Detection of Apple Scar Skin Viroid by Reverse Transcription Recombinase Polymerase Amplification Assay

• Main Authors: Na-Kyeong Kim, Hyo-Jeong Lee, Tae-Ho Ryu, In-Sook Cho, Ho-Jong Ju, Rae-Dong Jeong
• Format: Article
• Language: English
• Published: Hanrimwon Publishing Company 2021-06-01
• Series: Research in Plant Disease
• Subjects: apple leaves; apple scar skin viroid; detection; reverse transcription recombinase polymerase amplification
• Online Access: http://www.online-rpd.org/upload/pdf/RPD-2021-27-2-79.pdf

The aim of the present study was to develop a sensitive and specific detection method for the rapid detection of apple scar skin viroid (ASSVd) in apple leaves. The resulting reverse transcription recombinase polymerase amplification (RT-RPA) assay can be completed in 10 min at 42oC, is 10 times more sensitive than conventional reverse transcription polymerase chain reaction, and can specifically amplify ASSVd without any cross-reactivity with other common apple viruses, including apple stem grooving virus, apple stem pitting virus, and apple chlorotic leaf spot virus. The reliability of the RT-RPA assay was assessed, and the findings suggested that it can be successfully utilized to detect ASSVd in field-collected samples. The RT-RPA assay developed in the present study provides a potentially valuable means for improving the detection of ASSVd in viroid-free certification programs, especially in resource-limited conditions.