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Method for detecting echovirus type 9 antibody

An antibody detection and virus technology, applied in the medical field, can solve the problems of high cost, labor and time, and achieve the effect of ensuring health

Pending Publication Date: 2022-04-15
杭州市疾病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The common disadvantages of these methods are time-consuming, laborious, high-throughput detection, high requirements for experimenters and equipment, high cost, and cannot be carried out in grassroots laboratories.

Method used

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  • Method for detecting echovirus type 9 antibody
  • Method for detecting echovirus type 9 antibody
  • Method for detecting echovirus type 9 antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0036] Embodiment: a kind of echovirus type 9 antibody detection method, such as figure 1 shown, proceed as follows:

[0037] S1: if figure 2 As shown, ECHO9 pET28a-VP1 recombinant plasmid preparation: ECHO9 VP1 sequence published in GenBank was optimized for rare codons in Escherichia coli, and the ECHO9 VP1 sequence was regenerated;

[0038] Plasmid pET28a+ expression reading frame designed amplification primers for VP1 gene:

[0039] Upstream primer: 5'-CATATGGGTAGGGTTGCTGACACAATACG-3', containing NdeI restriction site and initiation codon ATG;

[0040] Downstream primer: 5'-CTCGAGTTACACATACACAAGCTCCAGATTCTTGG-3', containing XhoI restriction site;

[0041] The newly synthesized ECHO9 VP1 sequence and the plasmid pET28a+ prokaryotic expression vector were double-digested with NdeI and XhoI, respectively. T4 ligase was used for ligation, and after ligation, the product was transferred to E.coli DH 5α for culture, and a single clone was picked for PCR, such as image 3 A...

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Abstract

The invention discloses a method for detecting an echovirus type 9 antibody, which takes ECHO9 VP1 recombinant protein as an antigen of an elisa plate, breaks through the limitation that a virus strain is directly inactivated to serve as an antigen to coat the elisa plate in the past or virus strain infected cells are used for preparing an antigen sheet, and meets the requirements of high throughput and safety in preparation of the elisa plate. The kit is beneficial to clinical treatment of sterile encephalitis diseases, and plays an important role in technical support for early discovery and early intervention of sudden epidemic situations caused by ECHO9, epidemiological research and guarantee of health of the masses.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a detection method for echovirus type 9 antibodies. Background technique [0002] Aseptic encephalitis outbreaks caused by Echovirus 9 (ECHO9) have been reported in the United States, Canada, the United Kingdom, Europe, Japan, South Korea and other places. In China, the incidence of viral encephalitis is relatively high, but reports of viral encephalitis caused by ECHO9 are less, which may be related to the lack of effective detection methods for ECHO9. At present, the etiological detection of ECHO9 mainly adopts virus culture, serum neutralization test or RT-PCR to amplify the target gene band, then conduct gene sequencing, and perform Blast comparison on the GeneBank website to determine the type of echovirus. The common shortcomings of these methods are time-consuming, laborious, high-throughput detection, high requirements for experimenters and equipment, high cost, and can...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543
Inventor 邱晓枫潘劲草汪皓秋
Owner 杭州市疾病预防控制中心
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