Gold standard test paper for detecting antigens of peste des petits ruminants and preparation method thereof

A technology of Peste des petits ruminants and gold standard test paper, which is applied in the direction of biochemical equipment and methods, biological testing, chemical instruments and methods, etc., can solve the problems of shortage and poor conditions, and achieve the effect of low detection cost and simple preparation method

Inactive Publication Date: 2014-04-30
祁光宇
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Facing the reality of the poor conditions of the grass-roots testing institutions in my country and the lack of corresponding technical personnel, it has become a new diagnostic method that is fast, simple, relatively low in testing costs, does not require professionals, and can be operated on the testing site. Task

Method used

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  • Gold standard test paper for detecting antigens of peste des petits ruminants and preparation method thereof
  • Gold standard test paper for detecting antigens of peste des petits ruminants and preparation method thereof

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Embodiment Construction

[0011] The present invention will be explained below with reference to the embodiments.

[0012] The PPR N gene of the invention is an artificially synthesized sequence after gene optimization, wherein the optimization and artificial synthesis of the N protein gene, after the artificially synthesized N gene is connected with the pUC57-Simple cloning vector, the pUC57-Simple-N recombination is constructed Among the plasmids, the pUC57-Simple-N recombinant plasmid was constructed by Nanjing GenScript.

[0013] Peste des petits ruminants N of the present invention 1 -N 4 The gene is connected by bridge PCR method with the optimized sequence of N gene artificial synthesis, and the two pairs of ligation primers used are:

[0014] SEQ ID No. 1: PPRV-N 1 –F:CCG GAATTC ATGGCGACATTATTAA;

[0015] SEQ ID No. 2: PPRV-N 1 –R: CCGGAGGAACCACCCGCACCACGACTTGCGAAAGT;

[0016] SEQ ID No. 3: PPRV-N4-F: CGTGGTGCGGGTGGTTCCTCCG;

[0017] SEQ ID No. 4: PRV-N4-R: ATTT GCGGCCGC GCTCAGCAAGTCC...

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Abstract

The invention discloses gold standard test paper for detecting antigens of peste des petits ruminants and a preparation method thereof. The gold standard test paper contains a bottom plate, as well as a sample adding absorption pad, an immunogold standard pad constituted by a glass cellulose membrane adsorbing a gold standard antigen, a nitrocellulose membrane and a water absorption pad which are sequentially linked on the bottom plate, wherein a detection line and a quality control line are contained on the nitrocellulose membrane, the detection line on the immunogold standard pad is constituted by part of recombinant proteins N1-N4 of N genes after optimization of prokaryotic expression, and the quality control line is an anti-N1-N4 recombinant protein rabbit IgG antibody. The gold standard test paper disclosed by the invention has the characteristics of fast speed, simplicity, convenience, intuition, sensitivity, accuracy and relatively low detection cost.

Description

technical field [0001] The invention relates to a test paper for detecting pathogens and a preparation method thereof. Specifically, the invention relates to a gold standard test paper for detecting Peste des petits ruminants antigen and a preparation method thereof. Background technique [0002] Peste des Petits Ruminants is caused by Peste des Petits Ruminants virus (PPRV) of the Paramyxoviridae family Morbolivirus, which mainly infects small ruminants, especially goats and sheep. It is a severe, contagious disease. The World Food and Agriculture Organization (FAO) and the World Organization for Animal Health (OIE) list Peste des Petits Ruminants as a legally notifiable animal disease, and my country classifies it as a Class I animal disease. (Long Yunfeng, Liu Xiaohui, Zhou Xiaoli, et al. Research progress on the epidemiology and control of Peste des petits ruminants [J]. Advances in Veterinary Medicine, 2012, 33(5): 94-98.). PPRV was first discovered in Côte d'Ivoire, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C12N15/70C12N15/66
CPCG01N33/56983C07K14/005C12N15/70C12N2760/18422G01N33/558G01N2333/12
Inventor 祁光宇蒋韬刘斌王超英牟克斌刘学荣杜平董金杰智晓莹武发菊
Owner 祁光宇
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