本研究将甜瓜黄斑病毒(Melon yellow spot virus,MYSV)外壳蛋白基因克隆到原核表达载体pET32a(+)中,利用大肠杆菌系统表达该蛋白,免疫家兔制备其多克隆抗体。本文通过多克隆抗体,建立了间接ELISA(ID-ELISA)、免疫捕获PCR(IC-RT-PCR)、Western blot及dot-ELISA的MYSV检测方法,以上各种方法均能特异的检测MYSV。检测结果表明:ID-ELISA方法检测效价大于1∶6 400;Western blot检测灵敏度达到1∶320(W/V,g·mL-1);Dot-ELISA检测灵敏度达到1∶80(W/V,g·mL-1),且用牙签等非实验室工具便能检测该病毒,能高效地应用于田间样品的检测。 In this study, the coat protein gene of Melon yellow spot virus (MYSV) was cloned into the prokaryotic expression vector pET32a (+). The protein was expressed in E. coli and the rabbit was immunized to prepare its polyclonal antibody. In this paper, MYSV detection methods of indirect ELISA (ID-ELISA), immunocapture PCR (IC-RT-PCR), Western blot and dot-ELISA were established by polyclonal antibody. The detection results showed that the detection titer of ID-ELISA was greater than 1: 400 and the sensitivity of Western blot was 1: 320 (W / V, g · mL-1) , g · mL-1), and the non-laboratory tools such as toothpicks can detect the virus, which can be effectively applied to the detection of the field samples.