Method for detecting various respiratory viruses based on gene membrane chip

A technology for virus detection and respiratory tract, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc. It can solve the problems of slow detection speed, low sensitivity and high cost of respiratory virus, and achieve the prevention of aerosol Pollution, high detection sensitivity, and simple operation

Pending Publication Date: 2021-10-19
SICHUAN HUAHAN TRIO BIOTECH CO LTD
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Problems solved by technology

[0008] The main purpose of the present invention is to provide a method for the detection of multiple respiratory viruses based on gene membrane chips, which solves the probl

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  • Method for detecting various respiratory viruses based on gene membrane chip
  • Method for detecting various respiratory viruses based on gene membrane chip
  • Method for detecting various respiratory viruses based on gene membrane chip

Examples

Experimental program
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experiment example

[0115] Experimental example: According to the established detection system, specificity test, detection limit and stability test are carried out to test the detection performance of the system. The probe layout is as follows: image 3 shown;

[0116] Specificity test: Use the adjusted multiplex PCR primer Mix to amplify the nucleic acids of influenza A, influenza B, syncytial virus, adenovirus, rhinovirus, parainfluenza virus, Mycoplasma pneumoniae, Rickettsia, and Chlamydia pneumoniae respectively; hybridization test The result is as Figure 4 As shown, the percentage of samples that can be rapidly analyzed to obtain a negative test in the total number of healthy people;

[0117] Detection limit: using IA-1.0E+03copies / mL, IB-1.0E+04copies / mL, RSV-1.0E+04copies / mL, HAdV-1.0E+04copies / mL, HRV-1.0E+04copies / mL, HPIV-1.0E+04copies / mL critical concentration of viral nucleic acid for multiple PCR loading, amplification and hybridization detection; the results are as follows Fi...

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Abstract

The invention discloses a method for detecting various respiratory viruses based on a gene membrane chip. The method comprises the following detection steps: pretreating a sample: taking a nasal/pharyngeal secretion swab into a preservation tube containing a sample preservation solution, fully washing, sucking all the solution into a new centrifuge tube, centrifuging, and discarding the supernatant; extracting nucleic acid: adding the extracting solution and protease K into a centrifugal tube, heating and centrifuging after vortex oscillation, and taking supernate for PCR; carrying out multiple PCR sample adding and amplification: taking multiple respiratory viruses PCR Mix, a nucleic acid extract solution and enzyme-free water, and carrying out multiple PCR sample adding and amplification; performing membrane chip hybridization: performing hybridization reaction on the multiplex PCR amplification product and a membrane chip through a full-automatic membrane chip nucleic acid molecule hybridization instrument; and issuing a detection result report: directly issuing the detection result report through an automatic analysis system of the full-automatic membrane chip nucleic acid molecule hybridization instrument. According to the invention, multiple PCR and reverse dot blot hybridization technologies are combined, so that multiple respiratory pathogens in a single sample can be simultaneously detected through a single reaction, and a detection result report can be automatically issued.

Description

technical field [0001] The invention relates to a method for detecting multiple respiratory viruses based on a gene membrane chip, and belongs to the technical field of virus detection. Background technique [0002] Respiratory tract infection is one of the common clinical diseases. The main pathogens include bacteria, viruses, mycoplasma, chlamydia, fungi, etc., and more than 90% of them are caused by respiratory virus infection. , the elderly and those with low immune function, the common ones are influenza A virus, influenza B virus, respiratory syncytial virus, respiratory adenovirus, rhinovirus, parainfluenza virus, etc. Most of them are highly infectious and spread quickly. The incubation period is short, the onset is acute, and the clinical manifestations are similar. All patients present with fever and symptoms of upper respiratory tract infection. It is difficult to distinguish the infectious pathogen from the clinical manifestations alone. [0003] At present, the...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6837C12N15/11
CPCC12Q1/701C12Q1/6837C12Q2600/166C12Q2600/16C12Q2531/113C12Q2537/143C12Q2521/107C12Q2521/531C12Q2565/501C12Q2545/101
Inventor 周航李文静郎小亮黄盈黄广平
Owner SICHUAN HUAHAN TRIO BIOTECH CO LTD
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