed of positive and negative reference substance, a micro-perforated plate which is coated by antihuman IgM u link, neutralizing antigen, sample diluent, condensed washing liquid, alkaline phosphatase labeled monoclonal antibody, and chemiluminescence zymolyte. The preparing method of the test kit includes: 1) the positive and negative reference substance is prepared; 2) the micro-perforated plate is coated by the antihuman IgM u link; 3) the neutralizing antigen is prepared; 4) the sample diluent is prepared; 5) the condensed washing liquid is prepared; 6) the rubella monoclonal antibody is labeled by the alkaline phosphatase; 7) the chemiluminescence zymolyte is prepared; 8) all components above can be filled and packed through separate packing and finished product can be made after assembly. The invention has the advantages that the sensitivity is high and the specificity is strong, thereby providing serology detection, epidemiology investigation and clinical diagnose with a reliable scientific diagnosing evidence which is helpful in eugenics.

Description

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Claims

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Application Information

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Owner CHEMCLIN DIAGNOSTICS CO LTD
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