Abstract
Molecular biology methods are highly sensitive to detect the genome of pathogens and to study their biology. Polymerase chain reaction (PCR) and reverse transcription followed by a polymerase chain reaction (RT-PCR) permit the detection of the presence and the replication of African swine fever virus in soft ticks. Here, we described our techniques to detect and quantify DNA and RNA of African swine fever virus in soft ticks including a housekeeping gene of soft ticks as internal control.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ravaomanana J, Michaud V, Jori F et al (2010) First detection of African swine fever virus in ornithodoros porcinus in Madagascar and new insights into tick distribution and taxonomy. Parasites Vectors 3:115. https://doi.org/10.1186/1756-3305-3-115
Basto AP (2006) Kinetics of African swine fever virus infection in Ornithodoros erraticus ticks. J Gen Virol 87:1863–1871. https://doi.org/10.1099/vir.0.81765-0
Basto AP, Portugal RS, Nix RJ et al (2006) Development of a nested PCR and its internal control for the detection of African swine fever virus (ASFV) in Ornithodoros erraticus. Arch Virol 1501:819–826. https://doi.org/10.1007/s00705-005-0654-2
Chastagner A, Pereira de Oliveira R, Hutet E et al (2020) Coding-complete genome sequence of an African Swine Fever Virus strain Liv13/33 isolate from experimental transmission between pigs and ornithodoros moubata ticks. Microbiol Resour Announc 9, e00185–20, /mra/9/17/MRA.00185-20.atom. https://doi.org/10.1128/MRA.00185-20
de Carvalho Ferreira HC, Tudela Zúquete S, Wijnveld M et al (2014) No evidence of African swine fever virus replication in hard ticks. Ticks and Tick-borne Diseases 5:582–589. https://doi.org/10.1016/j.ttbdis.2013.12.012
Pereira De Oliveira R, Hutet E, Duhayon M et al (2020) Successful infection of domestic pigs by ingestion of the European soft tick O. Erraticus that fed on African swine fever virus infected pig. Viruses 12:300. https://doi.org/10.3390/v12030300
Pereira De Oliveira R, Hutet E, Duhayon M et al (2020) No experimental evidence of co-feeding transmission of African swine fever virus between ornithodoros soft ticks. Pathogens 9:168. https://doi.org/10.3390/pathogens9030168
Pereira De Oliveira R, Hute E, Lancelot R et al (2020) Differential vector competence of Ornithodoros soft ticks for African swine fever virus: what if it involves more than just crossing organic barriers in ticks? Parasites Vectors 13:618. https://doi.org/10.1186/s13071-020-04497-1
Pereira de Oliveira R, Hutet E, Paboeuf F et al (2019) Comparative vector competence of the Afrotropical soft tick Ornithodoros moubata and Palearctic species, O. erraticus and O. verrucosus, for African swine fever virus strains circulating in Eurasia. PLoS ONE 14:e0225657. https://doi.org/10.1371/journal.pone.0225657
Pereira De Oliveira R, Lucas P, Chastagner A et al (2020) Evaluation of un-methylated DNA enrichment in sequencing of African swine fever virus complete genome. J Virol Met 285:113959. https://doi.org/10.1016/j.jviromet.2020.113959
Forth JH, Forth LF, Lycett S et al (2020) Identification of African swine fever virus-like elements in the soft tick genome provides insights into the virus’ evolution. BMC Biol 18:136. https://doi.org/10.1186/s12915-020-00865-6
Tignon M, Gallardo C, Iscaro C et al (2011) Development and inter-laboratory validation study of an improved new real-time PCR assay with internal control for detection and laboratory diagnosis of African swine fever virus. J Virol Methods 178:161–170. https://doi.org/10.1016/j.jviromet.2011.09.007
Duron O, Morel O, Noël V et al (2018) Tick-bacteria mutualism depends on B vitamin synthesis pathways. Curr Biol 28:1896–1902.e5. https://doi.org/10.1016/j.cub.2018.04.038
Fernández-Pinero J, Gallardo C, Elizalde M et al (2013) Molecular diagnosis of African swine fever by a new real-time PCR using universal probe library: UPL PCR for African swine fever virus. Transbound Emerg Dis 60:48–58. https://doi.org/10.1111/j.1865-1682.2012.01317.x
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Pereira De Oliveira, R., Vial, L., Le Potier, MF. (2022). Quantification of ASFV DNA and RNA in Ornithodoros Soft Ticks. In: Netherton, C.L. (eds) African Swine Fever Virus. Methods in Molecular Biology, vol 2503. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2333-6_7
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2333-6_7
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2332-9
Online ISBN: 978-1-0716-2333-6
eBook Packages: Springer Protocols