Loop-mediated isothermal amplification assay kit and detection method of hand, foot and mouth disease

A detection kit and technology for hand, foot and mouth disease, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve the problems of time-consuming, laborious and cumbersome steps, and achieve simple operation methods, high sensitivity, and high accuracy Effect

Inactive Publication Date: 2011-11-16
上海吉美生物工程有限公司
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The traditional detection method of hand, foot and mouth disease pathogens includes two steps of virus isolation and neutralization test identification, which takes more than a week, and the steps are cumbersome, time-consuming and labor-intensive

Method used

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Embodiment Construction

[0043] The following examples further illustrate the invention, but should not be construed as limiting the invention.

[0044] The kit contains the following components:

[0045] (1) Viral RNA extraction reagent

[0046] Viral RNA can be extracted using the conventional Trizol method, or viral RNA extraction kits produced by various biological companies.

[0047] (2) LAMP detection reaction solution

[0048] ①Reaction solution Cox A16-used to detect Coxsackie virus A16

[0049] Contains the following ingredients:

[0050] 10 x LAMP buffer (200mmol / L Tris-HCl, 100mmol / L KCl, 20mmol / L MgSO 4 , 100mmol / L (NH 4 )SO4 , 1.0%Tritonx-100); 100mmol / L MgSO 4 25mmol / LdNTP; 5mol / L betaine (betaine); 20umol / L primer CoxA16-FIP; 20umol / L primer CoxA16-BIP; 5umol / L primer CoxA16-F3; 5umol / L primer CoxA16-B3; DEPC treated water.

[0051] The primer sequences are:

[0052] CoxA16-FIP:

[0053] 5-AAGTRGGTTTCGGAGCCCCTTTTTTAGCCAAACCCAATGGTGAG-3

[0054] CoxA16-BIP:

[0055] 5-TTYGC...

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Abstract

The invention belongs to the field of biotechnology, and relates to a loop-mediated isothermal amplification (LAMP) assay kit of Coxsackie A16 (Cox A16) and Enterovirus 71 (EV 71) which are main pathogens of hand, foot and mouth disease, and an establishment method and an application of the assay kit. The assay kit comprises four LAMP primers and LAMP reaction liquid for detecting Coxsackie A16, and four LAMP primers and LAMP reaction liquid for detecting Enterovirus 71. Tests prove that the assay kit has the advantages of good specificity and sensitivity, rapid amplification rapid, high efficiency and simple identification. A detection system provided by the invention can rapidly, conveniently, efficiently, high specifically and high sensitively detect Coxsackie A16 and Enterovirus 71 without complex apparatuses thus can satisfy well clinical detection requirements of hand, foot and mouth disease, and is suitable for a large-scale promotion and an application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for rapidly detecting the pathogen of hand, foot and mouth disease by using a loop-mediated isothermal amplification (LAMP) technology, and to a method for rapidly detecting the pathogen of hand, foot and mouth disease using the kit. Background technique [0002] Hand, foot and mouth disease (HFMD) is a global infectious disease caused by enterovirus. It has been reported in most parts of the world. Herpes can cause myocarditis, pulmonary edema, meningoencephalitis and other complications in a small number of patients. The main pathogens causing the disease are Coxsackie virus group A type 16 (Coxsackie A16, CAV16) and enterovirus type 71 (Enterovirus 71, EV71). The traditional detection method of HFMD pathogens includes two steps of virus isolation and neutralization test identification, which takes more than a week, and the steps are cumbersome, time-consuming and labor-intens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 刘代新刘军辉李昂
Owner 上海吉美生物工程有限公司
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