Rapid and specific detection of the Mycobacterium tuberculosis complex using fluorogenic probes and real-time PCR

Günter Kraus, Timothy Cleary, Nancimae Miller, Rachel Seivright, Andrea K. Young, Gina Spruill, H. James Hnatyszyn

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

A rapid and sensitive strategy for the specific identification of Mycobacterium tuberculosis (TB) was designed and evaluated using crude mycobacterial lysates. The speed of real-time polymerase chain reaction (PCR) was combined with the sensitivity of fluorogenic probes to confirm the presence of mycobacteria as well as specifically identify the presence of members of the mycobacteria tuberculosis complex (MTC) in a single-tube assay. Oligonucleotides were designed to amplify the internal transcribed spacer (ITS) from several mycobacterial species. Specific fluorogenic probes were included in the PCR reaction for the identification of TB as well as Mycobacterium bovia and Mycobacterium africanum in bacterial lysates. The combination of TB-specific fluorogenic probes with real-time PCR formed an approach determined to be fast (less than 40 min), sensitive (less than 800 copies of DNA) and reliable for the specific detection of the MTC. Our data demonstrate the use of real-time PCR and fluorogenic probes in a rapid and sensitive assay to distinguish members of the MTC from other mycobacterial species.

Original languageEnglish (US)
Pages (from-to)375-383
Number of pages9
JournalMolecular and Cellular Probes
Volume15
Issue number6
DOIs
StatePublished - Jan 1 2001

Keywords

  • Detection
  • Fluorogenic probes
  • Mycobacterium
  • Real-time PCR
  • Tuberculosis

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

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    Kraus, G., Cleary, T., Miller, N., Seivright, R., Young, A. K., Spruill, G., & Hnatyszyn, H. J. (2001). Rapid and specific detection of the Mycobacterium tuberculosis complex using fluorogenic probes and real-time PCR. Molecular and Cellular Probes, 15(6), 375-383. https://doi.org/10.1006/mcpr.2001.0385