A simplified RT-PCR-based detection of recombinant Plum pox virus isolates.
Closely related natural Plum pox virus (PPV) isolates derived from homologous RNA recombination between PPV-D and PPV-M have been recently identified and shown naturally spread in several European countries. As their serological properties were identical with those of conventional PPV-M isolates, they could be detected only by combined analysis of at least two different genome parts. To simplify the detection of such recombinants primers specific to PPV-M and PPV-D sequences with binding sites located on both sides of the recombination crossover situated in the C-terminal part of NIb were designed. They were used for direct differentiation of PPV-M, PPV-D and their recombinants by reverse transcription-polymerase chain reaction (RT-PCR). This method is convenient for identification of a recombinant PPV in single as well as mixed infection with PPV-M or PPV-D.
Subr
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Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 845 05 Bratislava, Slovak Republic. virusubr@savba.sk
Pittnerová
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Glasa
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D017931:DNA Primers; D017801:Plum Pox Virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D027861:Prunus; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012703:Serotyping / Q000379:methods; D013045:Species Specificity; D020079:Terminal Repeat Sequences; D014764:Viral Proteins / Q000235:genetics
D017931:DNA Primers; D014764:Viral Proteins