Qian Liang1, Yuanyuan Shang1, Fengmin Huo1, Yi Xue1, Yunxu Li1, Lingling Dong1, Shanshan Li2, Yu Pang3. 1. National Clinical Laboratory on Tuberculosis, Beijing Key Laboratory on Drug-resistant Tuberculosis Research, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing, China. 2. National Clinical Laboratory on Tuberculosis, Beijing Key Laboratory on Drug-resistant Tuberculosis Research, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing, China. Electronic address: lss9011@126.com. 3. National Clinical Laboratory on Tuberculosis, Beijing Key Laboratory on Drug-resistant Tuberculosis Research, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing, China. Electronic address: pangyupound@163.com.
Abstract
BACKGROUND: The increasing pulmonary diseases are reported to be affected by mixed infection of Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM). In this study, our objective was to assess the efficiency of mycobacterial culture plus DNA sequencing to detect the mixed infections with MTB and various NTM organisms. We also aimed to investigate how efficiently GeneXpert detected MTB in mixed infections with NTM in in vitro models. METHODS: A serial of mixed infection samples was generated by combining suspensions of MTB and five NTM bacteria, respectively. The mixed suspensions were further detected with GeneXpert and liquid culture plus DNA sequencing. RESULTS: Overall, the GeneXpert assay exhibited promising capability to identify the presence of MTB at different proportions ranging from 1% to 99%. For the liquid culture, the subsequent DNA sequencing only detected the presence of NTM bacteria in the mixed samples, which the proportion of NTM ranged from 1% to 99%, including M. intracellulare, M. kansasii, M. abscessus, and M. fortuitum. For M. avium, DNA sequencing was able to identify the mixtures as M. avium infection in suspensions with no less than 10% M. avium bacteria, whereas only MTB was found in the other suspensions with less M. avium bacteria. CONCLUSIONS: Our data demonstrate that the current diagnostic algorithm cannot yield a precise detection of mixed infections with MTB and NTM bacteria. The GeneXpert assay only identify MTB in the mixed samples, while the subculture plus DNA sequencing prefers to identify the NTM species with the higher growth rate. Further targeted molecular analysis by specific capture of multiple loci of mycobacterial species from specimens is urgently required to solve this diagnostic dilemma.
BACKGROUND: The increasing pulmonary diseases are reported to be affected by mixed infection of Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM). In this study, our objective was to assess the efficiency of mycobacterial culture plus DNA sequencing to detect the mixed infections with MTB and various NTM organisms. We also aimed to investigate how efficiently GeneXpert detected MTB in mixed infections with NTM in in vitro models. METHODS: A serial of mixed infection samples was generated by combining suspensions of MTB and five NTM bacteria, respectively. The mixed suspensions were further detected with GeneXpert and liquid culture plus DNA sequencing. RESULTS: Overall, the GeneXpert assay exhibited promising capability to identify the presence of MTB at different proportions ranging from 1% to 99%. For the liquid culture, the subsequent DNA sequencing only detected the presence of NTM bacteria in the mixed samples, which the proportion of NTM ranged from 1% to 99%, including M. intracellulare, M. kansasii, M. abscessus, and M. fortuitum. For M. avium, DNA sequencing was able to identify the mixtures as M. aviuminfection in suspensions with no less than 10% M. avium bacteria, whereas only MTB was found in the other suspensions with less M. avium bacteria. CONCLUSIONS: Our data demonstrate that the current diagnostic algorithm cannot yield a precise detection of mixed infections with MTB and NTM bacteria. The GeneXpert assay only identify MTB in the mixed samples, while the subculture plus DNA sequencing prefers to identify the NTM species with the higher growth rate. Further targeted molecular analysis by specific capture of multiple loci of mycobacterial species from specimens is urgently required to solve this diagnostic dilemma.