Multiplex RT-PCR Assay for Simultaneous Detection of Three Latent Apple Viruses
FAN Xu-dong1;DONG Ya-feng1*;ZHANG Zun-ping1 ;LI Li-li2 ; PEI Guang-qian
(1Research Institute of Pomology, Chinese Academy of Agriculture Sciences, Xingcheng, Liaoning 125100, China; 2College of Horticulture, Shenyang Agricultural University, Shenyang 110161, China)
Abstract：The multiplex RT-PCR assay was established to simultaneously detect Apple stem grooving virus (ASGV) , Apple chlorotic leaf spot virus (ACLSV) and Apple stem pitting virus (ASPV). Using tissue culture apple seedling (Wangshanhong) infected by three latent apple viruses as sample, a series of factors affecting the multip lex RT-PCR were optimized. Sensibility test showed that it could simultaneously detect out ASGV, ACLSV and ASPV in 187.5 ng total RNA. The multiplex RT-PCR products were cloned and sent for sequencing, and the sequences show high homologies to the published virus sequences. Consistent results for virus detection of 12 apple samples in field were obtained by the multiplex RT-PCR and single RT-PCR system, suggesting the accuracy of the system in ASGV, ACLSV and ASPV detection.
2009, Vol. 36 
