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An improved real-time PCR assay for the detection of Old World screwworm flies

Morgan, J. A. T. and Urech, R. (2014) An improved real-time PCR assay for the detection of Old World screwworm flies. Acta Tropica . ISSN 0001706X

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Article Link(s): http://dx.doi.org/10.1016/j.actatropica.2014.02.01...

Abstract

The Old World screwworm (OWS) fly, Chrysomya bezziana, is a serious pest of livestock, wildlife and humans in tropical Africa, parts of the Middle East, the Indian subcontinent, south-east Asia and Papua New Guinea. Although to date Australia remains free of OWS flies, an incursion would have serious economic and animal welfare implications. For these reasons Australia has an OWS fly preparedness plan including OWS fly surveillance with fly traps. The recent development of an improved OWS fly trap and synthetic attractant and a specific and sensitive real-time PCR molecular assay for the detection of OWS flies in trap catches has improved Australia's OWS fly surveillance capabilities. Because all Australian trap samples gave negative results in the PCR assay, it was deemed necessary to include a positive control mechanism to ensure that fly DNA was being successfully extracted and amplified and to guard against false negative results. A new non-competitive internal amplification control (IAC) has been developed that can be used in conjunction with the OWS fly PCR assay in a multiplexed single-tube reaction. The multiplexed assay provides an indicator of the performance of DNA extraction and amplification without greatly increasing labour or reagent costs. The fly IAC targets a region of the ribosomal 16S mitochondrial DNA which is conserved across at least six genera of commonly trapped flies. Compared to the OWS fly assay alone, the multiplexed OWS fly and fly IAC assay displayed no loss in sensitivity or specificity for OWS fly detection. The multiplexed OWS fly and fly IAC assay provides greater confidence for trap catch samples returning negative OWS fly results. © 2014 International Atomic Energy Agency.

Item Type:Article
Business groups:Animal Science
Additional Information:Correspondence Address: Morgan, J.A.T.; Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, Queemail: Jessica.Morgan@uq.edu.au
Keywords:Chrysomya bezziana Diagnostics IAC mtDNA Multiplex Myiasis
Subjects:Science > Invasive Species > Animals
Veterinary medicine > Veterinary parasitology
Science > Entomology
Deposited On:19 Aug 2014 02:55
Last Modified:19 Aug 2014 02:55

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