Expert consensus on molecular biology diagnosis of nontuberculous mycobacteria diseases

doi:10.19982/j.issn.1000-6621.20250225

Abstract

Abstract:

The incidence of nontuberculous mycobacteria (NTM) disease has been on the rise. Traditional methods such as acid-fast bacilli staining and mycobacterial culture suffer from limitations including low sensitivity, being time-consuming, and inability to identify specific species. In contrast, molecular biology detection technologies have demonstrated unique advantages and great application prospects in the diagnosis of NTM diseases and precise species identification. However, there is lack of systematic and comprehensive guideline for the molecular diagnosis of NTM disease in China. In order to standardize the clinical application of molecular diagnostic techniques for NTM diseases and enhance the diagnosis and treatment capabilities of medical professionals, the Nontuberculous Mycobacterial Diseases Branch of Chinese Antituberculosis Association organized a writing group composed of domestic experts in the fields of NTM diseases, infectious diseases, microbiology, and methodology. Through systematic literature reviews, multiple rounds of online and offline discussions, and questionnaire surveys, the group evaluated the respective merits, limitations, and implementation status of current molecular detection technologies, discussed key issues in molecular diagnosis of NTM diseases, and ultimately formulated this consensus after iterative revisions through multiple consensus conferences. Focusing on core issues such as molecular targets for NTM disease diagnosis, molecular biological diagnostic techniques, and standardized interpretation protocols, the consensus formulated 11 evidence-based recommendations. It systematically establishes a technical framework and clinical application pathway for molecular biological diagnosis of NTM diseases, addressing core challenges like technical fragmentation and inconsistent interpretation standards. It provides actionable guidance for clinicians, clinical laboratories, and public health departments.

Key words: Mycobacterium infections, Molecular biology, Diagnosis, differential, Consensus development conference

CLC Number:

R378.91
R516

Nontuberculous Mycobacterial Diseases Branch of Chinese Antituberculosis Association. Expert consensus on molecular biology diagnosis of nontuberculous mycobacteria diseases[J]. Chinese Journal of Antituberculosis, 2025, 47(8): 961-975. doi: 10.19982/j.issn.1000-6621.20250225

Figures/Tables 3

References 163

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靶标	推荐鉴定菌种	缺陷	备注
16S rRNA	鉴定临床常见非结核分枝杆菌,如鸟分枝杆菌复合群、脓肿分枝杆菌复合群、偶发分枝杆菌复合群等,以及戈登分枝杆菌、蟾蜍分枝杆菌、玛尔摩分枝杆菌、瘰疬分枝杆菌、草分枝杆菌、耻垢分枝杆菌	不能区分堪萨斯和胃分枝杆菌、海分枝杆菌和溃疡分枝杆菌、龟分枝杆菌和脓肿分枝杆菌,对鸟分枝杆菌复合群、脓肿分枝杆菌复合群、偶发分枝杆菌复合群亚种的鉴定能力弱	现有数据库最完整
hsp65	常见非结核分枝杆菌的鉴定,如鸟分枝杆菌复合群和脓肿分枝杆菌复合群及亚种,堪萨斯分枝杆菌、胃分枝杆菌、海分枝杆菌、溃疡分枝杆菌、龟分枝杆菌、瘰疬分枝杆菌、耻垢分枝杆菌、草分枝杆菌等	对偶发分枝杆菌复合群亚种和猿分枝杆菌的鉴定能力弱	鉴别能力优于其他靶标
rpoB	常见非结核分枝杆菌的鉴定,如脓肿分枝杆菌复合群及亚种,鸟分枝杆菌、胞内分枝杆菌、龟分枝杆菌、堪萨斯分枝杆菌、胃分枝杆菌、海分枝杆菌、溃疡分枝杆菌等	对土分枝杆菌的鉴定能力弱	同时检测利福平耐药性
ITS	常见缓慢生长型分枝杆菌的鉴定,如鸟分枝杆菌复合群及亚种,胃分枝杆菌、堪萨斯分枝杆菌、瘰疬分枝杆菌、戈登分枝杆菌、玛尔摩分枝杆菌和苏尔佳分枝杆菌	对快速生长型分枝杆菌鉴定能力弱	通常作为16S rRNA的补充

技术名称	优势	缺陷
实时荧光PCR	可直接检测临床样本,应用广泛,敏感度和特异度较高	适用于初筛,部分技术不能进行非结核分枝杆菌的种属鉴定
荧光PCR熔解曲线	可直接检测临床样本,方便快捷,准确率高,经济成本低	仅能鉴定试剂盒内特定的临床常见菌种
一代测序	敏感度和特异度高,是目前非结核分枝杆菌菌种鉴定的“金标准”	耗时,操作复杂伴一定的检测污染风险
高通量测序	高通量,靶向测序可检测到预设的耐药相关基因的突变类型;宏基因组测序可同时检测包括非结核分枝杆菌在内的多种病原微生物,对混合感染有优势;全基因组测序可获得全部基因信息,有助于发现新的非结核分枝杆菌菌种	经济成本高,对样本、设备和结果解读的要求高;全基因组测序不能直接检测临床样本,耗时长,不适合大规模应用
核酸飞行时间质谱	高通量,敏感度和特异度高,可以检测分枝杆菌的混合感染和耐药相关基因的突变类型	经济成本较高,操作较为复杂
线性探针	价格低廉,敏感度和特异度较高	需要专门仪器,操作繁琐
PCR反向斑点杂交	可直接检测临床样本,敏感度和特异度较高	自动化程度偏低
基因芯片	通量较高,简单快速,敏感度和特异度较高	需PCR后处理,容易造成污染

器官与部位	常见感染菌种
肺	鸟分枝杆菌复合群、脓肿分枝杆菌复合群、堪萨斯分枝杆菌、蟾蜍分枝杆菌等
皮肤/软组织	海分枝杆菌、脓肿分枝杆菌复合群、龟分枝杆菌、偶发分枝杆菌复合群和溃疡分枝杆菌等
淋巴结	鸟分枝杆菌复合群、嗜血分枝杆菌、瘰疬分枝杆菌、玛尔摩分枝杆菌等
播散性非结核分枝杆菌病	鸟分枝杆菌复合群、龟分枝杆菌、嗜血分枝杆菌和堪萨斯分枝杆菌等
角膜/脉络膜	偶发分枝杆菌复合群、龟分枝杆菌和脓肿分枝杆菌复合群等
骨关节	海分枝杆菌、鸟分枝杆菌复合群、脓肿分枝杆菌复合群、偶发分枝杆菌复合群和龟分枝杆菌等
骨髓	脓肿分枝杆菌复合群、偶发分枝杆菌复合群和龟分枝杆菌等
牙龈	偶发分枝杆菌复合群和龟分枝杆菌
人工心脏/瓣膜置换术后	奇美拉分枝杆菌、偶发分枝杆菌复合群、龟分枝杆菌和脓肿分枝杆菌复合群等
实体器官移植术后	鸟分枝杆菌复合群