PCR assay to detect Xylophilus ampelinus in grapevine cuttings
A PCR assay has been developed in South Africa to detect specifically Xylophilus ampelinus (EPPO A2 quarantine pest) in grapevine cuttings. 16S-23S rDNA intergenic spacer region of X. ampelinus was sequenced and specific primers were designed to target a part of this sequence. A nested-PCR assay was then developed using these primers for routine detection of X. ampelinus in the sap of grapevine cuttings. The authors noted that their method can detect low numbers of bacteria, even in the presence of saprophytic bacteria commonly associated with grapevine sap (e.g. less than 10 CFU µL-1 even in the presence of 1.5 x105 CFU µL-1 of Erwinia herbicola cells). They concluded that their method is quick and reliable in comparison with the time-consuming and unreliable conventional method of plating samples on growing media followed by purification of bacterial colonies.
Botha, W.J.; Serfontein, S.; Greyling, M.M.; Berger, D.K. (2001) Detection of Xylophilus ampelinus in grapevine cuttings using a nested polymerase chain reaction.
Plant Pathology, 50(4), 515-526.