分枝杆菌对硝基苯甲酸实验及其鉴别分枝杆菌的分子机制研究进展

doi:10.19982/j.issn.1000-6621.20230191

摘要/Abstract

摘要：

分枝杆菌病是全球范围内威胁人类健康的重要传染病。其中,结核分枝杆菌(Mycobacterium tuberculosis,MTB)和非结核分枝杆菌(non-tuberculous mycobacteria,NTM)所导致的疾病在临床症状、病理表现及影像学特征等方面难以区分,但两者的耐药谱天然存在显著差异,致使两种疾病在临床处置方面显著不同,因此需要快速、准确地鉴别这两类分枝杆菌。PNB(4-nitrobenzoic acid)实验是基于分枝杆菌对对硝基苯甲酸不同的敏感性而建立的鉴别方法,由于操作简单、成本低廉、设备依赖性低,在临床上一直被广泛应用于MTB和NTM的鉴别,但其区分两类分枝杆菌的分子机制尚不十分明确。作者从PNB实验方法、已有研究的分枝杆菌PNB代谢酶,以及尚存在的问题等几个方面进行综述,以期为阐明分枝杆菌PNB耐药机制、其区分MTB与NTM的原理,以及PNB的新应用价值提供思路。

关键词: 硝基苯甲酸盐, 分枝杆菌,结核, 非结核分枝杆菌

Abstract:

Mycobacterial disease is an important infectious disease threatening human health worldwide. The diseases caused by Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM) are very similar in clinical symptoms, pathological manifestations and imaging features. However, there are inherent differences in the drug resistance spectrum between these two types of strains, resulting in significant differences in clinical treatment and management, therefore, rapid and accurate differentiation method is needed. PNB (4-nitrobenzoic acid) experiment is a selective culture method based on the different susceptibility of mycobacteria to PNB. It is widely used in clinical practice due to its simplicity, low price, and low technical requirements, but the mechanism involved in this experiment is unclear. This article reviewed the PNB experimental methods, reported researches on the PNB metabolic enzymes of mycobacteria and existing problems to provide ideas for elucidating the mechanism of mycobacteria PNB resistance, the principle of distinguishing MTB from NTM, and the new application value of PNB.

Key words: Nitrobenzenes, Mycobacterium tuberculosis, Non-tuberculous mycobacterium

中图分类号:

R378.91

李月, 夏辉, 李马超, 王瑞白. 分枝杆菌对硝基苯甲酸实验及其鉴别分枝杆菌的分子机制研究进展[J]. 中国防痨杂志, 2023, 45(10): 997-1002. doi: 10.19982/j.issn.1000-6621.20230191

Li Yue, Xia Hui, Li Machao, Wang Ruibai. 4-nitrobenzoic acid experiment of mycobacteria and research progress on its molecular mechanism of Mycobacterium differentiation[J]. Chinese Journal of Antituberculosis, 2023, 45(10): 997-1002. doi: 10.19982/j.issn.1000-6621.20230191

图/表 2

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方法名称	基质类型	培养基	PNB浓度	样本量	报告时间
传统法	固体	罗氏培养基	500μg/ml	100μl 10^-1mg/ml	28d
PNB-MGIT	液体	BACTEC MGIT 960	500μg/ml	1麦氏浊度, 1∶5稀释后接种500μl	7~10d
微量肉汤法		Middlebrook 7H9 ( 10%OADC)	200μg/ml	10^-3mg/ml,100μl接种	7~10d
PNB-7H10	固体	Middlebrook 7H10 (10%OADC)	500μg/ml	1麦氏浊度,1∶100稀释后10μl接种	3~28d
NRAp		罗氏培养基(1000μg/ml KNO₃)	500μg/ml	200μl痰液接种,1ml Griess显色	7~10d
分光光度计	代谢产物	水	0.001%	20mg/ml	8h
LC-MS		水	0.001%	20mg/ml	8h