사람호흡기바이러스검출에 대한 다중PCR과 단일PCR의 성능 비교
Comparison of the Ability of Multiplex and Singleplex PCR to Detect Human Respiratory Viruses
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배경: 다중 polymerase chain reaction (PCR)은 사용이 편리하고 민감도가 높아 국내 에서 호흡기바이러스 검출의 방법으로 흔히 사용되고 있다. 그러나 하나의 반응웰에서 여러 PCR을 동시에 진행하기 때문에 간섭의 우려가 있다. 방법: 저자들은 상용화된 바이러스를 이용하여 11종의 바이러스(coronavirus 229E, OC43; parain uenza virus 1 (PIV1), parain uenza virus 2 (PIV2), and parain uenza virus 3 (PIV3); in uenza virus A (INF A), in uenza virus B (INF B); respiratory syncytial vivirus (RSV) A), respiratory syncytial virus B (RSV B); adenovirus; rhinovirus A, B, and C)를 검출할 수 있는 다중PCR 키트(AdvanSure Respiratory virus real-time PCR kit, LG Life Sciences, Korea)의 분석민감도와 간섭 정도를 단일PCR키트와 비교하였다. 결과: 코로나바이러스 229E 및 OC43, 인플루엔자 A 및 B, RSV A 및 B, 아데노바이러스, 리노바이러스 A, B, C의 측정가능한 최소농도는 다중PCR과 단일PCR이 같았다. 그러나 PIV1, 2, 3의 최소농도는 두 검사법 간에 1 희석배수의 차이가 있었다. 하나의 반응웰에 혼합된 바이러스들의 역치주기(threshold cycle, Ct) 변화는 0.24-1.99 범위로서, 서로 영향을 주지 않는 것으로 판단되었다. 결론: 호흡기바이러스 측정을 위한 다중PCR의 분석민감도는 단일PCR과 비슷하였다. 다중PCR에서 혼합된 바이러스들 간에 의미있는 간섭은 관찰되지 않았다.
Abstract
Background: The use of the multiplex polymerase chain reaction (PCR) technique for respiratory viruses has become popular in Korea owing to its convenience and sensitivity. However, concerns remain with regard to possible interference due to multiplexing. Methods: We compared the analytical sensitivity and virus interference of a commercially available, multiplex PCR kit (AdvanSure Respiratory virus real-time PCR kit, LG Life Sciences, Korea) with that of singleplex PCR to detect 11 viruses including coronavirus 229E and OC43; parainfluenza virus 1 (PIV 1), parainfluenza virus 2 (PIV 2), and parainfluenza virus 3 (PIV 3); influenza virus A (INF A) and influenza virus B (INF B); respiratory syncytial virus A (RSV A) and respiratory syncytial virus B (RSV B); adenovirus; and rhinovirus A, B, and C. Results: The lowest detected viral concentrations of coronavirus 229E and OC43, INF A and B, RSV A and B, adenovirus, and rhinovirus A, B, and C were the same for both, multiplex and singleplex systems. However, the lowest detected viral concentrations of PIV1, 2, and 3 differed by 1 dilution factor between the two systems. Threshold cycle (Ct) values for mixed viruses within the same well were not significantly influenced by each other, where the difference between Ct values ranged from 0.24 to 1.99. Conclusions: Analytical sensitivity of multiplex PCR was comparable to that of singleplex PCR for respiratory viruses. No significant interference was observed with mixed virus samples using multiplexed PCR.
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