Single-Step Multiplex Reverse Transcription-Polymerase Chain Reaction (m-RT-PCR) for Simultaneous Detection of Five RNA Viruses Affecting Stone Fruit Trees
Single-Step Multiplex Reverse Transcription-Polymerase Chain Reaction (m-RT-PCR) for Simultaneous Detection of Five RNA Viruses Affecting Stone Fruit Trees
Sahar A. Youssef I and A. A. Shalaby
ABSTRACT
A single-step multiplex reverse transcription-polymerase chain reaction (m-RTPCR) was developed for the simultaneous detection and discrimination among five RNA viruses namely ,4pple cholorotic leaf spot virus (ACLSV), Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV), Plum pox virus (PPV) and Tomato ringspot virus
(ToRSV) which considered the most economically damaging viruses of stone fruit trees in Egypt and worldwide. Five compatible primer sets for one•step RT-PCR amplification used in a single closed tube to detect the presence of virus infection. Five fragments with different sizes corresponding to the different viral targets were amplified from infected samples, .i.e., 677, 172, 300, 270 and bp for ACLSV, PDV, PNRSV, PPV and ToRSV respectively, Results showed that multiplex PCR products of the expected sizes were obtained for all virus isolates assayed they were tested either alone or in c.ombination in an artificial mixture with no interference. This method saves time and reagent costs compared with rnonospecific RT-PCR needs several reactions for the sarne number of tests. Using this method, jnany samples can be simultaneously analyzed and it can be used routinely för sanitary and cettification prograrnmes.
To share on other social networks, click on any share button. What are these?
July 2008
Vol. 5, Iss. 1
