应用免疫捕获RT-PCR检测槟榔APV1病毒

Detection of APV1 (Areca Palm Velarivirus 1) Using Immunocapture RT-PCR

槟榔黄化病的蔓延已经严重威胁海南的槟榔种植业,其病原被鉴定为APV1 (Areca palm velarivirus1)病毒。为了提高APV1病毒检测的效率及灵敏度,本研究拟采用免疫捕获RT-PCR (IC-RT-PCR)对APV1病毒进行检测。首先制备APV1病毒外壳蛋白的单克隆抗体,然后将抗体包被PCR管,加入槟榔黄化叶片或者与APV1病毒虫媒粗提取液进行免疫捕获,最后进行RT-PCR检测。研究结果显示,IC-RT-PCR对槟榔黄化病叶片样品中APV1病毒的检测灵敏度比普通RT-PCR高2 500倍,对粉蚧样品中APV1病毒的检测灵敏度比普通RT-PCR高125倍。IC-RT-PCR避免了RNA提取的环节,检测效率提高,且检测灵敏度更高,对检测的浓度样品具有巨大技术优势。本研究结果为APV1病毒的检测提供技术支撑。

Areca yellowing disease has seriously threatened the development of betel palm plantation.Its pathogen has been identified as the APV1 virus.To improve the efficiency and sensitivity of APV1 virus detection,this study discussed the use of immunocapture RT-PCR (IC-RT-PCR) to detect APV1 virus. The monoclonalantibody APV1 virus shell protein was prepared firstly, then the antibody was coated with PCR tube, then betelnut yellowing leaf was added or immunocaptured with APV1 virus insect-borne crude extract, and finally RT-PCRdetection was carried out. The results of this study showed that the detection sensitivity of IC-RT-PCR for APV1virus in betel nut yellowing disease leaf samples was 2 500 times higher than that of ordinary RT-PCR, and thedetection sensitivity of APV1 virus in mealybug samples was 125 times higher than that of ordinary RT-PCR.IC-RT-PCR avoids the link of RNA extraction and improves the detection efficiency;and the detection sensitivityis higher, and it has great technical advantages for the concentration of samples to be detected. The results of thisstudy provide technical support for the detection of APV1 virus.