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Nucleic acid composition and kit for detecting drug resistance genes of mycobacterium tuberculosis and method for detecting drug resistance of mycobacterium tuberculosis

A technology of mycobacterium tuberculosis and nucleic acid composition, which is applied in the field of medical detection, can solve the problems of high cost, low throughput, complicated operation, etc., and achieve the effects of fast detection speed, high accuracy and good repeatability

Pending Publication Date: 2022-04-08
深圳汉亚生物科技合伙企业(有限合伙)
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Problems solved by technology

However, fluorescent quantitative PCR is limited by the type of fluorescence and reagent consumables, and the throughput is small and the cost is high; whole gene sequencing requires a professional technical foundation, and the operation is complicated, time-consuming and laborious; Can detect novel mutations, but with low sensitivity and poor reproducibility

Method used

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  • Nucleic acid composition and kit for detecting drug resistance genes of mycobacterium tuberculosis and method for detecting drug resistance of mycobacterium tuberculosis
  • Nucleic acid composition and kit for detecting drug resistance genes of mycobacterium tuberculosis and method for detecting drug resistance of mycobacterium tuberculosis
  • Nucleic acid composition and kit for detecting drug resistance genes of mycobacterium tuberculosis and method for detecting drug resistance of mycobacterium tuberculosis

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Embodiment 1

[0058] This embodiment provides a method for detecting drug resistance of Mycobacterium tuberculosis, the flow chart refers to figure 1 As shown, it includes the following steps:

[0059] 1. Extract the DNA of the sample to be tested.

[0060] Using a commercial Mycobacterium tuberculosis DNA extraction kit (Xiamen Zhishan Biotechnology Co., Ltd., Lab-Aid 824 Mycobacterium tuberculosis nucleic acid extraction Maxi reagent, Minxia Jibei No. 20170046), sample DNA was extracted according to the operating instructions. The extracted DNA needs to be frozen and stored, thawed before use, vortexed and mixed, but repeated freezing and thawing should not exceed 3 times. The samples to be extracted for DNA may be potentially infectious, and protective measures must be taken before operation.

[0061] 2. Primer design and amplification.

[0062] According to the main drug-resistant mutations in the resistance sites of three drugs, rifampicin, isoniazid, and ethambutol, published by WH...

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Abstract

The invention discloses a nucleic acid composition and kit for detecting drug resistance genes of mycobacterium tuberculosis and a method for detecting drug resistance of mycobacterium tuberculosis, and relates to the technical field of medical detection. According to the invention, a single base extension reaction technology is combined, to-be-detected mutation sites of katG315, rpoB435 and embB306 genes are determined, amplification primers and extension primers of the mutation sites are designed, after PCR amplification, digestion treatment and extension reaction, resin purification and nanoliter sample application are carried out to prepare a sample, mass spectrometry detection is carried out, and finally, site mutation details are determined through comparison. Therefore, the drug resistance of the mycobacterium tuberculosis to isoniazid, rifampicin and ethambutol is judged. The detection result can be widely applied to the fields of clinical examination, disease prevention, epidemic disease screening, import and export inspection and quarantine and the like.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a nucleic acid composition for detecting drug resistance genes of Mycobacterium tuberculosis, a kit and a method for detecting drug resistance of Mycobacterium tuberculosis. Background technique [0002] As of 2020, tuberculosis caused by Mycobacterium tuberculosis is still one of the major infectious diseases that endanger human life and health, and the multi-drug resistance of tuberculosis is the biggest difficulty and severe challenge in the treatment of modern tuberculosis. In the past 10 years, the resistance of Mycobacterium tuberculosis to clinical mainstream anti-tuberculosis drugs including isoniazid (INH), rifampicin (RIF) and ethambutol (EMB) has increased significantly. In the part of the drug resistance mechanism of Mycobacterium tuberculosis that has been elucidated, it is believed that rifampicin resistance is caused by the mutation of the determinant sit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6858C12N15/11
Inventor 宋希凡
Owner 深圳汉亚生物科技合伙企业(有限合伙)
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