Peste des petits ruminants virus antibody detection kit based on ipma

A kind of PPR and kit technology, which can be used in microorganism-based methods, measuring devices, and cells modified by introducing foreign genetic material, etc., which can solve the problems of unconditional development, high workload and labor intensity, and expensive use. , to achieve the effect of intuitive interpretation

CN104991061BActive Publication Date: 2017-04-12HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2017-04-12

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Abstract

The invention provides a peste des petits ruminants virus antibody detection kit based on an immunoperoxidase monolayer assay (IPMA). The kit comprises a 96-well cell culture plate containing intracellular positive antigens and intracellular negative antigens, positive control serum, negative control serum, sample diluents, saline scrubbing solutions, HRP labeled mouse anti-goat McAb, two-component AEC colorimetric solutions, stop solutions, a serum diluting plate and an operating instruction. The intracellular positive antigens are BHK-gSLAM cells infected by a peste des petits ruminant virus rPPRV / GFP, the intracellular negative antigens are BHK-gSLAM cells not infected by the peste des petits ruminant virus, the positive control serum is immune positive serum obtained through blood sampling and separating after a goat is immunized by peste des petits ruminants virus vaccine strains PPRV N75 / 1, and the neutralizing titer is above 1:160; the negative control serum is negative serum obtained through direct blood sampling and separating from a non-immunized peste des petits ruminants negative goat.
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Description

technical field

[0001] The invention provides a detection kit of Peste des petits ruminants virus antibody based on immunoperoxidase cell monolayer assay (immunoperoxidasemonolayer assay, IPMA), and the kit can detect whether there are small ruminants in the body of small ruminants with high specificity and high sensitivity. Peste des petits ruminants virus (PPRV) antibody, so as to achieve the purpose of vaccine antibody detection or disease diagnosis. Background technique

[0002] Peste des petits ruminants virus (PPRV) infection is a severe and contagious infectious disease caused by peste des petits ruminants virus (PPRV), which can cause significant economic losses. The disease is a class A severe infectious disease stipulated by FAO / OIE, and a class I animal epidemic disease stipulated by my country.

[0003] Its pathogen is a member of the genus Morbillivirus in the Paramyxoviridae family, and its genome is a single-stranded negative-sense RNA that is not segmented. ...

Examples

Embodiment Construction

[0064] The present invention will be further described below with reference to specific examples, but those skilled in the art should understand that the present invention is not limited to these specific examples.

[0065] 1. Material method

[0066] 1.1 Cells and viruses The cells are BHK-21 cells (BHK-gSLAM) stably expressing goat SLAM protein (gSLAM). The virus is a PPRV N75 / 1 vaccine strain (rPPRV / GFP) carrying a green fluorescent protein (GFP) tag. All are preserved and provided by the important zoonoses and severe exotic diseases innovation team of Harbin Veterinary Research Institute (Yin, Chen et al.2014; see also patent application number: 201010559545.8).

[0067] 1.2 Serum 19 positive sera of Peste des petits ruminants (PPRV) with known virus neutralization test (VNT) titers were used for the sensitivity test of the IPMA method. 60 samples of Peste des petits ruminants negative sera (from sheep that were negative for Peste des petits ruminants, verified by VNT an...