自贡市非结核分枝杆菌临床分离株的菌种鉴定结果分析

目的 对自贡市35株疑似非结核分枝杆菌（NTM）临床分离株进行菌种鉴定。方法 从自贡市各区（县）及市结核病防治获得的临床分枝杆菌分离株，通过罗氏培养基（L-J）、对硝基苯甲酸（PNB）和噻吩-2-羧酸肼（TCH）鉴别培养基初步鉴定为NTM，再经多位点PCR方法确定为NTM后，对rpoB、hsp64及its基因进行测序及分析鉴定。结果 35株疑似NTM临床分离株，通过初步鉴别，多位点PCR，rpoB、hsp64及its基因测序及分析鉴别出32株NTM，分别为脓肿分枝杆菌（M. abscessus）11株、胞内分枝杆菌（M. intracellulare）7株、堪萨斯分枝杆菌（M. kansasii）5株、鸟分枝杆菌（M. avium）3株、脓毒分枝杆菌（M. septicum）1株、马赛分枝杆菌（M. masseillense）1株、戈登分枝杆菌（M. gordonae）2株、副瘰疬分枝杆菌（M. parascrofulaceum）1株和Mycobacterium peregrinum 1株。结论 抗酸染色阳性、PNB/TCH鉴定为NTM的临床分离株中包含不同种类的NTM，自贡市NTM肺病的病原种类较多，以快生长脓肿分枝杆菌为主，其次为慢生长不产色菌的胞内分枝杆菌和缓慢生长光产色菌的堪萨斯分枝杆菌。实验室应开展进一步菌种鉴定，以指导临床正确诊断和合理治疗。

Identification of clinical isolates of non-tuberculous mycobacteria in Zigong city

Objective To identify the 35 clinical suspected non-tuberculous mycobacteria (NTM) strains isolated from Zigong.Sichuan province.Methods The positive cultures from the centers for tuberculosis prevention and treatment in Zigong,were firstly identified by Roche (L-J),Nitrobenzene acid (PNB) and Thiophene-2-carboxylic acid hydrazine (TCH) culture medium.And then the NTM strains confirmed by multi-loci Polymerase chain reaction (PCR) were identified again by gene sequencing and analysis of rpoB,hsp65 and its.Results A total of 32 NTM strains were identified from the 35 clinical isolates,including 11 strains of Mycobacterium abscessus,7 strains of Mycobacterium intracellulare,5 strains of Mycobacterium kansasii,3 strains of Mycobacterium avium,1 strain of Mycobacterium septicum,1 strain of Mycobacterium masseillense,2 strain of Mycobacterium gordonae,1 strain of Mycobacterium parascrofulaceum and 1 strain of Mycobacterium peregrinum.Conclusion The species of MOTT identified by Zeihl-Neelsen stain and PNB/TCH culture in Zigong varied.Rapid growing Mycobacterium abscess was predominant,followed by the slow growing non-chromogenic Mycobacterium intracellulare and chromogenic Mycobacterium kansasii.It is necessary to conduct further identification of NTM to guide clinical diagnosis and treatment.