A multiple RT-PCR assay for simultaneous detection and differentiation of latent viruses and apscarviroids in apple trees.

摘要: Apple chlorotic leaf spot virus (ACLSV), stem grooving (ASGV), and pitting (ASPV) are three latent viruses frequently occurring in apple trees worldwide. In field orchards, these found a mixed infection with viroids the genus Apscarviroid, including scar skin viroid, dimple fruit viroid. Together could cause serious damage to production Rapid efficient detection methods pivotal identify select virus-free propagation material for healthy orchard management. this study multiplex Reverse Transcription-PCR (RT-PCR) was developed optimized simultaneous differentiation of apscarviroids. With newly designed specific primers ACLSV, ASGV, APSV, EF-1α (as an internal control), pair degenerate apscarviroids, parameters RT-PCR were determined. The resulting PCR products from each target viroid be easily identified because their product sizes differ by at least 100bp. method is expected detect different variants as test results showed that variety isolates regions China gave positive results. To best our knowledge, assay first simultaneously multiple infecting single reaction tube. This assay, therefore, offers useful tool routine certification quarantine programs.