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Composition, test kit and method for detecting and typing 10 kinds of respiratory tract related viruses, and application of composition

A respiratory tract and composition technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of low culture positive rate, harsh virus culture conditions, and difficulty in identifying and determining virus types, so as to improve detection efficiency. , to avoid false positives and environmental pollution, the effect of low cost

Pending Publication Date: 2020-10-23
SANSURE BIOTECH
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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, test kit and method for detecting and typing 10 kinds of respiratory tract related viruses, and application of composition
  • Composition, test kit and method for detecting and typing 10 kinds of respiratory tract related viruses, and application of composition
  • Composition, test kit and method for detecting and typing 10 kinds of respiratory tract related viruses, and application of composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Embodiment 1, primers and probes used in the present invention

[0076] The primers and probes used in the present invention are shown in Table 1 below:

[0077]

[0078]

[0079]

[0080] 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.

[0081] The fluorescent reporter group of the upstream primers of human parainfluenza virus type I, II, III and IV is FAM; the fluorescent reporter group of the upstream primers of influenza A virus is HEX; the fluorescent reporter group of the upstream primers 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

[0082] Embodiment 2, the method for detecting respiratory tract-associated virus and typing

[0083] 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:

[0084] 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;

[0085] 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;

[0086] 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;

[0087] 2.4 Place the centrifuge tube on the separator after brief centrifuga...

Embodiment 3

[0104] Embodiment 3, the detection result of composition test positive control of the present invention

[0105] 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 molecular biological detection, specifically to detection of novel coronavirus 2019-nCoV, influenza virus type A, influenza virus type B, human parainfluenza virus type I, human parainfluenza virus type II, human parainfluenza virus type III, human parainfluenza virus type IV, respiratory adenovirus, respiratory syncytial virus and human metapneumovirus. Meanwhile, the invention also provides a test kit containing the composition, an application of the composition and a method for detecting and typing respiratory tract related viruses. The composition disclosed by the invention combines a fluorescent probe method and a melting curve method, can be used for simultaneously detecting and typing the respiratory tract related viruses in one tube, is low incost, high in flux and less in time consumption, greatly improves the detection efficiency, is simple and convenient to operate, and avoids false positive result and environmental pollution caused bycross of samples through 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, human parainfluenza virus type I, human parainfluenza virus type II, human Detection and typing of parainfluenza virus type III, human parainfluenza virus type IV, respiratory adenovirus, respiratory syncytial virus and metapneumovirus. 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, fast transmission, short incubation period, and acute onset. They can cause a wide range of acute u...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2600/16C12Q2600/166C12Q2521/107C12Q2531/113C12Q2537/143C12Q2563/107C12Q2545/101
Inventor 邓中平谭德勇戴立忠陈晓亮程星任小梅胡娟
Owner SANSURE BIOTECH
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