Biomedical Science Letters (대한의생명과학회지)

The Korean Society for Biomedical Laboratory Sciences (대한의생명과학회)
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Development of PCR-microplate Hybridization Assay for Detection of Mycobacterium tuberculosis

  • Lee, In-Soo (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Cho, Een-Jin (International Tuberculosis Research Center) ;
  • Cho, Sang-Nae (Department of Microbiology, Yonsei University, College of Medicine) ;
  • Kim, Tae-Ue (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Lee, Hye-Young (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University)
  • Published : 2009.12.31
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Abstract

Tuberculosis caused by Mycobacterium tuberculosis (MTB) still remains to be the most dreadful infectious disease affecting almost every country. In the present study, we developed a simple and rapid but accurate and sensitive assay method for detecting MTB using microplate hybridization assay. For this, a selective region of the rpoB gene was used to design PCR primers, and MTB and Mycobacterium genus-specific probe molecules. The specificity of the assay was confirmed using fifteen different mycobacterial reference strains and twelve different non-mycobacterial reference strains, and the sensitivity was determined to be 100 fg using genomic DNA of MTB reference strain, H37Rv. Subsequently, a total of 62 sputum samples with diverse smear scores and culture positive results were used to evaluate the kit performance. In brief, the specificity and the sensitivity of the assay were 100% and 98.4%, respectively.

Keywords

References

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