도말 양성 임상검체에서 MolecuTech Real MTB-ID을 이용한 결핵균과 비결핵균의 검출평가
Evaluation of MolecuTech Real MTB-ID for MTB/NTM Detection Using AFB Smear-positive Direct Specimens
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초록
배경: 실시간 중합효소연쇄반응은 유전자의 증폭산물에 결합된 형광물질로부터 방출되는 형광량을 실시간으로 측정하여 분석하는 방법으로 중합효소연쇄반응 이후 전기영동, 교잡반응과 같은 추가적인 과정없이 분석이 가능하므로 기존 중합효소연쇄반응보다 오염가능성과 위음성의 가능성이 개선되고 검사 소요시간도 단축되는 장점이 있다. 본 연구에서, 국내에서 개발된 real time PCR 제품을 사용하여 임상검체에서 결핵균과 비결핵균을 검출하는 방법을 평가하고자 하였다. 방법: 민감도 평가는 삼성서울병원에서 의뢰된 155의 검체(도말, 항산균 배양 등의 검사를 통해 최종 동정결과가 항산균 염색 양성 59 결핵검체와 96 NTM 검체를 대상으로 실시하였다. 검사는 MolecuTech Real MTB-IDⓇ (M&D, 한국)으로실시하였으며, 결핵균은 IS6110-RD, 마이코박테리아는 rpoB 유전자를 각각 증폭시켜 검출하였다. 결과: 결핵균과 NTM의 양성률은 결핵균 검체에서 98.3% (58/59), NTM 검체에서 97.9% (94/96)로 각각 나타났으며 REBA Myco-IDⓇ에 의해 결핵균/NTM이 섞여있던 검체의 MTB/Myc probe Ct값을 비교해 본 결과 32.3-31.8, 35.3-33.8 그리고35.5-32.9로 Myc probe의 Ct값이 더 높게 나타난 것을 확인할 수 있었다. 결론: MolecuTech Real MTB-IDⓇ은 민감도와 특이도가 높게 나타났으며 직접검체에서 결핵균과 NTM을 빠르게 구별하는 진단에 있어서 유용하게 사용될 수 있을 것으로 판단된다.
Abstract
Background: The differentiation of Mycobacterium tuberculosis (MTB) from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy. The diagnosis of diseases caused by 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 evaluated real-time PCR based MTB/NTM detection kit for its usefulness in discrimination of MTB and NTM species. Methods: A total of one hundred fifty five sputum specimens whose AFB staining smear and culture had been positives were used for this study. Among them, 59 and 96 had been identified as MTB and NTM, respectively. DNAs obtained from sputum specimens were subjected for MolecuTech Real MTB-IDⓇ (M&D, Korea), real-time PCR-based MTB/ NTM detection kit. Subsequently, the results of MolecuTech Real MTBIDⓇ were compared with AFB staining smear and culture results. Results: The positive rate of MolecuTech Real MTBIDⓇ to detect MTB and NTM were 98.3% (58/59) and 97.9 (94/96) respectively using sputum specimens. Conclusion: For detection of MTB/NTM, the sensitive and specific of MolecuTech Real MTB-IDⓇ was comparable to those of conventional methods. Therefore, this study suggests the usefulness of real-time PCR based MolecuTech Real MTB-IDⓇ for rapid and detection of MTB/NTM from direct specimens.
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