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Enterovirus 71 detection method and detection kit

An enterovirus and kit technology, applied in the field of molecular immunology, can solve problems such as time-consuming, high cost, and tediousness, and achieve the effects of accurate detection results, sensitive detection, and simple preparation methods

Active Publication Date: 2016-01-20
浙江艾格生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to solve the time-consuming, cumbersome, and high-cost defects of the EV71 detection method in the prior art, and provide a fast, efficient, accurate, sensitive, and low-cost EV71 virus detection method and its kit

Method used

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  • Enterovirus 71 detection method and detection kit
  • Enterovirus 71 detection method and detection kit
  • Enterovirus 71 detection method and detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Preparation of Polyclonal Antibody

[0031] 1.1 Preparation of immunogen

[0032] 1.1.1 Design of EV71VP1 gene sequence primers

[0033] According to the EV71 capsid protein VP1 gene sequence queried in GenBank (accession number: EU024958.1), primers were designed by ourselves. '-CCGCTCGAGGAGDGTGGTGATRGCRGTGCGACT-3'.

[0034] 1.1.2 RNA extraction of EV71 virus

[0035] Extract the RNA of EV71 virus (gifted by Zhejiang CDC) as a template:

[0036]Use the viral RNA extraction kit from Roche Company in the United States to directly extract RNA from the cultured virus. The operation steps are carried out according to the instructions in the kit. The method is as follows:

[0037] 1) Take a 1.5mL centrifuge tube, mark it and add 409μL of the working solution that has been mixed;

[0038] 2) Add 200 μL of the sample to be extracted with an RNase-free tip with a filter element, shake and mix on a vortex mixer, centrifuge briefly, and let stand at room temperature for 1...

Embodiment 3

[0088] 1.1 Use the prepared kit for sample detection

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Abstract

The invention discloses an enterovirus 71 double antibody sandwiched ELISA kit. The kit comprises an enzyme label plate coated with an anti-VP1 polyclonal antibody, an enzyme-labeled anti-VP1 polyclonal antibody and a VP1 antigen. The invention also discloses a preparation method and a detection method of the kit. The kit is prepared from the anti-VP1 polyclonal antibody and the enzyme-labeled anti-VP1 polyclonal antibody. The preparation method has simple processes. The kit realizes fast detection and standardization. Through use of high bonding titer of the anti-VP1 polyclonal antibody to various EV 71 genotypes, the detection result is more accurate.

Description

technical field [0001] The invention relates to the field of molecular immunology, in particular to a detection method and kit for enterovirus 71. Background technique [0002] Hand, foot and mouth disease (hand, foot and mouth disease; HFMD) is an acute infectious disease caused by enterovirus. Since the disease was first reported in 1957, it has been widespread for many times. In 2006, the World Health Organization (World Health Organization; WHO) listed this disease as the fourth leading cause of death. HFMD caused by various types of coxsackie, echovirus and enterovirus type 71 (EV71) often occurs in the United States, Australia, Italy, France, the Netherlands, Spain, Romania, Brazil, Canada, Germany and other countries. In 1998, an EV71 epidemic broke out in Taiwan, and more than 120,000 people were infected, and 78 people died. Epidemic of hand, foot and mouth disease in Singapore in 1998-2001 and a large-scale disease outbreak in China in 2007. Hand, foot and mouth...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/536G01N33/535G01N21/76
CPCG01N21/76G01N33/535G01N33/536G01N33/56983
Inventor 毕爱国刘诚毛慧慧
Owner 浙江艾格生物科技股份有限公司