Kit for nucleic acid combined detection of influenza virus A, influenza virus B and respiratory syncytial virus

A technology of influenza B virus and influenza A virus, which is applied in the field of joint detection kits for influenza A, influenza B virus and respiratory syncytial virus nucleic acid, can solve the problem of losing the ability to be re-amplified, and there is no respiratory syncytial virus Combined detection of fluorescent PCR detection kits, false positives and other issues, to achieve the effects of short amplification time, prevention of multiple operation contamination, and simple steps

Inactive Publication Date: 2016-03-16
SHANGHAI XINGYAO MED TECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Four domestic companies have developed joint detection reagents for influenza A virus and influenza B virus, all of which have been licensed by the state, but there is no fluorescent PCR detection kit for joint detection of respiratory syncytial virus
[0011] At the same time, in order to avoid false positives in the reaction results, using the anti-pollution characteristics of UNG enzyme, dUTP is used instead of dTTP in the PCR amplification react

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 A combined detection kit for influenza A, B and respiratory syncytial virus nucleic acid

[0043] 1. Extraction of Respiratory Virus Nucleic Acid

[0044] Nucleic acid was extracted from human nasopharyngeal swab samples using the magnetic bead purification reagent produced by Shanghai Fosun Changzheng Medical Science Co., Ltd.

[0045] 2. Reverse transcription and fluorescent RT-PCR amplification (50 μl system per person)

[0046] [0046] RT-PCR reaction solution [0047] 25 μl [0048] RT-PCR Enzyme Mix [0049] 1 μl [0050] Respiratory virus primer probe [0051] 4 μl [0052] template [0053] 20 μl [0054] Reaction volume: [0055] 50 μl

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

[0048] [1]50℃25min

[0049] [2]95℃5min

[0050] [3]95℃15s

[0051] [4]60℃50s

[0052] [5]Goto[4], 45cycles

[0053] In the fifth step, the fluorescence signals of FAM, HEX, CY3 and CY5 channels are collected.

[00...

Embodiment 2

[0061] Example 2 clinical testing

[0062] Using the above method, 50 cases of clinical samples were qualitatively detected by fluorescent PCR, including 15 cases of influenza A virus patients, with a positive detection rate of 30%; 4 cases of influenza B virus patients, with a positive detection rate of 8%; respiratory syncytial virus There were 12 patients, and the positive rate was 24%. The detection method and kit of the present invention have strong specificity, high sensitivity, simple operation and high repeatability, and can qualitatively identify influenza A virus, influenza B virus and respiratory syncytial virus in human nasopharyngeal swab samples , Detection. The present invention uses the magnetic bead method purification reagent produced by Shanghai Fosun Changzheng Medical Science Co., Ltd. to extract nucleic acid from human nasopharyngeal swab samples, thereby ensuring the purity of the template. At the same time, using one-step fluorescent RT-PCR technolo...

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PUM

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Abstract

The invention provides a kit for nucleic acid combined detection of the influenza virus A, the influenza virus B and the respiratory syncytial virus. The kit comprises RT-PCR reaction liquid, an RT-PCR enzyme mixture, a respiratory virus primer probe, internal reference, negative contrast, clinical positive contrast and strong positive contrast. The one-step RT-PCR reaction can be directly conducted on the well-extracted respiratory virus nucleic acid, the influenza virus A, the influenza virus B and the respiratory syncytial virus in a sample can be detected in a classified and qualitative mode, the gene of the internal reference serves as the internal contrast, and contamination is prevented through UNG enzymes. The kit is simple in one-step amplification method, short in procedure, easy and convenient to operate, capable of preventing contamination, high in detection result specificity, high in sensitivity, clear in result, high in credibility, and capable of being used for qualitatively authenticating and detecting the influenza virus A, the influenza virus B and the respiratory syncytial virus in a human nose pharynx-mop sample.

Description

Technical field [0001] The present invention is a biotechnology field, involving a biological testing technology, especially involving a combination of type A, B virus and respiratory tract hypotenocyte virus nucleic acid. Background technique [0002] VirusesAssociateDWithRespiratoryinfections refers to the general term that mainly uses the respiratory tract as an invasion of the portal.90-95%of clinical acute respiratory infections are caused by this group of poison.The respiratory virus is mostly a single -chain RNA (SSRNA) virus. It generally has envelope and thorns. It is mainly transmitted through the respiratory tract. It is positioned as the respiratory tract. It can be infected repeatedly and high population.Among them, Influenzavirus and respiratory tract sympathetic virus (RSV) are clinically and important respiratory viruses. [0003] The form of respiratory virus virus is spherical, the genome is a linear, regardless of the section of the single -chain RNA (SSRNA),...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701C12Q1/686
Inventor 迟大利吴大治夏懿
Owner SHANGHAI XINGYAO MED TECH DEV CO LTD
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