A蛋白酶联免疫吸附法检测水稻条纹叶枯病毒介体昆虫灰飞虱带毒率的研究

APPLICATION OF STAPHYLOCOCCAL PROTEIN A-ENZYME LINKED IMMUNOSORBENT ASSAY (SPA-EL1SA) FOR THE DETECTION OF RICE STRIPE VIRUS (RSV)

用市售的A蛋白(SPA),兔抗RSV抗体及A蛋白酶标记物(HRP-SPA)联合组成A蛋白酶联免疫吸附法(SPA-ELISA),检测RSV纯化抗原和病株粗澄清液,其灵敏度分别为0.125μg/ml和1:10000;A蛋白的适宜包被浓度为2.5μg/ml,兔抗RSV抗体作第一抗体的适宜浓度为2.5μg/ml,作第二抗体的适宜浓度为4μg/ml,用该方法检测健康稻株澄清液和提纯液,试验结果均为阴性,检测RSV介体昆虫灰飞虱带毒率,以生物接种测定法为对照,共测2028头虫,阳性虫符合率为90.13％,阴性虫符合率为98.73％。该法灵敏,简便,特异性强,适宜检测RSV介体昆虫灰飞虱带毒率,在预测该病流行方面具有实用价值。

Serological reaction of purified RSV. Supernatant liquid of RSV-infected rice plants. Laodelphax striatellus, vecters of RSV were detected by SPA-ELISA. Data of suitable concentration of various materials using in SPA-ELISA were obtained as following: 1.minimum concentration of purified virus was 0.125μg/ml; 2.maximum dilution of supernatant liquid of infected rice plants was 1:10000; 3. Optimum coating concentration of SPA was 2.5ug/ml; 4.suitable concentration of rabbit's antibody against RSV using for the first antibody and the second antibody were 2.5μg/ml and 4μg/ml, respectively. SPA-ELISA possesses the characters of high sensitivity and specificity, the coincident rate of positive and negative insects were 90.13% and 98.7% respectively in the 2028 planthoppers (L. striatellus) test. Its simplifying process was fitted to detect the percentage of planthoppers (L. striatellus) with RSV in a population. SPA-ELISA would be useful for practical value in the epidemiolgy of RSV disease.