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Multiple RT-RPA primer combination for influenza A virus detecting and H1 and H3 typing and application thereof

A kind of influenza A virus, RT-RPA technology, applied in the field of molecular biology detection

Active Publication Date: 2018-06-22
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At present, RPA technology has been used for the rapid detection of various pathogenic organisms; however, the application of multiple RPA technology to the detection and typing of influenza A virus has not been reported.

Method used

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  • Multiple RT-RPA primer combination for influenza A virus detecting and H1 and H3 typing and application thereof
  • Multiple RT-RPA primer combination for influenza A virus detecting and H1 and H3 typing and application thereof
  • Multiple RT-RPA primer combination for influenza A virus detecting and H1 and H3 typing and application thereof

Examples

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Embodiment Construction

[0071] The technical solution of the present invention will be described in further detail below through specific examples, but it must be pointed out that the following examples are only used to describe the content of the invention, and do not constitute limitations to the protection scope of the present invention.

[0072] All solvents and reagents used in this example are commercially available products, primers and probes were synthesized by Shanghai Sangon Biotechnology Co., Ltd. (Shanghai); viral RNA extraction kit (Code No. DP315-R) was purchased from Tiangen Biochemical Technology Co., Ltd. (Beijing); TwistAmp Basic Kit was purchased from TwistDx Company (UK); reverse transcriptase AMVReverseTranscriptase (Code No.M5108) and RNase inhibitor (Code No.N2111) were purchased from Promega Company (USA, WI, Madison) .

[0073] 1. Preparation of Standards

[0074] (1) Nucleic acid extraction

[0075]Referring to the instructions of the viral RNA extraction kit (Tiangen Bio...

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Abstract

The invention belongs to the field of molecular biological detection, and relates to a multiple RT-RPA primer combination for influenza A virus detecting and H1 and H3 typing and application thereof.The multiple RT-RPA primer combination for influenza A virus detecting and H1 and H3 typing comprises an RT-RPA primer for detecting an influenza A virus Matrix gene, an H1 subtype HA gene and an H3 subtype HA gene, and the sequences are sequentially shown in SEQ ID NO.1-NO.6. The multiple RT-RPA primer combination can be used for detecting an influenza A virus and verifying H1 and H3 subtypes. According to the multiple RT-RPA primer combination, a multiple RT-RPA method is adopted for detecting the influenza A virus and verifying the H1 and H3 subtypes for the first time, and the combinationis short in consumed time, high in sensitivity and specificity and capable of being quickly and effectively used for influenza A virus detecting and H1 and H3 typing.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, relates to detection of influenza A virus and its subtype analysis method, in particular to a multiple RT-RPA (reverse transcription recombinase) for detection of influenza A virus and typing of H1 and H3 Polymerase amplification technology) primer combination and its application. Background technique [0002] Influenza virus (Influenza virus) is a common pathogenic organism of respiratory infectious diseases. It has extremely high morbidity and mortality. human health and life. According to incomplete statistics, the global annual seasonal influenza virus infection rate is 5-10% for adults and 20-30% for infants and young children, causing 250,000 to 500,000 deaths. In taxonomy, influenza viruses are mainly divided into three categories: A, B and C, all of which belong to Orthomyxoviridae, and are 8-segment, single-stranded negative-sense RNA viruses. A virus) has strong v...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2600/16C12Q2531/119C12Q2537/143C12Q2521/507C12Q2522/101C12Q2521/101
Inventor 李晓军孙宁王卫萍姚新月陈芳芳杨阳王洁
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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