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. 2002 Dec;106(2):235-9.
doi: 10.1016/s0166-0934(02)00147-7.

Simultaneous Detection of Six Citrus Viroids and Apple Stem Grooving Virus From Citrus Plants by Multiplex Reverse Transcription Polymerase Chain Reaction

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Simultaneous Detection of Six Citrus Viroids and Apple Stem Grooving Virus From Citrus Plants by Multiplex Reverse Transcription Polymerase Chain Reaction

Takao Ito et al. J Virol Methods. .

Abstract

We developed a multiplex reverse transcription polymerase chain reaction (RT-PCR) to detect six citrus viroids: Citrus exocortis viroid (CEVd), Citrus bent leaf viroid (CBLVd), Hop stunt viroid (HSVd), Citrus viroid III (CVd-III), Citrus viroid IV (CVd-IV) and Citrus viroid OS (CVd-OS) and Apple stem grooving virus (ASGV, synonym: Citrus tatter leaf virus (CTLV)) from citrus plants. The multiplex RT-PCR was also designed to distinguish CVd-I-LSS (a distinct variant of CBLVd) from CBLVd. By the multiplex RT-PCR, one to eight fragments specific to the pathogens were simultaneously amplified from one sample and identified by their specific molecular sizes in 6% polyacrylamide gel electrophoresis. The results of the multiplex RT-PCR were consistent with those of other diagnoses, such as uniplex RT-PCR, to detect each of the pathogens. The multiplex RT-PCR provides a simple and rapid method for detecting various viroids and ASGV in citrus plants, which will help diagnose many citrus plants at a time.

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