Detection of Xylella fastidiosa in potential insect vectors by immunomagnetic separation and nested polymerase chain reaction |
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| Authors: | M.R. Pooler,I.S. Myung,J. Bentz,J. Sherald,& J.S. Hartung |
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| Affiliation: | US National Arboretum, United States Department of Agriculture, Washington, DC,;Division of Overseas Pests, Department of Crop Protection, National Institute of Agricultural Science and Technology, Suwon, Korea,;Floral and Nursery Plants Research Unit,;Fruit Laboratory, Agricultural Research Service, US Department of Agriculture, Beltsville Agricultural Research Center, Beltsville, MD,;USDI, National Park Service, National Capital Area, Center for Urban Ecology, Washington, DC, USA |
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| Abstract: | 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. |
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