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J Clin Microbiol. 2019 Jan 2;57(1). pii: e01096-18. doi: 10.1128/JCM.01096-18. Print 2019 Jan.

Rapid Identification of Clinically Relevant Mycobacterium Species by Multicolor Melting Curve Analysis.

Xu Y#1,2,3, Liang B#1,2,3, Du C1,2, Tian X1,2, Cai X4, Hou Y5, Li H6, Zheng R7, Li J8, Liu Y5, Wang K5, Ammar Athar M1, Tan Y9, Li Q10,2,3.

Author information

1
Engineering Research Centre of Molecular Diagnostics, Ministry of Education, State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian, China.
2
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, Fujian, China.
3
Engineering Research Centre of Personalized Molecular Diagnostics of Xiamen, Xiamen, Fujian, China.
4
Department of Clinical Laboratory, Guangzhou Chest Hospital, Guangzhou, Guangdong, China.
5
Infectious Disease Hospital of Heilongjiang province, Haerbin, Heilongjiang, China.
6
Tuberculosis Reference Laboratory, Henan Provincial Center for Disease Control and Prevention, Zhengzhou, Henan, China.
7
Division of Tuberculosis Control and Prevention, Xiamen Center for Disease Control and Prevention, Xiamen, Fujian, China.
8
Chest Hospital of Xinjiang Uyghur Autonomous Region, Wulumuqi, Xinjiang Uyghur Autonomous Region, China.
9
Department of Clinical Laboratory, Guangzhou Chest Hospital, Guangzhou, Guangdong, China gzchtan@163.com qgli@xmu.edu.cn.
10
Engineering Research Centre of Molecular Diagnostics, Ministry of Education, State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian, China gzchtan@163.com qgli@xmu.edu.cn.
#
Contributed equally

Abstract

The sustained increase in the incidence of nontuberculous mycobacterial (NTM) infection and the difficulty in distinguishing these infections from tuberculosis constitute an urgent need for NTM species-level identification. The MeltPro Myco assay is the first diagnostic system that identifies 19 clinically relevant mycobacteria in a single reaction based on multicolor melting curve analysis run on a real-time PCR platform. The assay was comprehensively evaluated regarding its analytical and clinical performances. The MeltPro Myco assay accurately identified 51 reference mycobacterial strains to the species/genus level and showed no cross-reactivity with 16 nonmycobacterial strains. The limit of detection was 300 bacilli/ml, and 1% of the minor species was detected in the case of mixed infections. Clinical studies using 1,163 isolates collected from five geographically distinct health care units showed that the MeltPro Myco assay correctly identified 1,159 (99.7%) samples. Further testing with 94 smear-positive sputum samples showed that all samples were correctly identified. Additionally, the entire assay can be performed within 3 h. The results of this study confirmed the efficacy of this assay in the reliable identification of mycobacteria, suggesting that it might potentially be used as a screening tool in regions endemic for tuberculosis.

KEYWORDS:

melting curve analysis; nontuberculous mycobacteria; real-time PCR; species identification

PMID:
30333128
DOI:
10.1128/JCM.01096-18

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