Hand-foot-mouth disease detection reagent kit and its detection method

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: 2012-08-22
刘志学 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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 neutra

Method used

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  • Hand-foot-mouth disease detection reagent kit and its detection method
  • Hand-foot-mouth disease detection reagent kit and its detection method
  • Hand-foot-mouth disease detection reagent kit and its detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0032] The following examples further illustrate the invention, but should not be construed as limiting the invention.

[0033] The kit contains the following components:

[0034] (1) Viral RNA extraction reagent

[0035] Viral RNA can be extracted using the conventional Trizol method, or viral RNA extraction kits produced by various biological companies.

[0036] (2) LAMP detection reaction solution

[0037] ①Reaction solution Cox A16-used to detect Coxsackie virus A16

[0038] Contains the following ingredients:

[0039] 10x LAMP buffer (200mmol / L Tris-HCl, 100mmol / L KCl, 20mmol / LMgSO 4 , 100mmol / L (NH 4 ) SO 4 , 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.

[0040] Wherein the primer sequence is:

[0041] CoxA16-FIP:

[0042] 5-AAGTRGGTTTCGGAGCCCCTTTTTTAGCCAAACCCAATGGTGAG-3

[0043] CoxA16-BIP:

[0044] ...

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Abstract

The invention belongs to biological field, and relates to a reagent kit for detecting main pathogen Coxsackievirus A16 (Cox A16) and enterovirus 71 of hand-foot-mouth disease, construction method and uses thereof. The reagent kit comprises reaction solutions for detecting four primers of Coxsackievirus A16 and four primers of enterovirus 71.As detected, the reagent kit has a good performance of specificity, sensitivity, rapid amplification, high efficiency and simple identification. The detection system of the invention can detect Coxsackievirus A16 and enterovirus 71 quickly, conveniently, high-effectively, high-specifically and highly-sensitive under temperature of 64 DEG C without need of complicated instruments, which can preferably satisfy clinical detection for hand-foot-mouth disease and be easily popularized in large scope.

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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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 刘志学许馥慧
Owner 刘志学
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