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Kit for isothermal detection of fungal pathogens of respiratory tract infection with enzyme-cleavage probe

A technology for pathogens and respiratory tracts, 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 time-consuming, high detection costs, laborious and other problems, and achieves the effect of simple operation.

Active Publication Date: 2022-08-05
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 isothermal detection of fungal pathogens of respiratory tract infection with enzyme-cleavage probe
  • Kit for isothermal detection of fungal pathogens of respiratory tract infection with enzyme-cleavage probe
  • Kit for isothermal detection of fungal pathogens of respiratory tract infection with enzyme-cleavage probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Determination of primer-probe combinations for multiplex nucleic acid detection of 8 fungal pathogens of respiratory tract infection

[0024] 1. Design of singleplex primers and probes

[0025] The present invention is effective against 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 the design software was used to design on the basis of the basic principles of primer-probe design, and the primer and probe combinations were screened for each pathogen target. The primer-probe combinations screened for each pathogen target are shown in Table 1.

[0026] Table 1. Primer-probe combinations screened for 8 fungal pathogens

[0027]

[0028]

[0029]

[0030] 2. Screening of single primers and probes

[0031] 2.1 The single reaction liquid system is shown in Table...

Embodiment 2

[0053] Embodiment 2. Preparation of the detection kit in the present invention and verification of the sensitivity and specificity of the kit

[0054] 1. Preparation of the Kit

[0055] Three nucleic acid reaction solutions, blank reference substances, and positive reference substances are co-packaged, and the product instruction manual is attached to obtain the kit of the present invention for the constant temperature detection of 8 high-incidence fungal pathogens in respiratory tract infections by the enzyme cleavage probe. 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 fluorophore labels of the probes for detecting each pathogen are shown in Table 7.

[0056] Table 6. Components of the kit

[0057]

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

[0059]

[0060] 2. The kit in the present invention detects 4 clinical samples ...

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Abstract

The invention provides a kit for the constant temperature detection of fungal pathogens of respiratory tract infection with an enzyme cleavage probe, which combines the primer probes of eight pathogen targets into three multiple nucleic acid reaction solutions. Using the nucleic acid detection kit provided in the present invention, three single-tube multiplex constant temperature amplification can be used to detect 8 kinds of fungal pathogens with high frequency in respiratory tract infections, and the kit is easy to operate, and bedside detection can be realized, and the entire detection time Within 30 minutes, it can quickly and accurately screen a variety of fungal pathogens in the sample to be tested.

Description

technical field [0001] The invention relates to the technical field of nucleic acid constant temperature detection, in particular to a kit for the constant temperature detection of respiratory tract infection fungal pathogens with an enzyme cleavage probe. Background technique [0002] Respiratory tract infection is a common disease with high incidence. Most of the fungal infections in respiratory tract infections are Candida, Aspergillus, Mucor, Pneumocystis jirovecii, etc. Among them, Candida albicans, Candida parapsilosis, Five species, Candida glabrata, Candida tropicalis, and Candida krusei, account for 95% of human fungal infections. In recent years, the use of broad-spectrum antibiotics has caused the incidence of fungal infections to increase year by year, resulting in a high mortality rate. Therefore, the realization of early and rapid specific diagnosis of fungal pathogens is very necessary to improve the prognosis of patients and improve the survival rate of patie...

Claims

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Application Information

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Patent Type & Authority Patents(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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