Fluorescence quantitative PCR kit for detecting virus of aftosa and application

A foot-and-mouth disease virus and fluorescence quantitative technology are applied to a fluorescence quantitative PCR kit for detecting foot-and-mouth disease virus and its application field, and can solve the problem of inability to calculate the initial DNA or RNA copy number and the like

Inactive Publication Date: 2005-08-31
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since there is no linear relationship between the amount of final PCR product and the amount of initial template, the initial DNA or RNA copy number cannot be calculated based on the final PCR product amount

Method used

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  • Fluorescence quantitative PCR kit for detecting virus of aftosa and application
  • Fluorescence quantitative PCR kit for detecting virus of aftosa and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The composition and preparation of the fluorescent quantitative PCR kit of embodiment 1 foot-and-mouth disease virus and its reaction conditions:

[0031] A). Kit composition:

[0032] a) The composition of the kit is as follows: (10 reactions)

[0033] RNA extraction solution A (4ml / tube) RNA extraction solution B (800μl / tube)

[0034] One-step amplification reaction solution (315μl / tube) Reverse transcription and hot start Taq DNA polymerase (10μl / tube)

[0035] Strong positive standard (100 times diluted PCR reaction tube (sterile, RNase and DNase free)

[0036] Release standard product 20μl / tube, four tubes in total) DEPC H 2 O (2ml / tube)

[0037] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)

[0038] RNase inhibitor (400U / tube)

[0039] RNA extraction solution A is a product of Invitrogen Company, b) reverse transcriptase and hot start Taq DNA polymerase (2U / μl), c) one-step amplification (RT-PCR) reaction solution are all purchased f...

Embodiment 2

[0052] Embodiment 2 Fluorescence quantitative PCR kit detects the detection of different types of foot-and-mouth disease virus and compares it with plaque titration, a traditional method for quantitative determination of virus titer

[0053] A). Kit composition:

[0054] a) The composition of the kit is as follows: (10 reactions)

[0055] RNA extraction solution A (4ml / tube) RNA extraction solution B (800μl / tube)

[0056] One-step amplification reaction solution (315μl / tube) Reverse transcription and hot start Taq DNA polymerase (10μl / tube)

[0057] Strong positive standard (100 times diluted PCR reaction tube (sterile, RNase and DNase free)

[0058] Release standard product 20μl / tube, four tubes in total) DEPC H 2 O (2ml / tube)

[0059] Negative standard (50μl / tube) Borderline positive standard (50μl / tube)

[0060] RNase inhibitor (400U / tube)

[0061] RNA extraction solution A is a product of Invitrogen Company, b) reverse transcriptase and hot start Taq DNA polymerase (...

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Abstract

A fluorescence quantitative PCR reagent kit for detecting the foot-and-mouth disease virus features that the fluorescent probe (TaqMan) and the positive-sensor or antisence primer are used, and the in vitro transcripted RNA standard is directly used as the reference for both reverse transcription and amplification. It can be used for detecting the monitoring the foot-and-mouth disease and detecting the safety of animal and its food.

Description

technical field [0001] The invention relates to a fluorescent quantitative PCR kit for detecting foot-and-mouth disease virus, which is suitable for real-time monitoring of the outbreak of foot-and-mouth disease, can be widely used in the import and export of animal products and the safety detection of animal food, and can effectively and conveniently test the potency of virus vaccines. Carry out quantitative detection and quality control and provide technical support for related basic research, and also involve the use in the detection of foot-and-mouth disease outbreaks and virus vaccines. Background technique [0002] Foot-and-mouth disease virus is a small RNA virus that mainly causes foot-and-mouth disease (FMD) in artiodactyls. Its onset is characterized by rapid onset, rapid spread, high morbidity, and rapid decline in the production performance of sick animals, which has brought serious economic losses to animal husbandry and its internation...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/25C12Q1/68G01N33/569
Inventor 郑从义顾潮江屈三甫
Owner WUHAN UNIV
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