- 期刊
使用即時定量RT－PCR於瓜類褪綠黃化病毒檢驗之評估
Evaluation of a TaqMan One-Step Real-Time RT-PCR Method for the Detection of CCYV
摘要
本研究依據瓜類褪綠黃化病毒（Cucurbit chlorotic yellows virus; CCYV）之鞘蛋白（coat protein）基因設計專一性引子對，並利用real-time RT-PCR與TaqMan螢光探針，建立瓜類褪綠黃化病毒的檢測技術，同時進行田間洋香瓜和南瓜是否感染CCYV之測試。結果顯示，本研究所開發之real-time RT-PCR檢測方法的靈敏度，相較於一般RT-PCR提高約100－1,000倍，並可在感病洋香瓜和南瓜上專一性的偵測到CCYV，與其他瓜類病毒及健康之植株並無反應。本檢測方法亦運用於分析田間感染CCYV洋香瓜試驗。此real-time RT-PCR具有專一性佳及靈敏度高之優點，未來可將本技術應用於評估瓜類作物是否感染CCYV。
並列摘要
In this study, a diagnostic assay for Cucurbit chlorotic yellows virus (CCYV) using TaqManbased real-time reverse transcription polymerase chain reaction (qRT-PCR) was developed. Specific primers were designed by targeting the coat protein gene of CCYV. The sensitivity of qRT-PCR for the detection of CCYV when compared to that of RT-PCR assay was approximately 100–1,000 times higher. The assay is highly specific for CCYV only, no cross-reaction was observed on cucurbits infected with other cucurbit viruses using this assay. The technique is sensitive and accurate, and allows rapid detection and quantitation of CCYV in both field and experimental material used for the surveillance and diagnosis of CCYV.
延伸閱讀
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