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Mosquito-borne infectious disease pathogene premixed fluorescence PCR (polymerase chain reaction) detection reagent and kit

A technology for infectious diseases and pathogens, which is applied in the field of pre-mixed fluorescent PCR detection reagents and kits for pathogens of mosquito-borne infectious diseases, can solve the problems of complicated and tedious work in the preparation of detection reagents, poor stability of mixed reagents, operational errors and errors, etc. The effect of avoiding temporary liquid dispensing operation, stable PCR reaction, and avoiding errors

Inactive Publication Date: 2019-03-05
湖北省疾病预防控制中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, provide a kind of mosquito-borne infectious disease pathogen premixed fluorescent PCR detection reagent and kit, to solve the existing method for detecting mosquito-borne infectious disease pathogen can only be aimed at single pathogen, and The preparation of detection reagents is complex and tedious, takes a long time, and is prone to operational errors and errors; technical problems such as poor stability of mixed reagents

Method used

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  • Mosquito-borne infectious disease pathogene premixed fluorescence PCR (polymerase chain reaction) detection reagent and kit
  • Mosquito-borne infectious disease pathogene premixed fluorescence PCR (polymerase chain reaction) detection reagent and kit
  • Mosquito-borne infectious disease pathogene premixed fluorescence PCR (polymerase chain reaction) detection reagent and kit

Examples

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Embodiment 1

[0078] This embodiment provides a mosquito-borne infectious disease pathogen premixed fluorescent PCR detection reagent, which includes DEPC water, buffer solution without magnesium ions, magnesium chloride, dNTP, TTH enzyme, glycerol, bovine serum albumin and dimethyl sulfoxide As well as the dengue virus heterogeneous primers and probes, Zika virus heterogeneous primers and probes, yellow fever virus heterosexual primers and probes and chikungunya virus heterogeneous primers and probes, the concentration of each component is as follows in Table 4:

[0079] Table 4

[0080]

[0081]

Embodiment 2

[0083] This embodiment provides a mosquito-borne infectious disease pathogen premixed fluorescent PCR detection reagent, which includes DEPC water, buffer solution without magnesium ions, magnesium chloride, dNTP, TTH enzyme, glycerol, bovine serum albumin and dimethyl sulfoxide As well as the dengue virus heterogeneous primers and probes, Zika virus heterosexual primers and probes, yellow fever virus heterosexual primers and probes and chikungunya virus heterogeneous primers and probes, the concentration of each component is as follows in Table 5:

[0084] table 5

[0085]

[0086]

[0087] 2. Example of Premixed Fluorescent PCR Detection Kit for Mosquito-Borne Infectious Disease Pathogens

Embodiment 3

[0089] This embodiment provides a premixed fluorescent PCR detection kit for pathogens of mosquito-borne infectious diseases. The kit of this embodiment includes the following reagents:

[0090] (1) PCR reaction solution: the PCR reaction solution includes the mosquito-borne infectious disease pathogen premixed fluorescent PCR detection reagent in the above-mentioned embodiment 1;

[0091] Specifications: 8 tubes x 6 strips, 20μl / tube;

[0092] (2) Negative control

[0093] Specification: 1 tube, 40μl / tube

[0094] Ingredients: Tris-EDTA buffer (0.01M pH8.0);

[0095] Packaging: 0.6ml ordinary centrifuge tube with cover (purple transparent);

[0096] (3) Positive control

[0097] Specification: 1 tube, 40μl / tube

[0098] Components: Synthetic sequences of four mosquito-borne virus detection regions are transcribed into RNA in vitro, diluted with Tris-EDTA buffer (0.01M pH8.0) and stored frozen;

[0099] Packaging: 0.6ml ordinary centrifuge tube with lid (orange red tran...

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Abstract

The invention discloses a mosquito-borne infectious disease pathogene premixed fluorescence PCR (polymerase chain reaction) detection reagent and a kit containing the same. The mosquito-borne infectious disease pathogene premixed fluorescence PCR detection reagent comprises a mosquito-borne infectious disease viral specific primer and probe and base reaction fluid and reaction enzyme needed by fluorescence PCR, the base reaction fluid and reaction enzyme and the mosquito-borne infectious disease viral specific primer and probe are mixed together, so that a whole system of the mosquito-borne infectious disease pathogene premixed fluorescence PCR detection reagent can have premixed effects and keep stability in expiry date, the expiry date of six months or more is achieved, so that the reagent can be directly used in the PCR process, PCR stability is ensured, and detection results are accurate.

Description

technical field [0001] The invention belongs to the technical field of virus detection kits, and in particular relates to a mosquito-borne infectious disease pathogen premixed fluorescent PCR detection reagent and a kit. Background technique [0002] Mosquito-borne infectious diseases are natural foci of diseases transmitted by vector mosquitoes. Common ones include Japanese encephalitis, malaria, dengue fever, Zika, yellow fever, chikungunya fever and other highly harmful infections. sick. Mosquito-borne infectious diseases account for about 5%-10% of the total incidence of infectious diseases in my country every year, but the number of fatalities accounts for 30%-40% of the total number of infectious diseases. Emerging mosquito-borne infectious diseases are an important part of emerging infectious diseases, and they are intensifying globally. In 2014, a dengue fever virus outbreak broke out in China. As of 0:00 on October 21, 2014, a total of 38,753 dengue fever cases we...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12R1/93
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2563/107C12Q2537/143C12Q2521/101
Inventor 占建波田仁鹏窦文祥刘家发李阳占发先宋毅
Owner 湖北省疾病预防控制中心