Journal article
Specific detection of the Old World screwworm fly, Chrysomya bezziana, in bulk fly trap catches using real-time PCR
Medical and Veterinary Entomology, Vol.24(3), pp.227-235
2010
Abstract
The Old World screwworm fly (OWS), Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), is a myiasis-causing blowfly of major concern for both animals and humans. Surveillance traps are used in several countries for early detection of incursions and to monitor control strategies. Examination of surveillance trap catches is time-consuming and is complicated by the presence of morphologically similar flies that are difficult to differentiate from Ch. bezziana, especially when the condition of specimens is poor. A molecular-based method to confirm or refute the presence of Ch. bezziana in trap catches would greatly simplify monitoring programmes. A species-specific real-time polymerase chain reaction (PCR) assay was designed to target the ribosomal DNA internal transcribed spacer 1 (rDNA ITS1) of Ch. bezziana. The assay uses both species-specific primers and an OWS-specific Taqman® MGB probe. Specificity was confirmed against morphologically similar and related Chrysomya and Cochliomyia species. An optimal extraction protocol was developed to process trap catches of up to 1000 flies and the assay is sensitive enough to detect one Ch. bezziana in a sample of 1000 non-target species. Blind testing of 29 trap catches from Australia and Malaysia detected Ch. bezziana with 100% accuracy. The probability of detecting OWS in a trap catch of 50 000 flies when the OWS population prevalence is low (one in 1000 flies) is 63.6% for one extraction. For three extractions (3000 flies), the probability of detection increases to 95.5%. The real-time PCR assay, used in conjunction with morphology, will greatly increase screening capabilities in surveillance areas where OWS prevalence is low.
Details
- Title
- Specific detection of the Old World screwworm fly, Chrysomya bezziana, in bulk fly trap catches using real-time PCR
- Authors/Creators
- S. Jarrett (Author/Creator) - Department of Jobs and Small BusinessJ.A.T. Morgan (Author/Creator) - Department of Jobs and Small BusinessB.M. Wlodek (Author/Creator) - Department of Jobs and Small BusinessG.W. Brown (Author/Creator) - Department of Jobs and Small BusinessR. Urech (Author/Creator) - Department of Jobs and Small BusinessP.E. Green (Author/Creator) - Department of Jobs and Small BusinessA.E. Lew-Tabor (Author/Creator) - Department of Jobs and Small Business
- Publication Details
- Medical and Veterinary Entomology, Vol.24(3), pp.227-235
- Publisher
- Blackwell Publishing Inc.
- Identifiers
- 991005543585307891
- Copyright
- © 2010 State of Queensland, Department of Employment, Economic Development and Innovation
- Murdoch Affiliation
- Centre for Comparative Genomics
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 3 Agriculture, Environment & Ecology
- 3.32 Entomology
- 3.32.1638 Forensic Entomology
- Web Of Science research areas
- Entomology
- Veterinary Sciences
- ESI research areas
- Plant & Animal Science