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Preparation method for African swine fever virus antibody detection colloidal gold immunochromatography test paper strip

A technology of African swine fever virus and immunochromatographic test paper, which is applied in the field of biotechnology and animal quarantine detection, can solve the problems of no rapid diagnosis, and achieve the effect of simplifying sample preparation and shortening the detection time

Inactive Publication Date: 2015-04-08
YANGZHOU UNIV
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Problems solved by technology

Although many colloidal gold immunochromatographic test strips have been published at home and abroad, so far there is no attempt to use colloidal gold immunochromatographic technology for the rapid diagnosis of ASFV

Method used

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  • Preparation method for African swine fever virus antibody detection colloidal gold immunochromatography test paper strip
  • Preparation method for African swine fever virus antibody detection colloidal gold immunochromatography test paper strip
  • Preparation method for African swine fever virus antibody detection colloidal gold immunochromatography test paper strip

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Embodiment Construction

[0024] Source of biological material:

[0025] pET30a (+): Escherichia coli expression vector, introduced from Novogen, USA (Cat No.69909.3);

[0026] Escherichia coli (E.coli) BL21(DE3): imported from HaiGene Biotechnology Co., Ltd. (Cat No, K10127);

[0027] pGEX-4T-1-E183L: Contains the E183L (p54) gene of E70 strain ASFV, donated by Dr. RM Parkhouse of the Gulbenkian de Cie^ncia Institute in Portugal (Reis, A.L., R.M.Parkhouse, A.R.Penedos, C.Martins, and A.Leitao .2007.Systematic analysis of longitudinal serological responses of pigs infected experimentally with African swine fever virus.J.Gen.Virol.88:2426-2434.);

[0028] ASFV antibody positive and negative pig sera: provided by Dr. LK Linda, ASF Reference Laboratory, Pirbright Institute, UK (OIE ASFV Reference Laboratory).

[0029] The specific operation steps are as follows:

[0030] 1. Preparation of p54 recombinant antigen

[0031] The secondary structure of the amino acid sequence of E70 strain ASFV pE183L (p54...

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Abstract

The invention relates to a preparation method for an African swine fever virus (ASFV) antibody detection colloidal gold immunochromatography test paper strip, belonging to the field of bioengineering and animal epidemic disease diagnosis. The preparation method comprises the following steps of: expressing an ASFV recombinant antigen p54, purifying, preparing an antibody, preparing a colloidal gold immunochromatography test paper strip, and detecting the sensitivity and specificity of the ASFV antibody. Compared with the other conventional serological methods, the colloidal gold immunochromatography test paper strip prepared by the method has the advantages of convenience, quickness, sensitivity, specificity and the like, can be used for obtaining a definite diagnosis result within 5 minutes and directly detecting a suspicious swine serum (or anticoagulation) virus antibody, and is particularly suitable for field ASFV serological diagnosis, epidemiological survey and swine international trade quarantine and inspection.

Description

technical field [0001] The invention relates to the field of biotechnology and animal quarantine detection. Background technique [0002] African swine fever (ASF) is a highly fatal infectious disease of pigs. The disease has been prevalent in many African countries and has now spread to neighboring countries such as Georgia, Armenia, Azerbaijan and Russia. The threat is growing. At present, there is no vaccine for ASF, so rapid and accurate diagnosis is very important to prevent the introduction of the disease and its control. Diagnostic techniques for ASF recently recommended by the World Organization for Animal Health (OIE) include pathogen identification and serological tests. [0003] In pathogen identification technology, hemadsorption test is one of the classic ASF identification methods with high sensitivity, but it needs temporary preparation and culture of porcine bone marrow cells, which is not only time-consuming and cumbersome to operate, but also cannot be us...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
Inventor 张鑫宇孙怀昌夏晓莉
Owner YANGZHOU UNIV
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