Primer and probe as well as kit used for avian influenza virus universal type direct RT-PCR detection

A technology of RT-PCR and avian influenza virus, which is applied in the field of primers, probes and kits for general-purpose direct RT-PCR detection of avian influenza virus, to achieve the effects of improving detection speed, saving costs and reducing operation steps

Inactive Publication Date: 2016-07-13
深圳澳东检验检测科技有限公司 +1
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Adopt primer and probe or test kit of the present invention, and in conjunction with other conventional reagent components, can realize in a PCR tube continuously, automatically carry out the release of virus RNA, reverse transcription and fluorescent quantitative PCR detection; From obtaining detection sample to It only takes 1.5 hours to obtain the test results; the sensitivity and accuracy are comparable to tradi

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 and probe as well as kit used for avian influenza virus universal type direct RT-PCR detection
  • Primer and probe as well as kit used for avian influenza virus universal type direct RT-PCR detection
  • Primer and probe as well as kit used for avian influenza virus universal type direct RT-PCR detection

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0031] In one embodiment of the present invention, the kit also includes a general-purpose internal standard solution of avian influenza virus; preferably, the above-mentioned internal standard solution is obtained by cloning the general-type M gene of avian influenza virus into the RNA pseudovirus expression vector pAR and obtaining The plasmid pAR-AIVUN was prepared by Escherichia coli transformation. Specifically, the preparation method includes:

[0032] (1) Cloning the universal M gene of avian influenza virus into the RNA pseudovirus expression vector pAR to obtain the plasmid pAR-AIVUN;

[0033] (2) Transform the plasmid pAR-AIVUN into Escherichia coli BL21;

[0034](3) The recombinant bacterium obtained in step (2) is cultured overnight in the LB liquid medium containing Amp;

[0035] (4) After taking the overnight culture and continuing to the logarithmic growth phase of the bacteria, IPTG induces the culture superfluid;

[0036] (5) collect the thalline, crudely e...

Embodiment 1

[0046] The RT-PCR detection method of the universal direct amplification of embodiment 1 bird flu virus

[0047] 1. Design of universal RT-PCR detection primers and probes for avian influenza virus

[0048] According to the nucleotide sequence of avian influenza in GenBank, the conserved sequence region of the M gene was selected as the amplified region, and the upstream and downstream primers of the general type of avian influenza virus and the detection probe of the general type of avian influenza virus were respectively designed by using the ABIPrimerExpress software. The sequences are as follows :

[0049] The sequence of the upstream primer (AIVUN-PF) of the universal type of avian influenza virus is:

[0050] 5'-TGAACACGACCTAGACTGACCCAGAA-3' (SEQ ID NO: 1);

[0051] The sequence of the downstream primer (AIVUN-PR) of the universal type of avian influenza virus is:

[0052] 5'-TGGCATGTTGGACACAATGTCAAG-3' (SEQ ID NO: 2);

[0053] The sequence of the universal detection...

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

No PUM Login to view more

Abstract

The invention discloses a primer and a probe as well as a kit used for avian influenza virus universal type direct RT-PCR detection. The primer and the probe comprise a detecting primer and a detecting probe of the avian influenza virus as well as a probe of internal label RNA. With the adoption of the primer and the probe, a liquid sample can be directly added into a reaction system, so that release, reverse transcription and fluorescent quantitative PCR detection of the virus RNA are continuously and automatically realized in one PCR tube; the purposes of extracting the virus RNA and directly detecting the avian influenza virus in the extracted sample are realized, so that only 1.5 hours are needed for obtaining detected results from a to-be-detected sample; and the defect that RNA is needed to be extracted by conventional RT-PCR is effectively overcome, operation steps can be greatly reduced, the detection speed is greatly increased, the cost is reduced, and cross contamination is effectively avoided.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a primer, a probe and a kit for general direct RT-PCR detection of avian influenza virus. Background technique [0002] Avian influenza (Avian Influenza, AI), also known as European chicken plague or true chicken plague, is an acute and highly contagious infectious disease caused by type A influenza virus. It is an internationally recognized devastating disease and can be transmitted to humans by pigs or other animals. Therefore, it has very important ecological and public health significance, especially highly pathogenic avian influenza is listed as a class A infectious disease by the World Organization for Animal Health. [0003] In recent years, it has been found that avian influenza not only seriously harms poultry but also mammals, especially can infect and cause death of humans, which greatly threatens the safety of human public health. The avian influenza virus is...

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): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/166C12Q2563/107C12Q2545/101
Inventor 翟建新孙彦伟卢受昇林琳潘艳萍吴晓卫高慧敏邓国栋余希尧张利
Owner 深圳澳东检验检测科技有限公司
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