Recombinant expression plasmids used for packaging coxsackievirus B5 (CV-B5) pseudovirus, pseudovirus, kit and method

Abstract

The invention relates to recombinant expression plasmids used for packaging a coxsackievirus B5 (CV-B5) pseudovirus, the pseudovirus, a kit and a method. The recombinant expression plasmids used for packaging the coxsackievirus B5 pseudovirus are respectively named as the pEGFP-CV-B5 (417) plasmid and the pCVB3-replicon, the CV-B5 structural protein expressed by the pEGFP-CV-B5 (417) plasmid can be used for packaging CV-B3 subgenome RNA transcribed by the pCVB3-replicon in the cell, thus the CV-B5 pseudovirus is generated, the pseudovirus can be used for detecting the neutralizing antibody, and since the pseudovirus with single-cycle infection is adopted, the safety problem caused when the live virus is used is avoided. After a plurality of experiments, the result shows that the invention provides the method for detecting the CV-B5 neutralizing antibody which is safe, sensitive, rapid, specific, simple and convenient, and is low in cost. Based on the abovementioned features, the method is particularly suitable for the experiment for rapidly detecting the neutralizing antibody in large scale, and thus the method has the significant application value in developing viral vaccines and detecting the level of the CV-B5 specific neutralizing antibody of individual and group patients.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for detecting neutralizing antibodies of intestinal Coxsackievirus B5 and a special pseudovirus thereof, in particular to a recombinant expression plasmid for packaging Coxsackievirus B5 pseudovirus, a method for preparing the plasmid, Pseudovirus and rapid detection kit. Background technique [0002] Coxsackie virus (Cox V) is a serious pathogenic agent of human diseases, and its infection can cause many diseases, ranging from mild respiratory tract infection diseases to more serious myocarditis, pericarditis and some diseases of the nervous system , and can even cause infant death. The full length of the genome is 7389-7402 nucleotides. Coxsackieviruses can be divided into two groups, A and B. Group A has 24 types of viruses, and group B has 6 types of viruses. [0003] Coxsackievirus B5 type (Coxsakievirus B5, CV-B5) belongs to small ribonucleic acid virus (Picornavirida...

AI Technical Summary

Problems solved by technology

The antibody titer determined by the HI method has good reproducibility, and the VN method is superior to the HI method in sensitivity; another disadvantage of the HI method is that fresh blood cells must be used for each observation of agglutination.

In addition, not all virus neutralizing antibodies can inhibit hemagglutination, so this method is more suitable for use with virus neutralization tests, and is not suitable for large-scale neutralizing antibody screening tests

[0008] At present, methods for detecting neutralizing antibodies, such as live virus neutralization tests, mostly use completely infectious viruses. Although the results of such methods are reliable, the disadvantage is that live viruses are prone to antigenic drift during the passage process, and more importantly, There are dangers during the test operation, so there are high requirements for the operation and safety equipment of technicians

Other detection methods that do not require the use of live viruses also have various defects.

For example, the correlation with the results of the standard virus neutralization test is not good (ELISA method), the operation is cumbersome (antigen-antibody indirect agglutination inhibition test), and the cost is expensive (rapid fluorescent focus inhibition test), so it is not suitable for large-scale rapid screening

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