Hoza Abubakar S, Mfinanga Sayoki G M, Rodloff Arne C, Moser Irmgard, König Brigitte
Institute of Medical Microbiology and Epidemiology of Infectious Diseases, University of Leipzig, Liebigstrasse 21, 04103, Leipzig, Germany.
Sokoine University of Agriculture, Morogoro, Tanzania.
BMC Res Notes. 2016 Feb 17;9:109. doi: 10.1186/s13104-016-1928-3.
Non-tuberculous mycobacteria (NTM) are increasingly reported worldwide associated with human disease. Defining the significance of NTM in settings with endemic tuberculosis (TB) requires the discrimination of NTM from TB in suspect patients. Correct and timely identification of NTM will impact both therapy and epidemiology of TB and TB-like diseases. The present study aimed at determining the frequency and diversity of NTM among TB suspects in northeastern Tanzania.
A cross-sectional study was conducted between November 2012 through January 2013. Seven hundred and forty-four sputum samples were collected from 372 TB suspects. Detection was done by using phenotypic, GenoType(®) Mycobacterium CM/AS kits, 16S rRNA and hsp65 gene sequencing for identification of isolates not identified by Hain kits. Binary regression model was used to analyse the predictors of NTM detection.
The prevalence of NTM was 9.7% of the mycobacterial isolates. Out of 36 patients with confirmed NTM infection, 12 were HIV infected with HIV being a significant predictor of NTM detection (P < 0.001). Co-infection with Mycobacterium tuberculosis (M. tb) was found in five patients. Twenty-eight NTM isolates were identified using GenoType(®) Mycobacterium CM/AS and eight isolates could not be identified. Identified species included M. gordonae and M. interjectum 6 (16.7%), M. intracelullare 4 (11.1%), M. avium spp. and M. fortuitum 2 (5.5%), M. kansasii, M. lentiflavum, M. simiae, M. celatum, M. marinum 1 (2.8%) each. Of isolates not identified to subspecies level, we identified M. kumamotonense (2), M. intracellulare/kansasii, M. intermedium/triplex, M. acapulcensis/flavescens, M. stomatepiae, M. colombiense and M. terrae complex (1) each using 16S rRNA sequencing. Additionally, hsp65 gene sequencing identified M. kumamotonense, M. scrofulaceum/M. avium, M. avium, M. flavescens/novocastrense, M. kumamotonense/hiberniae, M. lentiflavum, M. colombiense/M. avium and M. kumamotonense/terrae/hiberniae (1) each. Results of the 16S rRNA and hsp65 gene sequencing were concordant in three and discordant in five isolates not identified by GenoType(®) Mycobacterium CM/AS.
NTM infections may play a vital role in causing lung disease and impact management of TB in endemic settings. GenoType(®) Mycobacterium CM/AS represents a useful tool to identify clinical NTM infections. However, 16S rRNA gene sequencing should be thought for confirmatory diagnosis of the clinical isolates. Due to the complexity and inconsistence of NTM identification, we recommend diagnosis of NTM infections be centralized by strengthening and setting up quality national and regional infrastructure.
非结核分枝杆菌(NTM)在全球范围内与人类疾病相关的报道日益增多。在结核病(TB)流行地区确定NTM的意义需要在疑似患者中将NTM与TB区分开来。正确及时地鉴定NTM将影响TB及类TB疾病的治疗和流行病学。本研究旨在确定坦桑尼亚东北部TB疑似患者中NTM的频率和多样性。
于2012年11月至2013年1月进行了一项横断面研究。从372名TB疑似患者中收集了744份痰标本。采用表型、GenoType®分枝杆菌CM/AS试剂盒、16S rRNA和hsp65基因测序对Hain试剂盒未鉴定的分离株进行检测。采用二元回归模型分析NTM检测的预测因素。
NTM在分枝杆菌分离株中的患病率为9.7%。在36例确诊NTM感染的患者中,12例感染了HIV,HIV是NTM检测的一个重要预测因素(P<0.001)。5例患者同时感染了结核分枝杆菌(M.tb)。使用GenoType®分枝杆菌CM/AS鉴定了28株NTM分离株,8株分离株无法鉴定。鉴定出的菌种包括戈登分枝杆菌和中间分枝杆菌6株(16.7%)、细胞内分枝杆菌4株(11.1%)、鸟分枝杆菌复合群和偶然分枝杆菌2株(5.5%)、堪萨斯分枝杆菌、缓黄分枝杆菌、猿分枝杆菌、塞拉分枝杆菌、海分枝杆菌各1株(2.8%)。对于未鉴定到亚种水平的分离株,我们使用16S rRNA测序分别鉴定出熊本分枝杆菌(2株)、细胞内/堪萨斯分枝杆菌、中间/三联分枝杆菌、阿卡普尔科/微黄分枝杆菌、口部分枝杆菌、哥伦比亚分枝杆菌和土地分枝杆菌复合群(各1株)。此外,hsp65基因测序分别鉴定出熊本分枝杆菌、瘰疬分枝杆菌/鸟分枝杆菌、鸟分枝杆菌、微黄/新卡斯特伦分枝杆菌、熊本/冬分枝杆菌、缓黄分枝杆菌、哥伦比亚/鸟分枝杆菌和熊本/土地/冬分枝杆菌(各1株)。在GenoType®分枝杆菌CM/AS未鉴定的分离株中,16S rRNA和hsp65基因测序结果在3株中一致,在5株中不一致。
NTM感染可能在导致肺部疾病中起重要作用,并影响流行地区TB的管理。GenoType®分枝杆菌CM/AS是鉴定临床NTM感染的有用工具。然而,对于临床分离株的确诊,应考虑采用16S rRNA基因测序。由于NTM鉴定的复杂性和不一致性,我们建议通过加强和建立高质量的国家和地区基础设施将NTM感染的诊断集中化。