Rabies virus NP-ELISA antibody detection reagent kit

Abstract

The invention relates to a dog rabies virus NP-ELISA antibody detection kit, wherein, an antibody detection plate is a detachable 96-hole enzyme labeled plate which is coated by rabies virus nucleoprotein NP, enzyme conjugate working liquid is a horseradish peroxidase labeled anti-dog IgG monoclonal antibody, a positive control is dog rabies standard positive serum, a negative control is a dog standard negative serum, the rabies virus nucleoprotein NP is a fragment of rabies virus ERA vaccine strain NP gene 1350bp which is amplified by RT-PCR technology, the rabies virus nucleoprotein NP is inserted into the downstream of a prokaryotic expression vector pET-28a to construct a pET-NP recombinant expression plasmid, Escherichia coli BL21 DE3 is converted by the calcium chloride method, 0.01mM IPTG induced expression is carried out, ultrasonic waves are used for the lysis of an expression strain, and the purification by a Ni-NTA affinity chromatography column is carried out for the preparation. The using concentration of the rabies virus nuclear NP recombinant protein for coating the NP antibody detection plate is 2 mug / ml, and the using amount is 100 mu 1 / hole. The dog rabies virus NP-ELISA antibody detection kit has the advantages of strong specificity, high sensitivity and good safety.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rapid diagnosis method and equipment for animal virus antibodies, in particular to a canine rabies virus NP-ELISA antibody detection kit. Background technique [0002] Rabies is an acute and highly fatal infectious disease caused by rabies virus, which mainly affects the central nervous system and causes acute, progressive and almost irreversible fatal encephalomyelitis. It is currently the most prevalent and harmful disease in the world. The worst zoonotic infectious disease. Rabies is one of the acute infectious diseases with the highest human fatality rate so far. There is no specific treatment drug, and the fatality rate is almost 100%. Rabies is mainly distributed in developing countries such as Asia, Africa and Latin America, except the United Kingdom, New Zealand, Japan and China Outside of Taiwan Province, other countries and regions have occurred. About 55,000 people die f...

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Problems solved by technology

[0003] At present, the methods used for the detection of rabies virus antigens and serum antibodies mainly include fluorescent antibody test (FAT), mouse neutralization test (MNT), rapid fluorescent focus inhibition test (RFFIT), fluorescent antibody virus neutralization test (FAVNT), Enzyme-linked immunosorbent assay (ELISA), etc., but these methods are whole virus antigens, which have certain risks in sample detection and evaluation, which limit their application to a certain extent

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