侵染观赏百合的黄瓜花叶病毒血清学检测方法研究

采用摩擦接种和PEG沉淀法对侵染观赏百合的黄瓜花叶病毒(CMV)进行了提纯,提纯病毒具有典型的核蛋白吸收峰曲线,D260 nm/D280 nm=1.236;利用提纯的CMV免疫大白兔制备抗血清,经微量沉淀法测定,其效价为1∶1024;用硫酸铵沉淀法提纯IgG,制备了辣根过氧化物酶标抗体,利用自制抗血清对DAS-ELISA、间接-ELISA和DIBA-ELISA检测侵染观赏百合的CMV灵敏度和检测范围进行了比较试验,结果表明DAS-ELISA检测的灵敏度较高,是间接-ELISA和DIBA-ELISA灵敏度的4倍;间接-ELISA和DIBA-ELISA的检测范围比DSA-ELISA宽.对21个观赏百合样品的CMV检测结果表明,DIBA-ELISA的检出率为66.67%,间接-ELISA的检出率为38.09%,二者的检出率分别比DAS-ELISA的检出率(23.81%)提高了42.86%和14.28%.

Serological detection methods of ornamental lily infected with cucumber mosaic virus

Cucumber mosaic virus(CMV) was purified from leaves of infecting lily plant(Phaseolus vulgaris) inoculated artificially,employed the method of differential centrifugation in combination with PEG6000.The purified CMV particles had typical nuclear protein absorb curve,the ratio of D260 nm to D280 nm was 1.236.The purified CMV preparation was injected into a rabbit to produce an antiserum.The titer of antiserum were 1∶1024 by miroprecipitation test.IgG of CMV-antiserum was purified by ammonium sulfate precipitate method and the IgG was labeled by Horseradish Peroxidase(IgG-HRP).ELISA-CMV detection system was also established.The sensibility of DAS-ELISA detection CMV was the highest,it was four times as high as ones of I-ELISA and DIBA-ELISA.The range of detection CMV by I-ELISA and DIBA-ELISA were wider than one by DAS-ELISA.The rate of DIBA-ELISA and I-ELISA detection CMV were higher than that of DAS-ELISA,increased 42.86 % and 14.28 % respectively.