Use of a novel multiplex probe array for rapid identification of <Emphasis Type="Italic">Mycobacterium</Emphasis> species from clinical isolates |
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Authors: | Shu-Lin Zhang Jian-Guo Shen Gwan-Han Shen Zhan-Qiang Sun Ping-Hui Xu Yi-Li Peng Zhi-Rong Yang Qun Sun |
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Institution: | (1) College of Life Sciences, Key Laboratory of Bio-resource and Bio-control, Sichuan University, 29# Wangjianglu St., Chengdu, Sichuan, 610064, China;(2) Research Center for Tuberculosis, Henan Chest Hospital, Zhengzhou, Henan, China;(3) College of Basic Education, Nanyang Institute of Technology, Nanyang, Henan, China;(4) Division of Chest Medicine, Taichung Veterans General Hospital, Taiwan, China;(5) Department of Microbiology, Henan Chest Hospital, Zhengzhou, Henan, China;(6) Department of Microbiology, Institute of Biotechnology, Zhengzhou, Henan, China |
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Abstract: | Conventional identification of mycobacteria is based on the analysis of their phenotypic and biochemical characteristics after
culture; thus this method is time-consuming, laborious, and is not always conclusive. Developing a fast and accurate method
for rapid identification of Mycobacterium species is in urgent need for early diagnosis of mycobacteriosis and effective patient management. In this study, an efficient
and affordable novel multiplex probe array which allows simultaneous identification of 15 medically important mycobacterial
species was developed. A pair of genus-specific primers and a set of genus- and species-specific probes were designed according
to the conserved and polymorphic regions of the 16S rRNA gene, internal transcribed spacer (ITS) sequence, and 23S rRNA gene
of mycobacteria. This probe array was applied for the identification of 78 clinical mycobacterial isolates recovered from
Henan, China. The results showed that the specificity and sensitivity of the probe array were 100% for both genus-specific
probe and Mycobacterium tuberculosis complex-specific probe. Among 52 isolates of nontuberculous mycobacteria, 43 isolates (82.7%) can be rapidly identified to
the species level. Genetic variability of 16S-23S rRNA gene ITS region in M. avium, M. intracellulare, M. chelonae, M. abscessus and M. fortuitum were analyzed. With the accumulation of the sequences of ITS identified and further optimization of probes, the multiplex
probe array has the potential to be developed into a practical tool for rapid and accurate identification of mycobacterial
species in clinical laboratory. |
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Keywords: | Internal transcribed spacer Isolates Mycobacterium Probe Species identification Variability |
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