两种茄科蔬菜病毒检测方法的研究

Studies on methods for detection of two Solanaceous vegetable viruses

本文研究应用三种酶联免疫吸附测定法(ELISA),检测为害茄科蔬菜的两种主要病毒,即烟草花叶病毒(TMV)和黄瓜花叶病毒(CMV)。其中双抗体夹心法(DAS-ELISA)和A蛋白间接夹心法(PAS-FLISA)可检测TMV提纯病毒0.1μg/ml与0.01μg/ml,CMV0.2μg/ml与0.008μg/ml。田间病株样品检测结果与生物测定对照结果相符。亲和素一生物素复合物间接夹心法(ABC-ELISA)可检测CMV提纯病毒0.0016μg/ml,病烟汁为1:12800,并能检测带毒蚜群体内的病毒。此法所用试剂的工作浓度为,CMV的兔抗血清(RAV-IgG)5μg/ml,鼠腹水抗体(MAV-Ab)1:500,生物素化羊抗鼠IgG(b-SAMG)1:2000,亲和素与生物素化过氧化物酶复合试剂(ABC)1:300。

The paper studies on three forms of emzyme-linked immunosorbend assay (ELISA) methods to detect two main causal viruses of solanaceous vegetable virus diseases, i.e. to bacco mosaic virus (TMV) and cucumber mosaic virus (CMV). The direct double antibody sandwich form of ELISA (DAS-ELISA) and protein A indirect samdwich ELISA (PAS-ELISA) were able to detect the lowest comcentrations of TMV purified preparation at 0.1μg/ml, 0.01μg/ml, respectively. and that of CMV purified preparation at 0.2μg/ml, 0.008μg/ml, respectively. Results of detection with ELISA methods for field virus-infected plants conformed to the results of them with bioassay. A new system of avidin-biotin-peroxidease complex indirect sandwich ELISA (ABC-ELISA) enable to detect the lowest comcentration of CMV purified preparation at 0.0016μg/ml and the highest dilution of CMV-infected tobacco sap at 1:12800. Working concentration of rabbit antiserun of CMV ( RAV-IgG) . mouse ascitic fluid antibldy (MAV-Ab),biotin-labeled sheep antibody of mouse (b-SAMG) and avidin-biotin-peroxidase complex (ABC) used for ABC-ELISA method were 5μg/ml, 1: 500 dilution, 1 : 2000 dilution and 1 : 300 dilution.