Evaluation of GenoType Mycobacterium CM for the rapid identification of non-tuberculous mycobacteria

doi:10.19871/j.cnki.xfcrbzz.2022.02.010

Abstract

Abstract: Objective To study and evaluate the GenoType Mycobacterium CM for rapid identification of NTM. Method A total of 3420 Mycobacterium positive cultures were collected from 2017 to 2018 in Shenzhen Center for Chronic Disease Control, Guangming District People's Hospital and Yantian District People's Hospital. After initial detection of GenoType MTBDRplus and elimination of the duplicate samples of patients, 127 cultures were screened out for further NTM identification through GenoType Mycobacterium CM and PCR sequencing (16S rRNA and ITS). The PCR sequencing results were used as the "gold standard" to evaluate the results of GenoType Mycobacterium CM. Result Among 127 NTM strains, there were 47 M.abscessus strains (37.0%), 20 M.fortuitum strains (15.8%), 18 M.intracellularstrains (14.2%), 11 M.gordonae strains (8.7%), 8 M.kansasii strains (6.3%), 7 M.avium strains (5.5%), 6 M.lentiflavum strains (4.7%), and 2 M.chimaera strains (1.6%), 1 M.xenopi strain (0.8%), 1 M.scrofulaceum strain (0.8%), 1 M.peregrinum strain (0.8%), 1 M.terraestrain (0.8%), 1 M.cosmeticum strain (0.8%), 1 M.kumamotonense strain (0.8%) and 1 M.dioxanotrophicus strain (0.8%). Compared with sequencing results, all 127 strains of NTM were detected by GenoType Mycobacterium CM, and 112 strains (112/127, 88.2%) could be accurately identified, 2 strains (2/127, 1.6%) could not be distinguished as M.intracellular or M.intracellular, 12 strains (12/127, 9.4%) were beyond the identification range of the kit, and 1 M.peregrinum strain (1/127, 0.8%) was incorrectly identified. Conclusion GenoType Mycobacterium CM can quickly and accurately identify common NTM, but its scope of strain identification is limited and can not fully meet the needs of clinical NTM identification.

Key words: Non-tuberculous mycobacterium, Line probe, Gene sequencing, Identification

Zhu Yumei, Zhong Yuquan, Li Jinli, Wu Likai, Wang Feng. Evaluation of GenoType Mycobacterium CM for the rapid identification of non-tuberculous mycobacteria[J]. Electronic Journal of Emerging Infectious Diseases, 2022, 7(2): 47-51.

References

[1] 中华医学会结核病学分会. 非结核分枝杆菌病诊断与治疗指南(2020年版)[J]. 中华结核和呼吸杂志, 2020, 43(11):918-946.
[2] 中华医学会结核病学分会. 分枝杆菌菌种中文译名原则专家共识编写组. 分枝杆菌菌种中文译名原则专家共识[J]. 中华结核和呼吸杂志, 2018, 41(7):522-528.
[3] BASILLE D, JOUNIEAUX V, ANDRÉJAK C. Treatment of Other Nontuberculous Mycobacteria[J]. Semin Respir Crit Care Med, 2018, 39(3):377-382.
[4] 中华医学会结核病学分会. 非结核分枝杆菌病诊断与治疗指南(2020年版)[J]. 中华结核和呼吸杂志, 2020, 43(11):918-946.
[5] 杨松, 王乐乐, 严晓峰,等. 非结核分枝杆菌病治疗药物研究进展[J]. 中华结核和呼吸杂志, 2021, 44(1):44-49.
[6] RICHTER E, RUSCH-GERDES S, HILLEMANN D.Evaluation of the GenoType Mycobacterium Assay for identification of mycobacterial species from cultures[J]. J Clin Microbiol, 2006, 44(5):1769-1775.
[7] 王峰, 朱玉梅, 桂静, 等. PCR测序和基因芯片快速鉴定分枝杆菌菌种的应用研究[J]. 中国防痨杂志, 2011, 33(11):713-717.
[8] 全国第五次结核病流行病学抽样调查技术指导组, 全国第五次结核病流行病学抽样调查办公室. 2010年全国第五次结核病流行病学抽样调查报告[J]. 中国防痨杂志, 2012, 34(8):485-508.
[9] 易俊莉, 杨新宇, 张洁,等. 三种检测技术鉴别结核分枝杆菌复合群与非结核分枝杆菌的效能评价[J]. 结核与肺部疾病杂志, 2020, 1(4):240-244.
[10] 黄明翔. 非结核分枝杆菌病的实验室诊断技术进展[J]. 中国防痨杂志, 2013, 35(7):538-541.
[11] LEE AS, JELFS P, SINTCHENKO V, et al.Identification of non-tuberculous mycobacteria: utility of the GenoType Mycobacterium CM/AS assay compared with HPLC and 16S rRNA gene sequencing[J]. J Med Microbiol, 2009, 58(7):900-904.
[12] SUBEDI S, KONG F, JELFS P, et al.16S-23S Internal Transcribed Spacer Region PCR and Sequencer-Based Capillary Gel Electrophoresis has Potential as an Alternative to High Performance Liquid Chromatography for Identification of Slowly Growing Nontuberculous Mycobacteria[J]. Plos One, 2016, 11(10):e0164138.
[13] KIM BJ, HONG SK, LEE KH, et al.Differential Identification of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria by Duplex PCR Assay Using the RNA Polymerase Gene (rpoB)[J]. J Clin Microbiol, 2004, 42(3):1308-1302.
[14] MCNABB A, EISLER D, ADIE K, et al.Assessment of partial sequencing of the 65-kilodalton heat shock protein gene (hsp65) for routine identification of Mycobacterium species isolated from clinical sources.[J]. J Clin Microbiol, 2004, 42(7): 3000-3011.
[15] 李爱芳, 谈小文, 崔晓利, 等. 荧光PCR熔解曲线法在非结核分枝杆菌菌种鉴定中的应用价值[J]. 中国防痨杂志, 2021, 43(7):664-669.
[16] 金法祥, 王华钧, 张伟阳,等. 临床分离非结核分枝杆菌的菌种分布特征分析[J]. 中华医院感染学杂志, 2017, 27(7):1480-1486.
[17] 林建, 赵永, 戴志松, 等. 福建省261株非结核分枝杆菌的菌种鉴定分析[J]. 中国防痨杂志, 2021, 43(6):590-595.
[18] 高文凤, 龙波, 杨筠, 等. 四川省46株疑似非结核分枝杆菌临床分离株菌种鉴定结果分析[J]. 寄生虫病与感染性疾病, 2019, 17(1):49-51.
[19] 刘佳文, 吕红艳, 丁北川, 等. 129株非结核分枝杆菌采用两种分子检测技术行菌种鉴定的结果分析[J]. 中国防痨杂志, 2019, 41(9):999-1004.
[20] 刘东鑫, 郑惠文, 贺文从, 等. 广州市非结核分枝杆菌临床分离株菌种鉴定及药物敏感性试验结果分析[J]. 中国防痨杂志, 2019, 41(5):534-540.
[21] 刘敏, 李明俊, 余庆, 等. 重庆市HIV与NTM双重感染患者的菌种分布及耐药情况[J]. 中国防痨杂志, 2021, 43(5):501-505.
[22] 金法祥, 陶学芳, 王华钧, 等. 2017年绍兴地区106株非结核分枝杆菌菌种鉴定结果分析[J]. 中国防痨杂志, 2019, 41(11)1179-1183.
[23] KOHLER, PHILIPP, HASSE, et al. Prolonged Outbreak of Mycobacterium chimaera Infection After Open-Chest Heart Surgery[J]. Clin Infect Dis, 2015, 61(1):67-75.
[24] VARGHESE B, ENANI M, SHOUKRI M, et al.Emergence of Rare Species of Nontuberculous Mycobacteria as Potential Pathogens in Saudi Arabian Clinical Setting[J]. PLOS Negl Trop Dis, 2017, 11(1):e0005288.
[25] COOKSEY RC.Mycobacterium cosmeticum sp. nov. a novel rapidly growing species isolated from a cosmetic infection and from a nail salon[J]. Int J Syst Evol Microbiol, 2004, 54(6):2385-2391.
[26] 谢丽,杨争,尤信信,等. 2018年深圳市学校结核病临床分离株的MIRU-VNTR基因分型特点及其传播分析[J/CD]. 新发传染病电子杂志,2021, 6(3):215-219.
[27] 谭卫国,管红云,吴清芳,等. 流动人口结核病防控创新策略与实施效果[J/CD].新发传染病电子杂志,2020,5(3):145-149.