Diagnosis of Lobesia botrana (Lepidoptera: Tortricidae) Using Real-Time PCR

A real-time PCR assay is reported for identification of Lobesia botrana (Denis and Schiffermüller) collected in California. This assay multiplexes two independent TaqMan probe systems in a single reaction tube to reduce handling time and sample exposure to environmental contaminants. One probe syste...

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Bibliographic Details
Published in:Journal of economic entomology 2016-08, Vol.109 (4), p.1957-1962
Main Authors: Ledezma, L. A, Barr, N. B, Epstein, M. E, Gilligan, T. M
Format: Article
Language:English
Subjects:
Animal populations
Animals
Butterflies & moths
California
Contaminants
Deoxyribonucleic acid
DNA
DNA, Ribosomal Spacer - analysis
European grape vine moth
Insect Control - methods
internal transcribed spacer
Larva - classification
Larva - genetics
Moths - classification
Moths - genetics
Paralobesia
Polymerase chain reaction
Population genetics
Pupa - classification
Pupa - genetics
Real-Time Polymerase Chain Reaction - methods
RNA, Ribosomal, 18S - analysis
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Summary:A real-time PCR assay is reported for identification of Lobesia botrana (Denis and Schiffermüller) collected in California. This assay multiplexes two independent TaqMan probe systems in a single reaction tube to reduce handling time and sample exposure to environmental contaminants. One probe system targets a segment of DNA located in the internal transcribed spacer region 2 (ITS2) that is present in the L. botrana genome but absent in native North American Tortricidae. The second probe system serves as a control for DNA quality by targeting a segment of the 18S rDNA gene that is conserved in L. botrana and all of the tested nontarget species. The assay successfully diagnosed 70 Lobesia botrana specimens and 95 nontarget specimens. No false-positive or false-negative results were observed supporting its application for identification of this pest in California.
ISSN:0022-0493
1938-291X
DOI:10.1093/jee/tow111