Composition for detecting and typing respiratory tract related viruses, kit, use and method

A respiratory and composition technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of harsh virus culture conditions, difficult to identify and determine virus species, low positive rate of culture, etc., to achieve Avoid false positives and environmental pollution, low cost, and improve detection efficiency

Inactive Publication Date: 2020-06-23
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult to identify and determine the type of virus infected through clinical symptoms and routine laboratory tests, and the culture conditions of the virus are relatively harsh, resulting in a low positive culture rate, and even some viruses cannot be cultured under the current conditions. Great distress for patients and clinicians

Method used

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  • Composition for detecting and typing respiratory tract related viruses, kit, use and method
  • Composition for detecting and typing respiratory tract related viruses, kit, use and method
  • Composition for detecting and typing respiratory tract related viruses, kit, use and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1, primers and probes used in the present invention

[0067] The primers and probes used in the present invention are shown in Table 1 below:

[0068]

[0069]

[0070]

[0071] Among them, the fluorescent reporter group of the novel coronavirus 2019-nCoV probe is FAM; the fluorescent reporter group of the influenza B virus probe is HEX; the fluorescent reporter group of the respiratory adenovirus probe is ROX, and the 3' of the probe is The end also has a BHQ1 or BHQ2 quenching group.

[0072] The fluorescent reporter group of the upstream primer of influenza A virus is FAM; the fluorescent reporter group of the upstream primer of parainfluenza virus is HEX; the fluorescent reporter group of the upstream primer of respiratory syncytial virus is ROX; the fluorescent reporter group of metapneumovirus is CY5. The fluorescent reporter groups of the fluorescent primer groups are all labeled on a T base in the primers.

Embodiment 2

[0073] Embodiment 2, the method for detecting respiratory tract-associated virus and typing

[0074] 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:

[0075] 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;

[0076] 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;

[0077] 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;

[0078] 2.4 Place the centrifuge tube on the separator after brief centrifu...

Embodiment 3

[0095] Embodiment 3, the detection result of composition test positive control of the present invention

[0096] 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. It can be seen from the figure that both the fluorescent probe method and the melting curve method can detect the corresponding target, which proves that the composition of the present invention can detect and type the respiratory tract-associated virus.

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Abstract

The invention relates to the field of detection of molecular biology and more specifically relates to detection on novel coronavirus 2019-nCoV, influenza A virus, influenza B virus, parainfluenza virus, respiratory adenovirus, respiratory syncytial virus and metapneumovirus. Meanwhile, the invention further provides a kit containing the composition, use of the composition and a method for detecting and typing respiratory tract related viruses. According to the composition provided by the invention, in combination with a fluorescent probe method and a melting curve method, the respiratory tractrelated viruses can be simultaneously detected and typed in one tube, the cost is low, the flux is high, the consumed time is short, and the efficiency of detection is greatly increased; and the operation is simple and convenient, and false positive and environmental pollution caused by crossing among samples is avoided due to single-tube operation.

Description

technical field [0001] The invention belongs to the field of molecular biology detection, and more specifically, relates to novel coronavirus 2019-nCoV, influenza A virus, influenza B virus, parainfluenza virus, respiratory adenovirus, respiratory syncytial virus and metapneumonia Virus detection and typing. Background technique [0002] Most respiratory infectious diseases are clinically common and frequently-occurring diseases. There are many types of pathogens that cause respiratory diseases, and it is difficult to identify the type of pathogens infected by clinical symptoms and routine laboratory tests. Respiratory viruses can be transmitted through the air. The pathogens that cause acute respiratory diseases have the characteristics of strong infectivity, rapid transmission, short incubation period, and acute onset. They can cause a wide range of acute upper and lower respiratory diseases and seriously endanger human health. Common respiratory viruses include: [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/701C12Q2600/166C12Q2531/113C12Q2563/107C12Q2521/107C12Q2545/101Y02A50/30
Inventor 谭德勇邓中平戴立忠任小梅陈晓亮刘佳程星孙青芝
Owner SANSURE BIOTECH INC
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