Conventional and real-time PCR targeting 16S ribosomal RNA for the detection of Mycobacterium tuberculosis complex
OBJECTIVE: 16S rDNA and 16S rRNA of Mycobacterium tuberculosis complex (MTC) were used as targets to develop sensitive and specific polymerase chain reactions (PCRs) to improve the diagnosis of MTC.
DESIGN: We developed conventional and real-time PCRs targeting 16S rDNA and rRNA of MTC.
RESULTS: PCRs targeting 16S rRNA had a 10−100 times lower limit of detection for M. tuberculosis than PCRs targeting 16S rDNA. The sensitivities of the 16S rDNA PCR, 16S rRNA reverse transcription PCR (RT-PCR), 16S rDNA real-time PCR and 16S rRNA real-time RT-PCR for sputum specimens were respectively 92%, 94.6%, 96% and 100%. Real-time PCR showed no cross-reactivity, but conventional PCR had cross-reactivity to M. avium, M. gastri and M. nonchromogenicum.
CONCLUSION: PCRs targeting the 16S rRNA of MTC were more sensitive than those targeting 16S rDNA; 16S rRNA real-time RT-PCR showed the highest sensitivity and specificity for MTC.
Keywords: 16S rDNA; 16S rRNA; MTC; conventional polymerase chain reaction; real-time PCR
Document Type: Research Article
Affiliations: 1: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea; Department of Biomedical Laboratory Science, Songho College, Hoengseong-gun, Korea 2: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea 3: M&D Inc, Wonju, Korea 4: Department of Microbiology, College of Medicine, Yonsei University, Seoul, Korea 5: Division of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
Publication date: 01 September 2015
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