title: 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.
abstract: BACKGROUND
  Recently, 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.
  
  
  METHODS
  Here, 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.
  
  
  RESULTS
  The 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].
  
  
  CONCLUSIONS
  These 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.
authors.affiliation: Department of Microbiology, Chung-Ang University College of Medicine, Seoul, 156-756, Republic of Korea. kimkj@cau.ac.kr.
authors.forename: Kijeong
authors.initials: K
authors.lastname: Kim
authors.affiliation: 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, Seoul, 110-799, Republic of Korea. rsa777@snu.ac.kr.
authors.forename: Seok-Hyun
authors.initials: SH
authors.lastname: Hong
authors.affiliation: 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, Seoul, 110-799, Republic of Korea. arukas22@snu.ac.kr.
authors.forename: Byoung-Jun
authors.initials: BJ
authors.lastname: Kim
authors.affiliation: 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, Seoul, 110-799, Republic of Korea. blast3br@naver.com.
authors.forename: Bo-Ram
authors.initials: BR
authors.lastname: Kim
authors.affiliation: 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, Seoul, 110-799, Republic of Korea. leesy8822@naver.com.
authors.forename: So-Young
authors.initials: SY
authors.lastname: Lee
authors.affiliation: 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, Seoul, 110-799, Republic of Korea. love-rksk@hanmail.net.
authors.forename: Ga-Na
authors.initials: GN
authors.lastname: Kim
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea. shimts@amc.seoul.kr.
authors.forename: Tae Sun
authors.initials: TS
authors.lastname: Shim
authors.affiliation: 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, Seoul, 110-799, Republic of Korea. yhkook@snu.ac.kr.
authors.forename: Yoon-Hoh
authors.initials: YH
authors.lastname: Kook
authors.affiliation: 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, Seoul, 110-799, Republic of Korea. kbumjoon@snu.ac.kr.
authors.forename: Bum-Joon
authors.initials: BJ
authors.lastname: Kim
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D018834:Chaperonin 60 / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D005838:Genotype; D006801:Humans; D009164:Mycobacterium Infections / Q000175:diagnosis* / Q000382:microbiology; D009170:Nontuberculous Mycobacteria / Q000145:classification* / Q000235:genetics; D020135:Peptide Nucleic Acids; D011237:Predictive Value of Tests; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D013183:Sputum / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D004269:DNA, Bacterial; D020135:Peptide Nucleic Acids; C074119:heat-shock protein 65, Mycobacterium