Kit for constant-temperature detection of respiratory tract infection bacterial pathogens by enzyme digestion probes

A pathogen and kit technology, which is applied in the field of kits for the constant temperature detection of bacterial pathogens of respiratory tract infections with enzyme-cleavage probes, can solve the problems of difficult to achieve sensitivity and specificity, etc.

Active Publication Date: 2021-05-11
GENERAL HOSPITAL OF PLA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, most of the current nucleic acid constant temperature amplification methods are difficult to perform single-tube multiple detection, or the sensitivity and specificity of multiple detection are difficult to achieve the effect of single multiple detection.

Method used

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  • Kit for constant-temperature detection of respiratory tract infection bacterial pathogens by enzyme digestion probes
  • Kit for constant-temperature detection of respiratory tract infection bacterial pathogens by enzyme digestion probes
  • Kit for constant-temperature detection of respiratory tract infection bacterial pathogens by enzyme digestion probes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. Determination of primer probe combinations for multiple detection of nucleic acid of 12 kinds of bacterial pathogens in respiratory tract infection

[0028] 1. Design of single primer and probe

[0029] The present invention is effective against Staphylococcus aureus (SA), Pseudomonas aeruginosa (PA), Acinetobacter baumannii (Aba), Klebsiella pneumoniae (Kpn), Haemophilus influenzae (HI), Legionella pneumophila (LP), Enterococcus (Ec), Streptococcus pneumoniae (SP), Stenotrophomonas maltophilia (Sme), Burkholderia cepacia (BC), Escherichia coli (Ecoli), Stellaris Analyzing the gene sequence of bacterial pathogens such as NuA, using the design software to design based on the basic principles of primer and probe design, and screening the combination of primers and probes for each pathogen target, and each pathogen target was screened The three sets of primer-probe combinations are shown in Table 1.

[0030] Table 1: Sequence information of 12 kinds of bacteri...

Embodiment 2

[0059] Example 2, the preparation of the detection kit in the present invention and the verification of the sensitivity and specificity of the kit

[0060] 1. Preparation of kits

[0061] The 4 nucleic acid reaction solutions, the blank control substance, and the positive control substance are packaged together, and the product instruction manual is attached to obtain a kit for the constant temperature detection of 12 high-occurrence bacterial pathogens in respiratory tract infections by the enzyme-cutting probe in the present invention. The compositions of the nucleic acid reaction solution, the blank control substance and the positive control substance are shown in Table 6 below, and the fluorescent group labels of the probes for detecting various pathogens are shown in Table 7.

[0062] Table 6. Kit composition list

[0063]

[0064] Table 7. Instructions for the labeling of each probe fluorophore in the kit

[0065]

[0066]

[0067] 2. The kit in the present in...

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Abstract

The invention provides a kit for constant-temperature detection of respiratory tract infection bacterial pathogens by enzyme digestion probes. Primer probes of 12 pathogen targets are combined into four triple nucleic acid reaction solutions, and the probe for detecting each bacterial pathogen is a probe containing an RNA base and comprises 6 constant-temperature amplification primers. By utilizing the nucleic acid detection kit, detection of 12 bacterial pathogens with high frequency in respiratory tract infection can be achieved through four-single-tube multi-isothermal amplification, the kit is simple and convenient to operate, bedside detection can be achieved, the whole detection time is within 30 minutes, and screening of various bacterial pathogens in a sample to be detected can be quickly and accurately achieved.

Description

technical field [0001] The invention relates to the technical field of constant temperature detection of nucleic acids, in particular to a kit for constant temperature detection of bacterial pathogens of respiratory tract infection by enzyme-cutting probes. Background technique [0002] Respiratory tract infection is a common and high incidence clinically. Bacterial infection is a common cause of respiratory tract disease. There are many pathogenic bacteria that cause respiratory tract bacterial infection. Among adult respiratory tract bacterial infections, Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae, Haemophilus influenzae, Legionella pneumophila, Enterococcus, Streptococcus pneumoniae, Stenophagous maltophilia, Burkholderia cepacia, Escherichia coli, These 12 species of Nocardia astrocytes are the most common. Therefore, the realization of specific and rapid screening and detection of these 12 common bacterial pathogens in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12Q1/10C12Q1/14C12N15/11C12R1/445C12R1/19C12R1/01C12R1/385C12R1/46C12R1/38C12R1/21C12R1/22
CPCC12Q1/689C12Q1/6844C12Q2600/16C12Q2537/143
Inventor 解立新王凯飞刘利成王华贵蔡金玉唐奇陶毅冯华华王鹏志
Owner GENERAL HOSPITAL OF PLA
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