Fast, easy and efficient DNA extraction and one-step polymerase chain reaction for the detection of Xylella fastidiosa in potential insect vectors
A quick, simple and efficient procedure for detecting Xylella fastidiosa in potential insect vectors is described. The procedure employs a commercially available DNeasy tissue kit for the extraction of high-quality DNA from the insect, followed by one-step polymerase chain reaction amplification using previously published oligonucleotide primers specific to X fastidiosa. The procedure does not require the use of phenol, chloroform or alcohol for the precipitation of nucleic acids. Also it does not need additional purification or enrichment steps, and can be completed in less than a day. The procedure was used successfully in detecting X. fastidiosa in two potentially important leafhopper species, Graphocephala versuta and G. coccinea, and in a treehopper species Entilia concisa, collected from a nursery where bacterial leaf scorch disease caused by X. fastidiosa occurs.