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Insects 2015, 6(3), 746-759;

Development of a Real-Time qPCR Assay for Quantification of Covert Baculovirus Infections in a Major African Crop Pest

Lancaster Environment Centre, Lancaster University, Lancaster LA1 4YQ, UK
Lake Ecosystems Group, Centre for Ecology and Hydrology, Bailrigg, Lancaster LA1 4AP, UK
Department of Biological Sciences, Simon Fraser University, Burnaby, BC V5A 1S6, Canada
Biomedical and Life Sciences, Lancaster University, Lancaster LA1 4YQ, UK
Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent ME4 4TB, UK
Current address: Crop and Environment Sciences, Harper Adams University, Edgmond, Shropshire TF10 8NB, UK.
Author to whom correspondence should be addressed.
Academic Editor: Brian T. Forschler
Received: 23 April 2015 / Revised: 14 August 2015 / Accepted: 19 August 2015 / Published: 25 August 2015
(This article belongs to the Special Issue Parasite-Insect Interactions)
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Many pathogens and parasites are present in host individuals and populations without any obvious signs of disease. This is particularly true for baculoviruses infecting lepidopteran hosts, where studies have shown that covert persistent viral infections are almost ubiquitous in many species. To date, the infection intensity of covert viruses has rarely been quantified. In this study, we investigated the dynamics of a covert baculovirus infection within the lepidopteran crop pest Spodoptera exempta. A real-time quantitative polymerase chain reaction (qPCR) procedure using a 5' nuclease hydrolysis (TaqMan) probe was developed for specific detection and quantification of Spodoptera exempta nucleopolyhedrovirus (SpexNPV). The qPCR assay indicated that covert baculovirus dynamics varied considerably over the course of the host life-cycle, with infection load peaking in early larval instars and being lowest in adults and final-instar larvae. Adult dissections indicated that, contrary to expectation, viral load aggregation was highest in the head, wings and legs, and lowest in the thorax and abdomen. The data presented here have broad implications relating to our understanding of transmission patterns of baculoviruses and the role of covert infections in host-pathogen dynamics. View Full-Text
Keywords: Spodoptera exempta; baculovirus; nucleopolyhedrovirus; covert infections; TaqMan real-time qPCR Spodoptera exempta; baculovirus; nucleopolyhedrovirus; covert infections; TaqMan real-time qPCR

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Graham, R.I.; Tummala, Y.; Rhodes, G.; Cory, J.S.; Shirras, A.; Grzywacz, D.; Wilson, K. Development of a Real-Time qPCR Assay for Quantification of Covert Baculovirus Infections in a Major African Crop Pest. Insects 2015, 6, 746-759.

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