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Composition, kit and method for detecting and typing viruses causing respiratory tract infection and application of composition, kit and method

A composition and respiratory technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as difficult diagnosis, reduce psychological burden, block virus pandemics and outbreaks, and reduce waste Effect

Active Publication Date: 2020-04-28
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The pathogens of respiratory tract infection are extremely diverse. One clinical manifestation can be caused by multiple pathogens, and one pathogen can also cause multiple clinical manifestations, which brings great difficulties to early clinical diagnosis.

Method used

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  • Composition, kit and method for detecting and typing viruses causing respiratory tract infection and application of composition, kit and method
  • Composition, kit and method for detecting and typing viruses causing respiratory tract infection and application of composition, kit and method
  • Composition, kit and method for detecting and typing viruses causing respiratory tract infection and application of composition, kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1, primers and probes used in the present invention

[0073] The primers and probes used in the present invention are shown in Table 1 below

[0074]

[0075]

[0076] Among them, the fluorescent reporter group of the novel coronavirus 2019-nCoV probe is FAM; the fluorescent reporter group of influenza A virus is HEX; the fluorescent reporter group of influenza B virus is ROX, and the fluorescent reporter group of internal standard is CY5 , the 3' end of the probe also has a BHQ1 or BHQ2 quenching group.

Embodiment 2

[0077] Embodiment 2, detect the virus that causes respiratory tract infection and the method for typing

[0078] The detection samples of the present invention are throat swabs, sputum, alveolar lavage fluid, and blood. Virus nucleic acid is extracted by the magnetic bead method, and the following operations are performed in the sample processing room:

[0079] 2.1 Take an appropriate amount of 1.5 mL sterilized centrifuge tubes, mark the negative control, positive control and samples to be tested respectively, and add 300 μL RNA extraction solution 1 to each tube;

[0080] 2.2 Add 200 μL of the sample to be tested or the negative control and positive control to each tube; cover the tube cap, shake and mix for 10 seconds, and centrifuge briefly;

[0081] 2.3 Add 100 μL RNA Extraction Solution 2-mix to each tube (mix well and draw), shake and mix for 10 seconds, then let stand at room temperature for 10 minutes;

[0082] 2.4 Place the centrifuge tube on the separator after br...

Embodiment 3

[0098] Embodiment 3, the detection result of composition test positive control of the present invention

[0099] Using the composition in Table 1 of the present invention, according to the method described in Example 2, each target-positive plasmid is detected to simulate a clinical sample, and multiplex PCR detection is performed on a Hongshi fluorescent quantitative PCR instrument, and the results are as follows Figure 1~4 shown. Good amplification curves can be detected in each channel, indicating that the composition of the invention can detect and type viruses that cause respiratory tract infections.

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Abstract

The invention relates to the field of molecular biology detection, in particular to detection of novel coronavirus, influenza A virus and influenza B virus. The invention provides a composition for detecting the viruses, and meanwhile, the invention further provides a kit containing the composition, application of the composition and a method for detecting and typing the viruses causing respiratory tract infection. The composition is combined with a fluorescent probe method, three viruses causing respiratory tract infection can be detected and typed at the same time in one tube, and the composition, the kit and the method have the advantages of being low in cost, high in flux, short in consumed time, easy and convenient to operate and capable of avoiding false positive caused by cross among samples and environmental pollution.

Description

technical field [0001] The invention belongs to the field of molecular biology detection; specifically, it belongs to the field of detection of viruses causing respiratory tract infections; more specifically, the invention can simultaneously detect novel coronavirus 2019-nCoV, influenza A virus and influenza B virus, and its classification. Background technique [0002] Respiratory diseases caused by respiratory infections are common diseases in life, and the pathogens mainly include viruses, bacteria, mycoplasma, chlamydia, etc. Among them, common viral pathogens include influenza A virus and influenza B virus, which are also the most important pathogens of seasonal influenza; and another type of virus that causes respiratory tract infections is coronavirus. [0003] The envelope of influenza virus has two very important glycoproteins: hemagglutinin (HA) and neuraminidase (NA). Generally, there are 500 hemagglutinin spikes and 100 neuraminidases distributed on the surface ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2600/166C12Q2531/113C12Q2521/107C12Q2545/101C12Q2563/107C12Q1/6883
Inventor 戴立忠谭德勇邓中平孙青芝任小梅程星刘佳陈晓亮毛君杰
Owner SANSURE BIOTECH INC
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