Establishment of detection method for Apple scar skin viroid(ASSVd) by Real-Time PCR

【Objective】To establish a sensitive method to detect ASSVd.【Method】According to RNA sequence of ASSVd from Gen Bank,a pair of specific primers ASSVd-F and ASSVd-R,were designed to detect ASSVd with real-time PCR. The specificity,sensitivity and applicability of the method were validated.【Result】The results showed that the template concentration detected by real- time PCR could reach 1×10-6,while it only got to 1×10-4by conventional RT-PCR,therefore the sensitivity of real-time PCR was 100 times higher than the later;Primers with strong specificity could assay ASSVd specially,while apple chlorotic leaf spot virus(ACLSV),apple stem grooving virus(ASGV) and apple stem pitting virus(ASPV) could not be detected;With the wide applicability,it could detect ASSVd from different parts of plants.【Conclusion】The established method can detect ASSVd sensitively,and it has the advantages of strong specificity and wide applicability.