Multiplex RT-PCR Assay for Simultaneous Detection of Three Latent Apple Viruses

ACTA HORTICULTURAE SINICA ›› 2009, Vol. 36 ›› Issue (12): 1821-1826.

• Technologies and Methods • Previous Articles Next Articles

Multiplex RT-PCR Assay for Simultaneous Detection of Three Latent Apple Viruses

FAN Xu-dong¹;DONG Ya-feng^1*;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)

Received:2009-07-22 Revised:2009-10-30 Online:2009-12-25 Published:2009-12-25
Contact: DONG Ya-feng

Abstract

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.

Key words: apple, multiplex RT-PCR, detection, Apple stem grooving virus, Apple chlorotic leaf spotvirus, Apple stem pitting virus

CLC Number:

S661.1

FAN Xu-dong;DONG Ya-feng;ZHANG Zun-ping;LI Li-li;PEI Guang-qian. Multiplex RT-PCR Assay for Simultaneous Detection of Three Latent Apple Viruses[J]. ACTA HORTICULTURAE SINICA, 2009, 36(12): 1821-1826.

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Multiplex RT-PCR Assay for Simultaneous Detection of Three Latent Apple Viruses

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