Real-time PCR을 이용한 Epstein-Barr Virus DNA 검사 평가
Evaluation of Real-time PCR Kits for Epstein-Barr Virus DNA Assays
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배경: Epstein-Barr virus (EBV)는 전염성 단구증, 악성종양의 원인으로 밝혀져 있다. 현재 국내에서 시판되는 EBV 검사 시약 중 real-time PCR을 이용한 3종류의 시약을 비교하여 보았다. 방법: 2013년 1월부터 2014년 9월까지 연세의대 세브란스병원에 EBV PCR 검사가 의뢰된 검체 중 검체량이 충분한 300검체를 임의로 선택하였다. Artus EBV RG PCR Kit (Qiagen, Germany), AccuPower EBV Quantitative PCR Kit (Bioneer, Korea), 그리고 Real-Q EBV Kit (BioSewoom, Korea)를 비교하였다. 결과: 세 가지 시약으로 검사한 결과 정성 검사 기준으로 각 검사들의 일치율과 Kappa 계수는 86.3%, 0.69 (Artus-AccuPower), 93.3%. 0.85 (Artus-Real-Q) 그리고 92.3%, 0.83 (AccuPower-Real-Q)이었다. 세 시약 결과 간의 상관성은 유의하였는데, 상관계수 r=0.854 (Artus-AccuPower), -0.802 (Artus-Real-Q), 그리고 -0.977 (AccuPower-Real-Q)이었다. 하나라도 결과가 불일치한 42검체는 sequencing 결과를 참으로 가정하여 민감도와 특이도를 구하였을 때 0.921/0.976 (Artus), 0.902/0.965 (AccuPower), 그리고 0.967/ 1.000 (Real-Q)이었다. 결론: 세 가지 real-time PCR을 이용한 EBV 검사는 모두 민감도와 특이도가 우수하였다. 임상 검사실에서는 각자의 조건에 따라 선택하면 좋을 것이다. 그렇지만 임상적 진단과 추적 검사 시 동일한 시약을 사용하는 것이 필요하며, 앞으로 표준화된 검사 방법이 필요할 것으로 생각된다.
Abstract
Background: Epstein-Barr virus (EBV) is known to be the causative agent of infectious mononucleosis and EBV-related malignancies. In this study, we compared the results of three real-time PCR kits for EBV DNA assays. Methods: A total of 300 whole blood samples submitted for quantitative EBV PCR between January 2013 and September 2014 at Severance Hospital were included. The samples were tested by using the Artus EBV RG PCR Kit (Qiagen, Germany), AccuPower EBV Quantitative PCR Kit (Bioneer, Korea), and Real-Q EBV Kit (BioSewoom, Korea). Samples with discordant results between the three kits were confirmed by direct sequencing. Results: The result concordance rate and kappa coefficient (K) were 86.3% and 0.69 for Artus–AccuPower, 93.3% and 0.85 for Artus–Real-Q, and 92.3% and 0.83 for AccuPower–Real-Q, respectively. The correlations between the three kits were found to be significant, with a correlation coefficient of r=0.854 for Artus–AccuPower, -0.802 for Artus–Real-Q, and -0.977 for AccuPower–Real-Q, respectively (P<0.0001). If the real-time PCR concordant results of 258 samples and the direct sequencing results of 42 real-time PCR discordant samples were assumed to be true, the sensitivity/specificity values were 0.921/0.976 for Artus, 0.902/0.965 for AccuPower, and 0.967/1.000 for Real-Q. Conclusions: The three real-time PCR kits showed excellent sensitivities and specificities. All these kits would be acceptable for clinical and therapeutic management of EBV. However, some discordant results between the kits indicate the need for caution in clinical diagnosis and staging. Further implementation of standardized methodology would be needed for EBV DNA assays.
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