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Enzyme immunological method for quickly detecting rubella virus antibody

A technology of rubella virus and enzyme immunity, which is applied in the field of clinical medical testing, can solve the problems of complex monoclonal antibody preparation process, high price, low sensitivity, etc., achieve clear reaction mode, good stability and repeatability, and prolong reaction time Effect

Inactive Publication Date: 2003-08-06
WUHAN UNIV
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AI Technical Summary

Problems solved by technology

[0033] Enzyme-linked capture method (EIA) is used to detect rubella virus IgM antibody, which has significantly improved specificity compared with conventional indirect ELISA method, and can eliminate the influence of RF factors that may appear in ELISA method, but this method is less sensitive than indirect ELISA method, and cumbersome steps
[0034] In addition, there is an indirect ELISA method using monoclonal antibodies as enzyme-labeled antibodies, which can also greatly improve the specificity and sensitivity of traditional methods. However, due to the complicated preparation process of monoclonal antibodies and the high price, it is not suitable for popularization.

Method used

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Experimental program
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Embodiment Construction

[0052] Serum samples: 39 positive sera were collected from patients with rubella in the acute phase of the outpatient clinic of the hospital, which were detected as positive for rubella virus IgM antibody by the kit (produced by Jingmei Bioengineering Co., Ltd.); 184 negative sera were collected from the physical examination serum of female students of childbearing age. It was tested negative by the above kit.

[0053] Implementation steps:

[0054] 1. Prepare carbonate buffer solution, dissolve 1.59 grams of sodium carbonate and 2.93 grams of sodium bicarbonate in 1000 milliliters (ml) of double distilled water.

[0055] 2. The rubella soluble antigen was diluted 1:400 with carbonate buffer and coated on a 96-well polystyrene reaction plate. Add 100 microliters (ul) to each well and incubate overnight at 4°C.

[0056] 3. Prepare phosphate buffer saline (PBS) with a pH value of 9.6, heat and dissolve 4.8 grams of disodium hydrogen phosphate in an appropriate amount of double ...

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PUM

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Abstract

The invention discloses an enzyme immunological method of rapidly detecting the antibody of the measles virus, mainly using PEG to speed up the antigen-antibody reaction, thus shortening the reactiontime of routine ELISA. Firstly, wrap the soluble antigen in the enzyme-mark board; secondly made up the PBS lotion, PH9.6, and the sample diluent containing PEG; thirdly use the sample diluent to dilute the measured blood serum and the positive and negative comparison blood serum, fourthy add the diluted blood serum into the hole of the enzyme-mark board for warm cultivation; fifthly throw off the antigen solution in the hole, and use PBS lotion to wash; sixthly use the sample diluent to dilute the known enzyme-mark antibody; seventhly add the diluted enzyme-mark antibody in the hole for warmcultivation.

Description

technical field [0001] The invention belongs to the field of clinical medical detection, and more specifically relates to an improved enzyme immunology method for detecting rubella virus. Background technique [0002] Rubella virus is one of two viruses that can cause severe deformities in fetuses. Primary infection of rubella virus in pregnant women in the first four months of pregnancy, whether dominant or recessive, can lead to miscarriage, stillbirth and congenital rubella syndrome. Therefore, early diagnosis of rubella virus infection is extremely important for prenatal and postnatal care and improving the quality of the birth population. At present, the early diagnosis of rubella mainly depends on the detection of specific antibody in serum, that is, immunoglobulin M (IgM). Conventional indirect enzyme-linked immunosorbent assay (ELISA) is widely used in China. Due to its cumbersome operation and long process, how to simplify the operation Steps to shorten the detect...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531G01N33/535G01N33/543G01N33/569
Inventor 周俊杨占秋徐连根肖红文莉
Owner WUHAN UNIV
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