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鉴于生产实践的需要,中国科学院武汉病毒研究所研究利用分子探针的技术,快速诊断柑桔裂皮病方法。分子探针的原理和方法是:制备柑桔裂皮病类病毒RNA,在高浓度盐的情况下,将环状RNA解环成线状,以多聚腺苷酸聚合酶在线状RNA的末端连上一段多聚腺苷酸。以此为模板,在逆转录酶的作用下,加入四种三磷酸脱氧核苷酸(其中一种用刮—~(32)P—ATP标记)为底物合成一条cDNA,经碱解后把标记的cDNA分离出来作为探针,以放射性检测柑桔树
In view of the needs of production practice, Wuhan Institute of Virology, Chinese Academy of Sciences, the use of molecular probe technology to quickly diagnose citrus split skin disease method. The principle and method of molecular probes are: Preparation of citrus split virus RNA, in the case of high concentrations of salt, the circular RNA is deconjugated into a linear polyadenylate polymerase in the linear RNA ends Even a polyadenylation. Using this as a template, four kinds of deoxynucleoside triphosphates (one of which is labeled with scratch- (32) P-ATP) were synthesized under the action of reverse transcriptase to synthesize a cDNA. After alkaline hydrolysis, The labeled cDNA is isolated as a probe to radioactively detect citrus trees