Influenza A virus genotype detection kit

A type of influenza A virus and detection kit technology, applied in the biological field, can solve problems such as the absence of influenza A virus, and achieve the effects of preventing multiple operation contamination, clear results and strong specificity

Inactive Publication Date: 2018-07-03
SHANGHAI XINGYAO MED TECH DEV CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In China, there are influenza A virus nucleic acid detection kits (PCR-fluorescent probe method) that have obtained the national production license, but there is no fluorescent PCR detection kit for joint detection of influenza A virus subtypes on the market

Method used

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  • Influenza A virus genotype detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 A Genotyping Detection Kit for Influenza A Virus

[0042] 1. Extraction of Influenza Virus Nucleic Acid

[0043] Nucleic acid was extracted from human nasopharyngeal swab samples using the magnetic bead nucleic acid extraction reagent (nucleic acid extraction and purification reagent (body fluid RNA)) produced by Shanghai Fosun Changzheng Medical Science Co., Ltd.

[0044] 2. Reverse transcription and fluorescent RT-PCR amplification (50 μl system per person)

[0045] a. Preparation of one-step RT-PCR reaction solution:

[0046]

[0047] b. One-step RT-PCR reaction procedure:

[0048] [1] 50°C 25 min

[0049] [2] 95℃ for 5 minutes

[0050] [3] 95°C 15s

[0051] [4] 60°C 50s

[0052] [5] Go to [3] ,45 cycles

[0053] Collect fluorescence signals of FAM, HEX, ROX and CY5 channels in the fourth step

[0054] [6] End

[0055] 3. Detection:

[0056] The invention is applicable to ABI 7500 fluorescent quantitative PCR instrument for genotyping detection...

Embodiment 2

[0061] Example 2 clinical testing

[0062] Qualitative detection of 50 cases of clinical samples by fluorescent PCR was carried out with the above method, including 6 cases of seasonal influenza virus H1 subtype, with a positive detection rate of 12%; 7 cases of seasonal influenza virus H3 subtype, with a positive detection rate of 14%. %; 3 patients with H1N1 influenza virus (2009), the positive rate was 6%. The detection method and the kit of the present invention have strong specificity, high sensitivity, simple operation and high repeatability, and can detect seasonal influenza virus H1 subtype, H3 subtype and type A H1N1 influenza virus in human nasopharyngeal swab samples (2009) qualitative identification and detection. The present invention uses the magnetic bead method nucleic acid extraction reagent (nucleic acid extraction and purification reagent (body fluid RNA)) produced by Shanghai Fosun Changzheng Medical Science Co., Ltd. to extract nucleic acid from human nas...

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Abstract

The invention provides an influenza A virus genotype detection kit. The kit comprises RT-PCR reaction liquid, an RT-PCR enzyme mixture, an influenza A virus typing primer probe, an internal reference,a negative control, a critical positive control and a strongly positive control. The kit can directly carry out one-step RT-PCR on extracted influenza A virus RNA, can carry out fluorescent typing detection on influenza A viruses in a sample, takes an internal reference gene sequence as the internal reference and prevents pollution by utilizing UNG enzyme. The kit provided by the invention is simple in an amplification method, short in procedure and easy to operate, pollution is prevented, detection result specificity is strong, sensitivity is high, result is clear, reliability is high, and the kit provided by the invention can be applied to fluorescent type identification and detection on seasonal influenza virus H1 subtype and H3 subtype and influenza A virus subtype H1N1 (2009).

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a biological detection technology, in particular to a method for genotyping of influenza A virus (seasonal influenza virus H1 subtype, H3 subtype and influenza A H1N1 virus (2009)) detection Reagent test kit. Background technique [0002] Influenza virus, referred to as influenza virus, is a representative species of Orthomyxoviridae, including human influenza virus and animal influenza virus. Human influenza viruses can be divided into three types: Influenza A virus, Influenza B virus and Influenza C virus. Among them, the type A virus often undergoes antigenic variation, is highly contagious, spreads rapidly, and is extremely prone to widespread epidemics. According to the antigenicity of the outer membrane hemagglutinin (HA) and neuraminidase (NA) proteins of virus particles, influenza A virus can be divided into 16 H subtypes (H1-H16) and 9 N subtypes ( N1-N9), there hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/686C12Q1/701C12Q2521/107C12Q2561/101C12Q2545/107
Inventor 迟大利吴大治夏懿
Owner SHANGHAI XINGYAO MED TECH DEV CO LTD
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