Separation of Mycobacterium abscessus into subspecies or genotype level by direct application of peptide nucleic acid multi-probe- real-time PCR method into sputa samples

BMC Infectious Diseases

Research Article

Byoung-Jun Kim¹  So-Young Lee¹  Bo-Ram Kim¹  Bum-Joon Kim¹  Seok-Hyun Hong¹  Ga-Na Kim¹  Yoon-Hoh Kook¹  Kijeong Kim²  Tae Sun Shim³ 

[1] Department of Biomedical Sciences, Microbiology and Immunology, Cancer Research Institute, and Liver Research Institute, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, 110-799, Seoul, Republic of Korea;Department of Microbiology, Chung-Ang University College of Medicine, 156-756, Seoul, Republic of Korea;Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea;

关键词: Mycobacterium abscessus;    Mycobacterium massiliense;    Peptide nucleic acid (PNA);    Real time PCR;    hsp65;    Genotype;   

DOI  :  10.1186/s12879-015-1076-8

 received in 2015-05-11, accepted in 2015-07-31,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundRecently, we introduced a novel peptide nucleic acid (PNA) multi-probe real time PCR method targeting the hsp65 gene (hsp65 PNA RT-PCR) to distinguish Mycobacterium abscessus groups.MethodsHere, we evaluated the usefulness of the hsp65 PNA RT-PCR for the direct identification of the M. abscessus group at the subspecies and genotype levels from sputa samples. The method was applied to total sputa DNA from 60 different patients who were identified as having mycobacterial infections via rpoB PCR restriction analysis of the same cultures.ResultsThe hsp65 PNA RT-PCR method had higher sensitivity than the multi-probe real-time PCR assay targeting hsp65 (HMPRT-PCR) for the detection of M. abscessus from sputum [96.7 % (29/30 samples) vs. 70 % (21/30 samples); 100 % specificity].ConclusionsThese results suggest that the PNA-based method is feasible for the detection of M. abscessus members not only from cultures but also directly from sputa.

【 授权许可】

CC BY   
© Kim et al. 2015