A host species-informative internal control for molecular assessment of African swine fever virus infection rates in the African sylvatic cycle Ornithodoros vector

Medical and Veterinary Entomology
A D S BastosS Maree

Abstract

African swine fever virus (ASFV) infection in adult Ornithodoros porcinus (Murry 1877, sensuWalton 1979) ticks collected from warthog burrows in southern and East Africa was assessed using a duplex genomic amplification approach that is informative with respect to the invertebrate host species and infecting sylvatic cycle virus. DNA extracted from individual ticks was used as template for the simultaneous amplification of a C-terminal 478-bp ASFV p72 gene region and a approximately 313-bp fragment of the tick mitochondrial 16S rRNA gene, under optimized reaction conditions. Within-warthog burrow infection rates ranged from 0% to 43% using this approach, and phylogenetic analysis of 16S gene sequences revealed the presence of three geographically discrete O. porcinus lineages, but no support for subspecies recognition. False negatives are precluded by the inclusion of host species-informative primers that ensure the DNA integrity of cytoplasmically located genome extracts. In addition, infection rate estimates are further improved as false positives arising from carry-over contamination when performing a two-step nested polymerase chain reaction are negated by the one-step approach. Phylogenetic comparison of full-length virus g...Continue Reading

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Citations

Jul 2, 2014·Ticks and Tick-borne Diseases·Helena C de Carvalho FerreiraWillie L A Loeffen
Nov 13, 2012·Virus Research·Mary-Louise PenrithArmanda D S Bastos
Nov 6, 2012·Virus Research·S CostardD U Pfeiffer
May 24, 2016·Transboundary and Emerging Diseases·J E AchenbachJ M Sánchez-Vizcaíno
Jun 2, 2016·The Onderstepoort Journal of Veterinary Research·Noluvuyo R MagadlaBruce Gummow
Jan 1, 2014·The Onderstepoort Journal of Veterinary Research·Carin I BoshoffLivio Heath
Nov 17, 2009·EcoHealth·Ferran Jori, Armanda D S Bastos
Jul 23, 2020·Transboundary and Emerging Diseases·Emma PeterRoger Pelle
Jun 2, 2020·Frontiers in Veterinary Science·Natasha N GaudreaultJuergen A Richt

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