Method for separating, enriching and detecting enveloped viruses on basis of CL7-CVN and Im7 system

A CL7-CVN and capsule technology, applied in the field of biotechnology applications, can solve the problems of time-consuming, unfavorable rapid and on-site detection, complicated operation, etc., and achieve the best effect, excellent microstructure and swelling performance

Active Publication Date: 2020-05-29
HUBEI UNIV
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Problems solved by technology

There are many existing methods for the isolation and diagnosis of enveloped viruses, such as virus isolation and culture, PCR nucleic acid detection, enzyme-linked immunosorbent assay, etc.; however, these methods are complex in operation, time-consuming, high in technical requirements and expensive, and are not conducive to rapid and on-site testing

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  • Method for separating, enriching and detecting enveloped viruses on basis of CL7-CVN and Im7 system
  • Method for separating, enriching and detecting enveloped viruses on basis of CL7-CVN and Im7 system
  • Method for separating, enriching and detecting enveloped viruses on basis of CL7-CVN and Im7 system

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[0032]In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below. Apparently, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0033] The main materials involved in the embodiments of the present invention are as follows:

[0034] Escherichia coli Rosetta (DE3) (Merck), Escherichia coli BL21 (DE3) (Merck), pET28a plasmid (Merck), pET23a plasmid (Merck). 3C protease and PfuDNA polymerase were expressed and purified by our laboratory, T5 exonuclease was purchased from NEB Company, ultra-low molecular weight protein molecular weight standard protein was purchased from Boster Company, and primers were purchased from Sh...

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Abstract

The invention discloses a method for separating, enriching and detecting enveloped viruses on the basis of a CL7-CVN and Im7 system. The method for separating, enriching and detecting the enveloped viruses on the basis of the CL7-CVN and Im7 system comprises the following steps: firstly, producing recombinant CL7-CVN protein and Im7 protein; coupling the Im7 protein with beads to obtain beads Im7-Beads; putting the recombinant CL7-CVN protein into a to-be-enriched virus sample, after incubation is conducted, putting the Im7-Beads, conducting enrichment after incubation is conducted, and conducting centrifugation to obtain virus-enriched Im7-Beads; and extracting nucleic acid of the enveloped viruses enriched on the Im7-Beads, conducting RT-PCR detection, and calculating the enrichment efficiency. According to the method for separating, enriching and detecting the enveloped viruses on the basis of the CL7-CVN and Im7 system, CL7-CVN protein is produced through a genetic engineering method, and by means of specific binding of CL7 and Im7 and the performance of CVN of binding the enveloped viruses, surfaces of the Im7-Beads can be enriched with PRV virus particles, so that a brand novel technical measure is provided for separation, enrichment and detection of viruses.

Description

technical field [0001] The invention belongs to the application field of biotechnology and the field of environmental monitoring technology, and specifically relates to a method for separating, enriching and detecting enveloped viruses based on CL7-CVN and Im7 systems. Background technique [0002] At present, enveloped viruses such as HIV, herpes simplex virus, hepatitis C virus, influenza virus, pseudorabies virus, African swine fever, and Ebola virus have created a huge threat to human survival and development, and even disasters. There are many existing methods for the isolation and diagnosis of enveloped viruses, such as virus isolation and culture, PCR nucleic acid detection, enzyme-linked immunosorbent assay, etc.; however, these methods are complex in operation, time-consuming, high in technical requirements and expensive, and are not conducive to rapid And on-site testing and other shortcomings. Therefore, the establishment of a simple, rapid, accurate, and high-th...

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Application Information

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IPC IPC(8): C12N7/00C12N7/02C12Q1/70C12Q1/6851
CPCC12N7/00C12Q1/701C12Q1/6851C12N2710/16751C12Q2531/113C12Q2563/107C12Q2537/16
Inventor 余晓岚王斌王飞马立新杨智刘敏高丹陈冠军
Owner HUBEI UNIV
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