6종의 성매개감염 병원체 검출을 위한 다중 실시간중합효소연쇄반응 검사법의 비교평가
Comparative Evaluation of Multiplex Real-Time PCR Assays for Six Pathogens of Sexually Transmitted Infections
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배경: 다중 실시간중합효소연쇄반응 검사법은 다양한 성매개감염 병원체를 동시에 검출할 수 있는 민감한 검사법이다. 본 연구에서는 다중 실시간중합효소연쇄반응을 이용하여 6종의 성매개감염 병원체를 검출하는 두 검사법을 비교 평가하였다. 방법: 성매개감염 원인 병원체 PCR 검사를 시행한 후 잔여 DNA 검체를 −70oC에 냉동하였다. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), Trichomonas vaginalis (TV)를 GeneFinder STD I (CT/NG/UU)/II (MG/MH/TV) Multiplex Real-time PCR Kits (Infopia, Korea; GeneFinder assay)와 Real-Q CT&NG/MH&TV/MG&UU Kits (BioSewoom, Korea; Real-Q assay)로 검사하였다. 두 검사법의 결과가 불일치할 경우, 다중 실시간중합효소연쇄반응을 이용하여 성매개감염 원인 병원체를 검출하는 다른 시약인 Anyplex II STI-7 Detection (Seegene, Korea)으로 검사하여, 두 개 이상의 검사법에서 양성인 결과를 진양성으로 판단하였다. 결과: 81개 검체 중 GeneFinder assay에서는 45개의 검체에서 총 63개의 병원체(16 CT, 2 NG, 6 MG, 20 MH, 18 UU, 1 TV)가 검출되었고, Real-Q assay에서는 47개의 검체에서 총 66개의 병원체(16 CT, 2 NG, 8 MG, 20 MH, 19 UU, 1 TV)가 검출되었다. 양성 검체와 음성 검체 결과를 정확하게 보고하는 것에 대한 두 검사의 일치율은 93.8%였다(Kappa=0.87). 각 병원체별 일치율은 97.5-100%였다(Kappa＞0.8). 결론: GeneFinder assay와 Real-Q assay 간에 유의한 차이를 발견할 수 없었으며, 두 다중 실시간중합효소연쇄반응 검사법 모두 임상 검사실에서 성매개감염의 병원체를 검출하는 데 유용하게 쓰일 것이다. [Ann Clin Microbiol 2017;20:1-6]
Abstract
Background: The multiplex real-time PCR assay is a sensitive test for simultaneous detection of various pathogens of sexually transmitted infections (STIs). We evaluated the performance of two multiplex real-time PCR assays for six STI pathogens. Methods: DNA samples after being used to conduct PCR for STI pathogens were stored below −70oC. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Trichomonas vaginalis (TV) were detected by multiplex real-time PCR with GeneFinder STD I (CT/NG/UU)/II (MG/MH/TV) Multiplex Real-time PCR Kits (Infopia, Korea; GeneFinder assay) and Real-Q CT&NG/ MH&TV/MG&UU Kits (BioSewoom, Korea; Real-Q assay). Discrepant results were resolved by another multiplex real-time assay, Anyplex II STI-7 Detection (Seegene, Korea). Any two positive results for the assays were considered true positive. Results: Among 81 samples, the GeneFinder assay detected 63 pathogens from 45 cases (16 CT, 2 NG, 6 MG, 20 MH, 18 UU, and 1 TV) and Real-Q assay detected 66 pathogens from 47 cases (16 CT, 2 NG, 8 MG, 20 MH, 19 UU, and 1 TV). For the results of positive cases and negative cases, the overall concordance rate between the two multiplex real-time assays was 93.8% (Kappa=0.87). For each pathogen, the agreement rates of the two assays ranged from 97.5 to 100% (Kappa＞0.8). Conclusion: There was no significant difference between the results of GeneFinder assay and Real-Q assay. Both multiplex real-time PCR assays can be useful methods for the detection of STI pathogens in clinical laboratories. (Ann Clin Microbiol 2017;20:- 6)
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