Primer pair, probe, kit and method for fluorescence quantitative RT-PCR detection of murine encephalomyocarditis viruses

A fluorescence quantitative, myocarditis technology, applied in the field of bioengineering, can solve the problems of cumbersome, low sensitivity and high price, and achieve the effect of optimizing reaction conditions, repeatability and specificity guarantee

Inactive Publication Date: 2017-06-09
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI +3
View PDF5 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used domestic and international detection method for mouse encephalomyocarditis virus (EMV) infection is serological ELISA detection, in which blood collection and serum separation of mice are particularly cumbersome and time-consuming, especially in large-scale experimental animal breeding institutions It is too cumbersome to use the traditional ELISA me

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primer pair, probe, kit and method for fluorescence quantitative RT-PCR detection of murine encephalomyocarditis viruses
  • Primer pair, probe, kit and method for fluorescence quantitative RT-PCR detection of murine encephalomyocarditis viruses
  • Primer pair, probe, kit and method for fluorescence quantitative RT-PCR detection of murine encephalomyocarditis viruses

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0028] Example 1 Fluorescence quantitative RT-PCR detection of murine encephalomyocarditis virus

[0029] 1. Experimental samples

[0030] 120 brain samples of mice from Shanghai Experimental Animal Research Center.

[0031] 2. cDNA synthesis

[0032] Take a small piece of mouse brain tissue, add PBS and steel balls into a shaker and shake for about 1 min, centrifuge at 4000 rpm for 2 min, aspirate the supernatant, and extract RNA. The extraction method refers to the instruction of TIANamp Virus RNA Kit extraction kit, and the extracted nucleic acid RNA is reverse transcribed to synthesize cDNA.

[0033] The template is extracted viral RNA, the primer pair is OligdT18, and cDNA is synthesized by reverse transcription. The reverse transcription system (40μL) is: RNA template 20μL, primer pair 2μL, 5×Reverse Transcription Buffer 8μL, dNTP (10mmol / L) 4μL, Ribonuclease Inhibitor (40U / μL) 1μL, Prime Script reverse transcriptase (200U) 1μL, DEPC water 4μL. 42℃ water bath for 1h, 70℃ water ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention discloses a primer pair, a probe, a kit and a method for fluorescence quantitative PCR detection of murine encephalomyocarditis viruses. The detection method takes cDNA of brain issues of a mouse as a template, SEQ ID No:1 and SEQ ID No:2 as primers and SEQ ID No:3 as the probe to perform fluorescence quantitative RT-PCR amplification and acquire a fluorescence signal. The fluorescence quantitative RT-PCR detection method disclosed by the invention can realize quick, accurate, convenient, rapid and specific detection of murine encephalomyocarditis virus nucleic acids in a mouse body, and the sensitivity, the repeatability and the specificity can be ensured.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and more specifically, the invention relates to a primer pair, a probe, a kit and a method for detecting mouse encephalomyocarditis virus by fluorescence quantitative RT-PCR. Background technique [0002] Murine encephalomyocarditis virus (EMV) is a member of the genus Cardiovirus in the family Picornaviridae and has only one serotype. Mice are the natural storage host of EMV, and EMV can infect a variety of host animals, including mammals, rodents, arthropods and humans, among which pigs are the most common and severe animal. Mice are extremely susceptible to EMV, and the main manifestations are acute encephalitis, myocarditis, diabetes, orchitis, and conjunctivitis. EMV is a single-strand positive-strand RNA virus with a genome length of about 7.8 kb and a large open reading frame (ORF). Its genome structure is similar to other picornaviruses. The genome homology of the porcine and mou...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2531/113C12Q2563/107
Inventor 陈鸿军魏晓锋蔡骁垚熊炜胡建华张泉
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap