Real-Time PCR과 일반 PCR을 이용한 Cytomegalovirus 정성검사의 비교 평가
Comparison Cytomegalovirus Qualitative Assay Using Real-Time PCR and Conventional PCR
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배경: Cytomegalovirus (CMV) 감염은 면역력이 저하되어 있는 환자의 이환과 사망의 중요한 원인이다. 이 연구에서는CMV 검출을 위해 최근에 개발된 Real-Q Cytomegalovirus Kit (Biosewoom Inc., Korea)와 기존 제품인 PANAmPCR CMV Detection Kit (Panagene Inc., Korea)를 비교 평가하였다. 방법: 세브란스 병원에 2011년 10월에 의뢰된 CMV 정성 PCR 검사 300검체(전혈 검체: 262개, 소변 검체: 37개, 뇌척수액검체: 1개)를 대상으로 분석하였다. Real-time PCR은 Real-Q Cytomegalovirus Kit를 이용하였고 일반 PCR은 PANAmPCR CMV Detection Kit를 이용하여 CMV의 검출 부위를 증폭 시킨 후 결과를 해석하였다. 두 방법 간의 차이를 비교하고불일치 결과를 보이는 검체는 염기서열검사를 시행하였다. 결과: Real-time PCR과 일반 PCR 모두 양성률은 25.3% (76/300)였고 두 방법간의 kappa coefficients (K)는 0.96 (95% 신뢰구간, 0.93-1.00)이었다. Real-Q Cytomegalovirus Kit와 PANAmPCR CMV Detection Kit 간의 일치율은 98.7% (296/300)였고이 중, 4개의 검체에서 불일치한 결과가 나타났다. 두 방법 간의 동일한 결과를 보인 296 검체와 염기서열검사 및 환자의임상증상을 토대로 판단한 4 검체의 결과를 참값으로 할 때, 새로이 평가하고자 하는 Real-Q Cytomegalovirus Kit는 97.4% (95% 신뢰구간, 93.9-100.0%)의 민감도와 100.0% (95% 신뢰구간, 100.0-100.0%)의 특이도, 100.0% (95% 신뢰구간, 100.0- 100.0%)의 양성예측도, 99.1% (95% 신뢰구간, 97.0-100.0%)의 음성예측도를 보였다. 결론: 최근 개발된 Real-Q Cytomegalovirus Kit는 우수한 민감도와 특이도를 보였고, 기존의 일반 PCR 방법과도 높은 일치도를 보였다. 또한, real-time PCR 방법은 일반 PCR 방법의 전기영동 단계가 필요하지 않아 검사 소요 시간을 단축시킬수 있다.
Abstract
Background: Cytomegalovirus (CMV) infection is a major cause of morbidity and mortality in immunocompromised patients. We compared the abilities of the recently developed Real-Q Cytomegalovirus Kit (Biosewoom Inc., Korea) and the previously used PANA mPCR CMV Detection Kit (Panagene Inc., Korea) to detect CMV. Methods: We analyzed 300 samples (whole blood: 262, urine: 37, CSF: 1) submitted for qualitative CMV PCR testing during October 2011 at Yonsei University College of Medicine Severance Hospital. real-time PCR was performed with a Real-Q Cytomegalovirus Kit and conventional PCR was conducted with a PANAmPCR CMV Detection Kit. Results: The positive rates of both real-time PCR and conventional PCR were 25.3% (76/300), and the kappa coefficient (K) was 0.96 (95% confidence interval (CI), 0.93-1.00). The concordance rate of the Real-Q Cytomegalovirus Kit and the PANAmPCR CMV Detection Kit was 98.7% (296/300), and four out of 300 samples showed discordant results. If the concordant results of 296 samples and the four results confirmed by direct sequencing were assumed to be true, the sensitivity and specificity of the Real-Q Cytomegalovirus Kit were 97.4% (95% CI, 93.8-100.0%) and 99.1% (95% CI, 97.9-100.0%), respectively. Conclusion: The recently developed Real-Q Cytomegalovirus Kit showed excellent sensitivity and specificity, and had a high concordance rate with the previously established PANAmPCR CMV Detection Kit, which uses conventional PCR. Furthermore, real-time PCR could decrease the test time, as the electrophoresis step required for conventional PCR is not required for real-time PCR.
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