Respiratory virus detection kit and detection method

A virus detection and respiratory technology, applied in the field of molecular biology, can solve the problems of long detection time, inability to cope with respiratory virus detection, and low sensitivity, and achieve the effects of sensitivity, specificity, strong practicability, and high detection throughput

Active Publication Date: 2011-09-14
深圳市赛尔生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, the traditional respiratory virus detection method has a long detecti

Method used

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  • Respiratory virus detection kit and detection method
  • Respiratory virus detection kit and detection method
  • Respiratory virus detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Design and prepare primers and probes

[0079] By analyzing the gene sequences of 8 common respiratory viruses, a relatively conservative sequence is selected as the target amplification fragment, and PCR amplification primers for amplifying the fragment and specifically binding to the fragment are designed according to the target amplification fragment the probe.

[0080] The 8 target amplification fragments and their corresponding PCR amplification primers and probes are as follows:

[0081] The first target amplified fragment is the nucleic acid fragment shown in SEQ ID NO.1 of the gene of SARS virus, the corresponding upstream primer sequence is shown in SEQ ID NO.9, and the downstream primer sequence is shown in SEQ ID NO.17 , the probe sequence is shown in SEQ ID NO.25.

[0082] The second target amplified fragment is the nucleic acid fragment shown in SEQ ID NO.2 of the gene of influenza A virus, the corresponding upstream primer sequence is shown in SEQ ID NO....

Embodiment 2

[0093] Respiratory virus detection kit and detection method

[0094] The detection kit includes the above-mentioned 8 sets of primers and 8 probes.

[0095] In a preferred embodiment, the detection kit also includes a positive quality control probe and a positive probe complementary to the positive quality control probe; the sequence of the positive quality control probe is shown in SEQ ID NO.33, and the positive probe's The sequence is shown in SEQ ID NO.34.

[0096] In this example, the 5' end of the positive probe was labeled with biotin.

[0097] Such as figure 1 The detection method of shown respiratory virus comprises the steps:

[0098] S10, extracting the nucleic acid in the sample to be tested

[0099] Nucleic acid was extracted using the column virus RNAOUT extraction kit (product number 71001-50) of Beijing Tianenze Gene Technology Co., Ltd.

[0100] The usage method is as follows:

[0101] 1. For liquid virus samples: Add 0.1-0.2 mL of liquid virus samples in...

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Abstract

The invention relates to a respiratory virus detection kit, and simultaneously provides a respiratory virus detection method using the kit. The kit contains 8 groups of primers and probes thereof, wherein in the primers, the 5' ends of the downstream primers are marked by using biotins. The detection method comprises the following steps of: amplifying nucleic acid in a sample to be detected by using the primers, and hybridizing the formed biotinylation products and the probes fixed on the surface of a chip to form a probe-biotinylation product complex; combining an alkaline phosphatase-streptavidin conjugate and the probe-biotinylation product complex, and performing color (bluish violet) generation reaction on nitroblue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate (NBT/BCIP) serving as a color development substrate and alkaline phosphatase; and detecting whether the sample to be detected contains respiratory viruses to be detected by judging whether the specific site develops color. The kit has high detection flux, is quick and sensitive and has strong specificity and very strong practicability.

Description

【Technical field】 [0001] The invention relates to the field of molecular biology, in particular to a respiratory virus detection kit and detection method. 【Background technique】 [0002] Common respiratory viruses include SARS (Severe Acute Respiratory Syndromes, Severe Acute Respiratory Syndrome) virus, A and B influenza viruses, parainfluenza virus I, II, III, respiratory fusion virus, adenovirus, etc., in many densely populated areas Public places such as railway stations, squares, etc. need to be tested for the above-mentioned viruses to prevent the rapid spread of the virus among the crowd and avoid large-scale outbreaks of respiratory diseases. [0003] However, traditional respiratory virus detection methods have a long detection time and low sensitivity, and cannot cope with sudden respiratory virus detection. 【Content of invention】 [0004] Based on this, it is necessary to provide a respiratory virus detection kit with shorter detection time and higher sensitivi...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 顾大勇黄彤文李永进张巍史蕾孙秋香刘春晓赵纯中杨燕秋徐云庆孙杰莫秋华杨泽庄卫东
Owner 深圳市赛尔生物技术有限公司
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