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Kit for detecting fungal pathogens of respiratory tract infection at constant temperature by using enzyme digestion probes

A pathogen and kit technology, which is applied in the field of kits for the constant temperature detection of fungal pathogens of respiratory tract infections with enzyme cleavage probes, can solve the problems of laborious, time-consuming and high detection costs, achieve simple operation, rapid and accurate screening, and realize bedside detection. Effect

Active Publication Date: 2021-04-09
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 can only perform single-tube single-plex detection, which will be time-consuming, laborious, and costly for the screening and detection of multiple targets.

Method used

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  • Kit for detecting fungal pathogens of respiratory tract infection at constant temperature by using enzyme digestion probes
  • Kit for detecting fungal pathogens of respiratory tract infection at constant temperature by using enzyme digestion probes
  • Kit for detecting fungal pathogens of respiratory tract infection at constant temperature by using enzyme digestion probes

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1. Determination of the combination of primers and probes for the multiple detection of nucleic acids of 8 kinds of fungal pathogens in respiratory tract infection

[0024] 1. Design of single primer and probe

[0025] The present invention is to Candida albicans (CA), Candida parapsilosis (Cpa), Candida glabrata (Cg), Candida tropicalis (Ctr), Candida krusei (Kr), Aspergillus (Asp), Mucor ( Muc), Pneumocystis jirovecii (PJ) and other fungal pathogen gene sequences were analyzed, and based on the basic principles of primer and probe design, the design software was used to design, and the combination of primers and probes was screened for each pathogen target. The primer-probe combinations screened for each pathogen target are shown in Table 1.

[0026] Table 1.8 Primer probe combinations screened for fungal pathogens

[0027]

[0028]

[0029]

[0030] 2. Screening of single primers and probes

[0031] 2.1 The single reaction solution system is shown...

Embodiment 2

[0053] Example 2, the preparation of the detection kit in the present invention and the verification of the sensitivity and specificity of the kit

[0054] 1. Preparation of kits

[0055] The 3 nucleic acid reaction solutions, the blank control substance, and the positive control substance are packaged together, and the product instructions are attached to obtain a kit for the constant temperature detection of 8 kinds of high-occurrence fungal 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.

[0056] Table 6. Kit composition list

[0057]

[0058] Table 7. Instructions for the labeling of each probe fluorophore in the kit

[0059]

[0060] 2. The kit in the present invention dete...

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Abstract

The invention provides a kit for detecting fungal pathogens of respiratory tract infection at constant temperature by using enzyme digestion probes. Primer probes of eight pathogen targets are combined into three multiple nucleic acid reaction solutions. By using the nucleic acid detection kit provided by the invention, eight fungal pathogens with high frequency in respiratory tract infection can be detected through multiple constant-temperature amplification of three single tubes; the kit is simple and convenient to operate, bedside detection can be realized, the whole detection time is within 30 minutes, and screening of multiple fungal pathogens in a sample to be detected can be quickly and accurately realized.

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 fungal pathogens of respiratory tract infection by enzyme-cutting probes. Background technique [0002] Respiratory tract infection is a common high-incidence disease. In respiratory tract infection, fungal infections are mostly Candida, Aspergillus, Mucor, Pneumocystis jirovecii and other species, among which Candida albicans, Candida parapsilosis, Candida glabrata, Candida tropicalis, and Candida krusei account for 95% of human fungal pathogenic infections. In recent years, the use of broad-spectrum antibiotics has caused the incidence of fungal infections to increase year by year, and the resulting mortality remains high. Therefore, early and rapid specific diagnosis of fungal pathogens is very necessary to improve the prognosis of patients and increase the survival rate of patients. [0003] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6844C12Q1/04C12R1/72C12R1/725C12R1/66C12R1/74C12R1/785C12R1/645
CPCC12Q1/6895C12Q1/6844C12Q2600/16C12Q2537/143C12Q2563/107C12Q2545/113
Inventor 王凯飞解立新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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