Annals of Laboratory Medicine

Korean Society for Laboratory Medicine (대한진단검사의학회)
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Identification of Nontuberculous Mycobacteria by Sequence Analysis of the 16S Ribosomal RNA, the Heat-shock Protein 65 and the RNA Polymerase ${\beta}$-Subunit Genes

16S Ribosomal RNA, Heat-shock Protein 65 및 RNA Polymerase ${\beta}$-Subunit 유전자 염기서열분석을 통한 비결핵 마이코박테리아의 동정

Shin, Sue;Kim, Eui-Chong;Yoon, Jong-Hyun
신수;김의종;윤종현

  • Published : 20060000
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Abstract

Background : The diagnosis of diseases caused by nontuberculous mycobacteria (NTM) is difficult, because NTM are prevalent in the environment such as soil and water, and because they have fastidious properties. In this study we investigated clinical isolates of NTM for their distribution pattern and accurate species identification. Methods : We selected presumptive NTM isolates negative for probe hybridization for M. tuberculosis complex, cultured in a third referral hospital from 21 January 2003 to 20 January 2004. Ninety seven-isolates were identified to the species level by direct sequencing of fragments of 16S rRNA, hsp65 and rpoB genes. A total of 120 isolates were studied for the distribution analysis. Results : Frequently identified NTM species were M. avium (30.8%), M. intracellulare (23.3%) and M. abscessus (18.3%). Others were M. gordonae, M. senegalense, M. fortuitum, M. peregrinum, M. kansasii, M. terrae complex, M. lentiflavum, M. chelonae, and M. szulgai. Three M. tuberculosis complex (2.5%) were also identified among the presumptive NTM isolates. The identification rate by sequencing of 16S rRNA, rpoB, and hsp65 were 65%, 82% and 87%, respectively. The hsp65 or rpoB gene was more efficient than 16S rRNA for the identification of NTM by sequencing. Conclusions : Some NTM are increasingly considered to be the causative organisms in clinical diseases. Thus, direct sequencing could be adapted to routine work of clinical laboratories for accurate identification of NTM to the species level.

배경 : 비결핵 마이코박테리아(NTM)는 환경분포균으로 환자검체에서 분리되는 경우라도 질환과 관계없는 오염균으로 생각하기 쉽고, 일반적인 미생물학적 동정방법으로는 정확한 동정이 어렵다. 이에 저자들은 임상검체에서 배양된 NTM에 대하여 염기서열분석법으로 정확하게 동정하여 균종 분포를 연구하고자 하였다. 방법 : 2003년 1월 21부터 2004년 1월 20일까지 일개 3차 의료기관의 결핵검사실에서 배양된 균주 중에서 결핵균 소식자에 음성인 NTM를 대상으로 하였다. 이들 중 집락이 10개 이상 배양되거나 10집락 미만이라도 동일 환자에서 두번 이상 배양된 97개에서 16S rRNA, hsp65 및 rpoB 유전자 서열을 분석하였고, 결핵연구원의 결과를 포함하여 120환자에서 균종 분포를 연구하였다. 결과 : 임상검체에서 배양된 NTM 균종은 M. avium, M. intracellulare, M. abscessus가 각각 30.8%, 23.3%, 18.3%를 차지하였다. 이외에 M. gordonae, M. senegalense, M. fortuitum, M. peregrinum, M. kansasii, M. terrae complex, M. lentiflavum, M. chelonae, M. szulgai가 분리되었고 MTB complex도 2.5%에서 확인되었다. 유전자별 동정 성공률은 16S rRNA, rpoB, hsp65 각 65%, 82%, 87%로써, NTM을 염기서열분석법으로 동정할 때에는 5'-16S rDNA보다는 hsp65나 rpoB를 이용하는 것이 성공률을 높일 수 있었다. 결론 : 임상 검체에서 NTM은 더 이상 오염균으로 무시할 수 있는 대상이 아니며, 염기서열분석을 이용한 마이코박테리아의 동정법은 검사실에서도 유용하게 쓰일 수 있을 것으로 기대한다.

Keywords

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