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Use of a novel multiplex probe array for rapid identification of Mycobacterium species from clinical isolates

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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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Acknowledgements

This study was supported by grant 971200100 from Henan Key Scientific and Technological Project, and grant 981170835 from Henan Scientific and Technological Project, China. We thank Jie Zhang in Sichuan University for excellent technique assistance, and thank Jin-Shui Wang in South China University of Technology for helpful suggestions.

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Authors and Affiliations

  1. College of Life Sciences, Key Laboratory of Bio-resource and Bio-control, Sichuan University, 29# Wangjianglu St., Chengdu, Sichuan, 610064, China

    Shu-Lin Zhang, Zhi-Rong Yang & Qun Sun

  2. Research Center for Tuberculosis, Henan Chest Hospital, Zhengzhou, Henan, China

    Shu-Lin Zhang & Yi-Li Peng

  3. College of Basic Education, Nanyang Institute of Technology, Nanyang, Henan, China

    Jian-Guo Shen

  4. Division of Chest Medicine, Taichung Veterans General Hospital, Taiwan, China

    Gwan-Han Shen

  5. Department of Microbiology, Henan Chest Hospital, Zhengzhou, Henan, China

    Zhan-Qiang Sun

  6. Department of Microbiology, Institute of Biotechnology, Zhengzhou, Henan, China

    Ping-Hui Xu

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  1. Shu-Lin Zhang
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  2. Jian-Guo Shen
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  3. Gwan-Han Shen
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  8. Qun Sun
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Correspondence to Qun Sun.

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Zhang, SL., Shen, JG., Shen, GH. et al. Use of a novel multiplex probe array for rapid identification of Mycobacterium species from clinical isolates. World J Microbiol Biotechnol 23, 1779–1788 (2007). https://doi.org/10.1007/s11274-007-9428-1

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  • DOI: https://doi.org/10.1007/s11274-007-9428-1

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