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An efficient alternative marker for specific identification of Mycobacterium tuberculosis

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Abstract

Rapid and accurate identification of mycobacteria to the species level is important to provide epidemiological information and to guide the appropriate treatment, especially identification of the Mycobacterium tuberculosis (MTB) which is the leading pathogen causing tuberculosis. The genetic marker named as Mycobacterium tuberculosis specific sequence 90 (mtss90) was screened by a bioinformatics software and verified by a series of experiments. To test its specificity, 266 strains of microorganisms and human cells were used for the mtss90 conventional PCR method. Moreover, the efficiency of mtss90 was evaluated by comparing 16S rDNA (Mycobacterium genus-specific), IS6110 (specific identification of MTB complex), mtp40 (MTB-specific) and PNB/TCH method (traditional bacteriology testing) in Mycobacterium strains. All MTB isolates were mtss90 positive. No amplification was observed from any other tested strains with M. microti as an exception. Compared with the traditional PNB/TCH method, the coincidence rate was 99.1 % (233/235). All of the mtss90 positive strains were IS6110 and 16S rDNA positive, indicating a 100 % coincidence rate (216/216) between mtss90 and these two genetic markers. Additionally, mtss90 had a better specificity than mtp40 in the identification of MTB. Lastly, a real-time PCR diagnostic assay was developed for the rapid identification of MTB. In conclusion, mtss90 may be an efficient alternative marker for species-specific identification of MTB and could be used for the diagnosis of tuberculosis combined with other genetic markers.

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Acknowledgments

We thank the National Institutes for Food and Drug Control, Guozhi Wang and Baowen Chen for their generous providing of the DNA of mycobacteria. This study was supported by the grants from Project Supported by the Scientific and Technological Research Program of Chongqing Municipal Education Commission (Grant No. KJ120318), the National Natural Science Foundation of China (No. 81101216), and the Natural Science Foundation of Chongqing (No. cstc2012jjA10009).

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Authors and Affiliations

  1. Department of Laboratory Medicine, College of Laboratory Medicine, Chongqing Medical University, 1 Yixueyuan Road, Yuzhong District, Chongqing, 400016, People’s Republic of China

    Jianing Zhao, Xuemei Zhang, Yujuan He, Hong Wang, Yibing Yin & Wenchun Xu

  2. Department of Clinical Laboratory, Chongqing Pulmonary Hospital, Chongqing, People’s Republic of China

    Yiwei Wang

  3. Department of Pneumatology, The First Affiliated Hospital, Chongqing Medical University, Chongqing, People’s Republic of China

    Dairong Li & Ju Cao

  4. Department of Clinical Laboratory, Beijing Geriatric Hospital, Beijing, People’s Republic of China

    Jiawen Liu

  5. Department of Clinical Laboratory, Children’s Hospital, Chongqing Medical University, Chongqing, People’s Republic of China

    Yibing Yin

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  1. Jianing Zhao
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  2. Yiwei Wang
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  3. Dairong Li
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  5. Xuemei Zhang
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  7. Hong Wang
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  8. Ju Cao
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  9. Yibing Yin
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  10. Wenchun Xu
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Correspondence to Wenchun Xu.

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Zhao, J., Wang, Y., Li, D. et al. An efficient alternative marker for specific identification of Mycobacterium tuberculosis . World J Microbiol Biotechnol 30, 2189–2197 (2014). https://doi.org/10.1007/s11274-014-1638-8

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  • DOI: https://doi.org/10.1007/s11274-014-1638-8

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