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建立马铃薯纺锤块茎类病毒属(Pospiviroid)类病毒有效的芯片筛查技术,对该属类病毒进行筛查。分析了该属类病毒序列,得到19条具有属级鉴定特征的探针。这些探针符合GC含量在40%与60%之间、单个核苷酸含量不大于50%、无多于4个核苷酸的连续重复及不形成多于6个核苷酸的发卡结构的标准;将探针点制到玻璃片基上制备芯片。芯片探针有效性实验结果表明,菊花矮化类病毒(Chrysanthemum stunt viroid,CSVd)及番茄雄性株类病毒(Tomato planta macho viroid,TPMVd)样品杂交可获得有效信号;灵敏度实验结果表明,该芯片可检测到200 pg/μL的总RNA。该芯片可用于Pospiviroid类病毒的筛查。
To establish an effective chip screening technology of potato spindle tubervirus (Pospiviroid) and to screen this genus. The genus virus sequences were analyzed and 19 probes with genus appraisal were obtained. These probes conform to the conformation of a hairpin structure with a GC content of between 40% and 60%, a single nucleotide content of no more than 50%, no more than 4 consecutive nucleotides and a hairpin structure that does not form more than 6 nucleotides Standard; the probe point system to prepare glass chips. The results of chip probe validation showed that effective signal was obtained by hybridization of Chrysanthemum stunt viroid (CSVd) and Tomato planta macho viroid (TPMVd) samples. Sensitivity experiments showed that the chip could be Total RNA of 200 pg / μL was detected. The chip can be used for Pospiviroid virus screening.