CRISPR/Cas分子诊断技术在结核分枝杆菌耐药性检测中的研究进展

doi:10.19982/j.issn.1000-6621.20240523

摘要/Abstract

摘要：

结核病是全球公共卫生面临的一项严峻挑战。我国是结核病高负担国家,也是耐多药和利福平耐药结核病高负担国家。发展快速、灵敏的结核分枝杆菌耐药性检测方法对结核病的控制和治疗具有重要意义。成簇的规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)作为细菌适应性免疫的组成部分,能够高效、特异性地识别和切割外源核酸靶标;同时某些CRISPR相关蛋白(Cas)具有反式切割活性,这为建立CRISPR/Cas结核病耐药性诊断技术开辟了新的研究领域。本文对CRISPR/Cas技术检测结核分枝杆菌及其耐药性的最新研究动态进行综述,并探讨该领域的未来发展趋势。

关键词: 分枝杆菌,结核, 结核,抗多种药物性, 诊断技术, 分子生物学检测, 综述

Abstract:

Tuberculosis (TB) remains a significant global public health challenge. China is a high-burden country of TB and also a country with high burden of multidrug-resistant and rifampin-resistant TB. The development of rapid and sensitive methods for detecting the drug resistance of Mycobacterium tuberculosis is of great significance for the control and treatment of TB. Clustered regularly interspaced short palindromic repeats (CRISPR) as a component bacterial adaptive immunity can efficiently and specifically recognize and cleave exogenous nucleic acid targets. Meanwhile, some CRISPR-associated proteins (Cas) have trans-cleavage activity, paving the way for novel applications in CRISPR/Cas-based diagnostics for TB drug resistance. Therefore, this review summarizes the latest advancements in CRISPR/Cas technologies for detection Mycobacterium tuberculosis and its drug resistance, and discusses the future trends in this field.

Key words: Mycobacterium tuberculosis, Tuberculosis, multidrug-resistant, Diagnose technology, Molecular diagnostic techniques, Review

中图分类号:

王苑柠, 杜宗敏. CRISPR/Cas分子诊断技术在结核分枝杆菌耐药性检测中的研究进展[J]. 中国防痨杂志, 2025, 47(5): 666-672. doi: 10.19982/j.issn.1000-6621.20240523

Wang Yuanning, Du Zongmin. Research progress on CRISPR/Cas molecular diagnosis of drug-resistant Mycobacterium tuberculosis[J]. Chinese Journal of Antituberculosis, 2025, 47(5): 666-672. doi: 10.19982/j.issn.1000-6621.20240523

图/表 1

参考文献 35

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文献序号	方法	Cas蛋白	靶标基因	样品类型	扩增方法	检测耗时	检出限
[21]	/	Cas9	16s rDNA	痰液	/	2h	20CFU/ml
[22]	PC	dCas9	16s rDNA	基因组DNA	PCR	1h	5×10^-13g/ml DNA
[23]^a	CARP	Cas9	rpoB 531和526位点	基因组DNA	qRT-PCR	4h	1×10^-4 fg DNA
[24]^a	/	Cas12a RR	rpsl K43R和rpsl K88R	基因组DNA	RPA	>45min	/
[25]	TB One-Pot	Cas12b	IS6110	基因组DNA	CPA	80min	50CFU/ml
[26]	LACD	Cas12a	IS6110	痰液	LAMP	1h	50 fg DNA
[27]	/	Cas12a	IS1081或IS6110	基因组DNA	RPA	3h	2.59~1.85μg/L DNA
[28]	/	Cas12a	rpoB 531和526位点	基因组DNA	不对称RPA	1h	4.1CFU/ml
[29]	WATSON	Cas13a	IS1081或IS6110	血液	RPA	/	/
[30]	/	Cas14a	16s rDNA	基因组DNA	TWJ	1.5h	20.71 fM DNA