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Respiratory pathogen multi-detection reagent kit

A multiple detection and respiratory technology, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of cumbersome operation and long operation time

Inactive Publication Date: 2019-02-19
上海捷诺生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Single-plex PCR is the earliest established PCR method. It only detects a single DNA template, so it can only detect one pathogen. However, the products using multiple PCR methods on the market can only detect a few pathogens at the same time at the same time. Long and cumbersome to operate

Method used

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  • Respiratory pathogen multi-detection reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0155] Embodiment 1 A kind of kit for detecting sixteen kinds of respiratory pathogens

[0156] Sixteen respiratory pathogens including 12 RNA viruses (influenza A virus, influenza B virus, influenza A H1N1 virus, respiratory syncytial virus types A and B, parainfluenza virus-1 / -2 / -3, coronavirus OC43, coronavirus 229E, rhinovirus and human metapneumovirus), 2 DNA viruses (adenovirus and bocavirus), and 2 bacteria (Mycoplasma pneumoniae and Bordetella pertussis).

[0157] The kit includes

[0158] (1) Amplification primer set, including the primers shown in the table below, used to pre-amplify the corresponding pathogen target DNA fragment

[0159]

[0160]

[0161] (2) Specific primer set, including the primers shown in the table below, used to make the target DNA fragment corresponding to the pathogen carry specific probe detection sequence and internal fluorescent label

[0162]

[0163]

[0164]

[0165]

[0166] (3) Detection probe set, including the d...

Embodiment 2

[0171] Embodiment 2 A method for detecting sixteen kinds of respiratory pathogens

[0172] The above sixteen respiratory pathogens include 12 RNA viruses (influenza A virus, influenza B virus, influenza A H1N1 virus, respiratory syncytial virus A and B, parainfluenza virus-1 / -2 / -3 , coronavirus OC43, coronavirus 229E, rhinovirus and human metapneumovirus), 2 DNA viruses (adenovirus and bocavirus) and 2 bacteria (Mycoplasma pneumoniae and Bordetella pertussis).

[0173] The detection kit used is the kit described in Example 1 of the present invention.

[0174] Samples to be tested: 9 tissue samples from different sources that are known to be infected with respiratory pathogens (within the above-mentioned 16 respiratory pathogens), and the sample numbers are 1-9.

[0175] Detection method:

[0176] (1) Sample processing: add 5 μl of internal quality control substance for every 200 μl sample, and then extract nucleic acid;

[0177] (2) Negative control: 200 μl preservation sol...

Embodiment 3

[0202] Embodiment 3 A method for detecting sixteen kinds of respiratory pathogens

[0203] The detection kit used is the kit described in Example 1 of the present invention;

[0204] Samples to be tested: Sixteen kinds of pathogenic whole viruses or whole bacterial nucleic acids respectively stored in the sample preservation solution, one of which was taken, and 5 samples were prepared by gradient dilution;

[0205] The above-mentioned 5 samples were detected according to the detection method in Example 2, and 16 kinds of pathogens were detected sequentially. According to the detection limit of the corresponding pathogen when the positive rate was greater than or equal to 95%, by using the QX200TMDroplet Digital of Bio-Rad Company The absolute copy number obtained by TMPCR is used to calculate the detection limit), and the following table shows the detection limits of the sixteen kinds of pathogens tested:

[0206] Pathogen

[0207] It can be seen from the detection...

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Abstract

The invention discloses a respiratory pathogen multi-detection reagent kit. The respiratory pathogen multi-detection reagent kit has the advantages that the respiratory pathogen multi-detection reagent kit is based on multi-PCR (polymerase chain reaction) technologies, detection results can be determined by the aid of fluorescence resonance energy transfer via the melting temperature ranges, the respiratory pathogen multi-detection reagent kit can be used for qualitatively simultaneously detecting 16 types of respiratory pathogens, the 16 types of respiratory pathogens include 12 types of RNA(ribonucleic acid) viruses (influenza A viruses, influenza B viruses, H1N1 influenza A viruses, type A and type B respiratory syncytial viruses, type -1 / -2 / -3 parainfluenza viruses, type OC43 coronaviruses, type 229E coronaviruses, rhinoviruses and human metapneumovirus), 2 types of DNA (deoxyribonucleic acid) viruses (adenoviruses and bocavirus) and 2 types of bacteria (mycoplasma pneumoniae andbordetella pertussis), the respiratory pathogen multi-detection reagent kit is high in detection sensitivity, and the sensitivity even can reach 1 copy / reaction; the multi-detection reagent kit is good in specificity, and negative results of pathogens which have identical sampling sites and similar pathogenic mechanisms and are not in the detection range of the respiratory pathogen multi-detectionreagent kit can be obtained; the respiratory pathogen multi-detection reagent kit is short in operation time and easy to operate and can be used for quickly detecting the 16 types of respiratory pathogens in a single tube of a reaction system, the results are clear and are easy to interpret, and the like.

Description

technical field [0001] The invention relates to a detection kit for respiratory pathogens, in particular to a multiple detection kit for respiratory pathogens. Background technique [0002] Acute respiratory infection is the most prevalent infectious disease in adults and children. Respiratory tract infection is divided into upper respiratory tract infection and lower respiratory tract infection. Upper respiratory tract infection includes runny nose, conjunctivitis, pharyngitis, sinusitis and other symptoms. Both viruses and bacteria can cause acute upper respiratory tract infections, which pose a great challenge for diagnosis due to the wide variety of pathogens. At present, the main methods for the detection of respiratory pathogens include: pathogen isolation and culture methods, immunological methods and molecular detection methods. [0003] Pathogen isolation and culture is the gold standard for the detection of respiratory pathogens. It has high specificity, but low...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12Q1/04C12N15/11C12R1/35C12R1/01
Inventor 夏小凯程鲁向王瑜新秦佳华董江锴古斯·西蒙
Owner 上海捷诺生物科技股份有限公司
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