Composition for detecting and typing pathogens causing respiratory tract infection, kit, method and application

A technology of respiratory tract and composition, applied in the direction of microorganism-based methods, biochemical equipment and methods, and resistance to vector-borne diseases, etc., can solve the problems of patients with respiratory tract infection and clinicians, difficult to identify pathogen types, and harsh pathogen culture conditions To avoid false positives and environmental pollution, reduce psychological burden, and reduce costs

Active 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 pathogens infected by clinical symptoms and routine laboratory tests, and the culture conditions of pathogens are relatively harsh, resulting in a low positive culture rate, and even some pathogens cannot be cultured under the current conditions. Great distress for patients and clinicians

Method used

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  • Composition for detecting and typing pathogens causing respiratory tract infection, kit, method and application
  • Composition for detecting and typing pathogens causing respiratory tract infection, kit, method and application
  • Composition for detecting and typing pathogens causing respiratory tract infection, kit, method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1, primers and probes used in the present invention

[0078] The primers and probes used in the present invention are shown in Table 1 below:

[0079]

[0080]

[0081]

[0082] Among them, the fluorescent reporter group of the influenza A virus probe and the respiratory adenovirus probe is FAM; the fluorescent reporter group of the new coronavirus 2019-nCoV probe and the influenza B virus probe is HEX; the respiratory syncytial virus and The fluorescent reporting group of the Mycoplasma pneumoniae probe is CY5, the fluorescent reporting group of the internal standard is ROX, and the 3' end of the probe also has a BHQ1 or BHQ2 quenching group.

Embodiment 2

[0083] Embodiment 2, detection causes the pathogen of respiratory tract infection and the method for typing

[0084] 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:

[0085] 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;

[0086] 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;

[0087] 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;

[0088] 2.4 Place the centrifuge tube on the separator afte...

Embodiment 3

[0103] Embodiment 3, the detection result of composition test clinical sample of the present invention

[0104] Using the composition in Table 1 of the present invention, according to the method described in Example 2, 411 samples were tested, including 1 part of the 2019 novel coronavirus cell culture solution, 25 positive cases of influenza A virus, and 25 cases of influenza B virus. 68 cases were positive, 77 cases were positive for respiratory adenovirus, 18 cases were positive for respiratory syncytial virus, and 27 cases were positive for mycoplasma pneumoniae. The overall results were consistent with the identification results of the Center for Disease Control and Prevention. In addition, the composition of the present invention was compared with the control method (the method using a commercially available kit as a control kit), and while the control method had false negatives or false positives, the detection effect of the composition of the present invention was even ...

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Abstract

The invention relates to the field of molecular biology detection, and particularly relates to detection of novel coronavirus 2019- nCoV, influenza A virus, influenza B virus, respiratory adenovirus,respiratory syncytial virus and mycoplasma pneumoniae. The invention provides a composition for detecting the pathogens. Meanwhile, the invention further provides a kit containing the composition, application of the composition and a method for detecting and typing the pathogens causing respiratory tract infection. The composition is combined with a fluorescent probe method, six pathogens causingrespiratory tract infection can be detected and typed at the same time in two tubes, and the advantages that the cost is low, the flux is high, the influence of interference substances is avoided, operation is easy and convenient, and false positive and environmental pollution caused by sample crossing are avoided are achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology detection; specifically, it belongs to the field of detection of pathogens causing respiratory infections; more specifically, the invention can simultaneously detect novel coronavirus 2019-nCoV, influenza A virus, influenza B virus, respiratory gland Viruses, respiratory syncytial virus and mycoplasma pneumoniae, and their typing. Background technique [0002] Most respiratory infectious diseases are clinically common and frequently-occurring diseases. Such diseases have similar clinical symptoms and epidemic characteristics, and it is difficult to identify and determine the type of pathogen infected by clinical symptoms and routine laboratory tests. Respiratory pathogens 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/689C12Q1/6851C12Q1/06C12R1/93C12R1/35
CPCC12Q1/701C12Q1/689C12Q1/6851C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107C12Q2600/112
Inventor 陈晓亮任小梅向江艳万旺廖石榴邓中平戴立忠
Owner SANSURE BIOTECH INC
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