Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

RT-LAMP (reverse transcription loop-mediated isothermal amplification) visual detection kit for H1N1 subtype avian influenza viruses

An avian influenza virus and kit technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of high cost, long time, and unsuitable for grass-roots promotion and use.

Active Publication Date: 2012-08-29
GUANGXI VETERINARY RES INST
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional chicken embryo isolation method of virus is accurate and simple, but it takes too long, usually takes two days to a week; IFA and ELISA are fast but require special reagents; RT-PCR, RRT-PCR, Gene chips detection technology is fast and sensitive , but all require special instruments and technical personnel, and the cost is relatively high, so it is not suitable for promotion and use at the grassroots level

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
  • RT-LAMP (reverse transcription loop-mediated isothermal amplification) visual detection kit for H1N1 subtype avian influenza viruses
  • RT-LAMP (reverse transcription loop-mediated isothermal amplification) visual detection kit for H1N1 subtype avian influenza viruses
  • RT-LAMP (reverse transcription loop-mediated isothermal amplification) visual detection kit for H1N1 subtype avian influenza viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, complete set of primers

[0047]The complete set of primers for loop-mediated isothermal amplification for detection or auxiliary detection of H1N1 subtype avian influenza virus in this embodiment is composed of the complete set of primers for detection or auxiliary detection of H1 subtype avian influenza virus and detection or auxiliary detection of N1 Primer sets for loop-mediated isothermal amplification of subtypes of avian influenza virus. Among them, the set of primers for loop-mediated isothermal amplification for detection or auxiliary detection of H1 subtype avian influenza virus consists of degenerate primer FIP-H1, degenerate primer BIP-H1, degenerate primer F3-H1, degenerate primer B3-H1 , degenerate primer LF-H1 and degenerate primer LB-H1, and their nucleotide sequences are respectively Sequence 1, Sequence 2, Sequence 3, Sequence 4, Sequence 5 and Sequence 6 in the sequence listing (Table 1). Among them, F3-H1 and B3-H1 are outer primers, F...

Embodiment 2

[0053] Example 2. Optimization of Reverse Transcription Loop-Mediated Isothermal Amplification Reagent for Detection or Assisted Detection of H1 Subtype and N1 Subtype Avian Influenza Virus

[0054] In this embodiment, strains A / Duck / Guangxi / 030D / 2009 (H1N1) and A / Duck / Guangxi / D / 2011 (H1N2) are used as samples to be tested for the following detection or auxiliary detection of H1 subtype and N1 subtype avian influenza The viral reverse transcription loop mediates the effect of isothermal amplification reagents:

[0055] Methods as below:

[0056] 1. RNA extraction of H1N1 subtype avian influenza virus

[0057] Total RNA was extracted from clinically isolated strain A / Duck / Guangxi / 030D / 2009 (H1N1) identified by hemagglutination test, hemagglutination inhibition test and sequence determination using TRIZOL method or RNA extraction kit.

[0058] 2. Optimization of RT-LAMP reaction conditions

[0059] Optimize the reagents that affect the efficiency of the amplification reaction...

Embodiment 3

[0066] Embodiment 3, utilize the RT-LAMP kit of detection or auxiliary detection H1 subtype, N1 subtype avian influenza virus to detect chicken source sample

[0067] A. In this example, the following RT-LAMP kits for detection or auxiliary detection of H1 subtype avian influenza virus were used for detection. The kit is a product obtained by assembling together RT-LAMP reaction reagent A, positive control A, negative control and fluorescent reagent for detection or auxiliary detection of H1 subtype avian influenza virus.

[0068] Among them, 19 μl of RT-LAMP reaction reagent for detection or auxiliary detection of H1 subtype avian influenza virus (hereinafter referred to as RT-LAMP reaction reagent A) is composed as follows: 2.5 μl 10-fold reaction buffer (20mM Tris-HCl, 10mM KCl, 10mM (NH 4 ) 2 SO 4 , 2mM MgSO 4 , 0.1%Triton X-100), 6mmol / L MgSO 4 , 1mmol / L betaine, outer primers F3-H1 and B3-H1 each 0.2μmol / L, inner primers FIP-H1 and BIP-H1 each 1.6μmol / L, loop primer...

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 an RT-LAMP (Reverse Transcription Loop-mediated isothermal amplification) visual detection kit for H1N1 subtype avian influenza viruses. The kit comprises an LAMP primer set for detection or auxiliary detection of H1 subtype and N1 subtype avian influenza viruses, wherein the primer set comprises degenerate primers FIP-H1, BIP-H1, F3-H1, B3-H1, LF-H1, LB-H1, FIP-N1, BIP-N1, F3-N1, B3-N1, LF-N1 and LB-N1, and the nucleotide sequences of the degenerate primers are respectively sequences 1 to 12 in a sequence table. The RT-LAMP visual detection kit has the advantages of good specificity, high sensitivity, simple instrument and equipment and easiness in operation, and convenience in result observation, and is particularly suitable for quick diagnosis and early screening of the H1 subtype and H1N1 subtype avian influenza viruses in grassroots veterinary stations, livestock farms and frontier ports.

Description

technical field [0001] The invention relates to an RT-LAMP visual detection kit for H1N1 subtype avian influenza virus. Background technique [0002] Avian influenza (Avian Influenza, AI) is the abbreviation of avian influenza, which is an infectious disease of poultry (poultry and wild birds) caused by the Orthomyxoviridae influenza virus type A avian influenza virus. According to the antigenicity of the surface glycoprotein (Hemagglutinin, HA) and neuraminidase (Neuraminidase, NA) of type A influenza virus, avian influenza virus (AIV) can be further divided into different subtypes, and 16 types of HA have been found so far. subtype and nine NA subtypes. AIV is an enveloped, single-stranded negative-strand, segmented RNA virus, and the viral genome consists of eight segmented negative-strand RNAs. The segmented nature of the AIV genome makes it possible that when two or more different subtypes of influenza viruses co-infect a cell, the nucleic acid fragments of the progen...

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
Inventor 谢芝勋彭宜刘加波谢志勤庞耀珊邓显文谢丽基范晴
Owner GUANGXI VETERINARY RES INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com