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Detection of Xylella fastidiosa in potential insect vectors by immunomagnetic separation and nested polymerase chain reaction
Author(s) -
Pooler M.R.,
Myung I.S.,
Bentz J.,
Sherald J.,
Hartung J.S.
Publication year - 1997
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.1997.00188.x
Subject(s) - xylella fastidiosa , biology , nested polymerase chain reaction , leafhopper , polymerase chain reaction , bacteria , vector (molecular biology) , insect , microbiology and biotechnology , virology , botany , genetics , gene , recombinant dna , hemiptera
A sensitive and specific assay for detecting Xylella fastidiosa in potential insect vectors was developed. This assay involves immunomagnetic separation of the bacteria from the insect, followed by a two‐step, nested polymerase chain reaction (PCR) amplification using previously developed oligonucleotide primers specific to X. fastidiosa . A total of 347 leafhoppers representing 16 species were captured and sampled from American elm ( Ulmus americana L.) trees growing in a nursery where bacterial leaf scorch caused by X. fastidiosa occurs. Two of these leafhopper species, Graphocephala coccinea and G. versuta , regularly tested positive for X. fastidiosa using this technique. These insects are therefore potential vectors of X. fastidiosa . Using immunocapture and nested PCR, it was possible to detect as few as five bacteria per sample.