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A rapid detection method for varicella-zoster virus titer

A technology of herpes zoster virus and virus titer, which is applied in the biological field, can solve the problems of affecting test results, long test cycle, and large impact of virus titer, and achieve the effect of shortening the time

Active Publication Date: 2020-04-14
CHANGCHUN KEYGEN BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method also has shortcomings. For example, the detection period is long and often takes more than a week. Secondly, the detection of virus titer is greatly affected by the cell matrix. When the cell state is not good, it greatly affects the detection results.

Method used

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  • A rapid detection method for varicella-zoster virus titer
  • A rapid detection method for varicella-zoster virus titer
  • A rapid detection method for varicella-zoster virus titer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: VZV double antibody sandwich ELISA detection kit

[0029] The VZV double antibody sandwich ELISA detection kit can be selected from the existing VZV double antibody sandwich ELISA detection kit, more preferably, the kit includes: ELISA detection plate coated with anti-VZV monoclonal antibody mAb-11, HRP enzyme labeling Anti-VZV monoclonal antibody 12-HRP, VZV antigen standard, known virus titer VZV vaccine standard, TMB chromogenic solution.

[0030] The present inventor screened hybridomas secreting VZV virus-specific monoclonal antibodies, obtained anti-VZV monoclonal antibodies mAb-11 and 12-HRP with excellent affinity and specificity, and determined the above-mentioned antibodies by competitive binding method. The affinity constant of the antibody, the results show that the affinity constant of mAb-11 is as high as 10 14 L / mol, the affinity constant of 12-HRP also reached 10 12 L / mol, both of them have very strong specific binding ability for VZV antige...

Embodiment 2

[0034] Embodiment 2: the mensuration of VZV vaccine virus titer

[0035] The present embodiment utilizes the test kit prepared in Example 1 to measure the VZV vaccine virus titer, and the specific assay method includes the following steps:

[0036] (1) Sample dilution:

[0037] VZV standard antigen dilution: Dilute the VZV standard antigen to 50ug / ml, 25ug / ml, 20ug / ml, 15ug / ml, 10ug / ml, 5ug / ml, 2.5ug / ml, 1ug / ml, 0.5ug with PBS solution / ml, 0ug / ml;

[0038] Dilution of vaccine standard with known titer: use PBS solution to make 2-fold serial dilutions of vaccine standard with known titer (5.5lg pfu / ml), every time the sample is diluted, the virus titer will be reduced on the original basis 0.3lg pfu / ml, diluted to a virus titer of 3.1lg pfu / ml.

[0039] (2) Sample addition:

[0040]Take the ELISA detection plate coated with the anti-VZV monoclonal antibody mAb-11 in the kit of Example 1 and equilibrate at room temperature; add the above-mentioned diluted sample and the sam...

Embodiment 3

[0066] Embodiment 3: the mensuration of VZV vaccine virus titer

[0067] In this embodiment, the kit prepared in Example 1 is used to measure the virus titers of four samples. The numbers of the four samples are respectively: 627-1, 627-2, 627-3, and 627-4. The specific measurement method includes the following steps :

[0068] (1) Sample dilution:

[0069] VZV standard antigen dilution: Dilute the VZV standard antigen to 50ug / ml, 25ug / ml, 10ug / ml, 5ug / ml, 2.5ug / ml, 1ug / ml, 0.5ug / ml, 0ug / ml with PBS solution;

[0070] Dilution of vaccine standard with known titer: use PBS solution to make 2-fold serial dilutions of vaccine standard with known titer (5.43lg pfu / ml), every time the sample is diluted, the virus titer will be reduced on the original basis 0.3lg pfu / ml, the virus was diluted to 3.03lg pfu / ml.

[0071] Subsequent (2) adding samples, (3) incubation, (4) washing the plate, (5) adding enzyme-labeled antibody (concentration is 0.05ug / ml), (6) washing the plate, (7) c...

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Abstract

The invention aims at providing a quick detection method of virus titer of varicella zoster virus (VZV). On the basis of traditional VZV antigen double-antibody sandwich ELISA detection method, the quantitative calibration is performed by adding the VZV sample with known titer, and the virus titer of the VZV is determined by drawing a VZV antigen content titer curve. The detection method is simpleand fast in operation, compared with the traditional pfu determination method, the detection efficiency of the VZV titer is greatly improved, and the method is especially suitable for the virus quickdetermination.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a rapid detection method for varicella-zoster virus titer. Background technique [0002] Varicella-zoster virus (varicella-zoster virus, VZV), belonging to the genus Human Herpesvirus, human herpesvirus type 3, its initial infection causes chickenpox, and then the virus stays latent in the ganglion, and reactivates when the body's immunity declines Causes shingles. [0003] At present, the main preventive method for chickenpox in China is vaccination, and the chickenpox vaccines are all attenuated live vaccines, while the herpes zoster vaccines marketed abroad are mainly high-titer live attenuated vaccines. Recently, the US FDA approved the gE subunit adjuvant vaccine produced by GlaxoSmithKline to prevent the occurrence of herpes zoster. At present, the titration of the live attenuated varicella-zoster vaccine is determined by the classic pfu method, which is currently the gold st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
CPCG01N33/56994
Inventor 朱晓文李春明沈红杰李海燕梁慧颖刘延威刘志强周慧赵海波王玮
Owner CHANGCHUN KEYGEN BIOLOGICAL PROD
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