基于RNA杂交的马铃薯纺锤块茎类病毒检测芯片

报道了一种检测植物类病毒RNA的新方法——RNA杂交芯片技术, 即将cDNA芯片技术与RNA斑点杂交技术相结合, 将马铃薯样品的总RNA直接固定在玻片上, 用荧光标记制备检测马铃薯纺锤块茎类病毒(PSTVd)的特异探针, 探针与芯片杂交后分析杂交信号以确定相应的样品有无PSTVd侵染. 参照膜杂交的方法, 确定了RNA芯片的制备条件, 并用以检测了马铃薯样品的PSTVd侵染情况, 检测结果与RT-PCR结果相符, 阳性产物经克隆测序证实为PSTVd.

Detection of Potato Spindle Tuber Viroid Using RNA Hybridized Chips

Potato diseases caused by potato spindle tuber viroid(PSTVd) bring about substantial losses in agricultural production.So developing a high throughput,rapid,sensitive and specific detection method is ne-cessary. This article reports a new method to detect plant viroidRNA glass slide hybridization.This me-thod combines dot blot hybridization technique with cDNA microarray technique.We could spot a number of RNA samples extracted from new leaves of potatoes onto a silanated glass slide directly and then perform the hybridization on the slide with Cy5-labeled probe of PSTVd prepared by PCR.Analyzing the results of hybri-dization, we could ascertain which sample is infected by PSTVd.The factors affecting the results were optimized and used to test six potato samples with PSTVd-specific probes.The developed method was used to detect two samples which infected by PSTVd.These results were in good accordance with those from the RT-PCR approach and the samples are proved to be PSTVd by cloning and sequencing.So it is an accurate and reliable method for detecting plant viroid and the first report for the diagnosis of PSTVd using dot blot hybridization and RNA glass side hybridization with fluorescently labeled probe.