Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay

Simple item page
dc.contributor.authorMurray, Lee
dc.contributor.authorEdwards, Lorraine
dc.contributor.authorTuppurainen, Eeva SM
dc.contributor.authorBachanek-Bankowska, Katarzyna
dc.contributor.authorOura, Chris AL
dc.contributor.authorMioulet, Valerie
dc.contributor.authorKing, Donald P
dc.date.accessioned2014-04-11T13:30:42Z
dc.date.available2014-04-11T13:30:42Z
dc.date.issued2013-05-01
dc.date.updated2014-04-11T13:30:42Z
dc.description.abstractAbstract Background Sheep poxvirus (SPPV), Goat poxvirus (GTPV) and Lumpy skin disease virus (LSDV) are the most serious poxviruses of ruminants. They are double stranded DNA viruses of the genus Capripoxvirus, (subfamily Chordopoxvirinae) within the family Poxviridae. The aim of this study was to develop a Loop-mediated isothermal AMPlification (LAMP) assay for the detection of Capripoxvirus (CaPV) DNA. Results A single LAMP assay targeting a conserved region of the CaPV P32 gene was selected from 3 pilot LAMP assays and optimised by adding loop primers to accelerate the reaction time. This LAMP assay successfully detected DNA prepared from representative CaPV isolates (SPPV, GTPV and LSDV), and did not cross-react with DNA extracted from other mammalian poxviruses. The analytical sensitivity of the LAMP assay was determined to be at least 163 DNA copies/and#956;l which is equivalent to the performance reported for diagnostic real-time PCR currently used for the detection of CaPV. LAMP reactions were monitored with an intercalating dye using a real-time PCR machine, or by agarose-gel electrophoresis. Furthermore, dual labelled LAMP products (generated using internal LAMP primers that were conjugated with either biotin or fluorescein) could be readily visualised using a lateral-flow device. Conclusions This study provides a simple and rapid approach to detect CaPV DNA that may have utility for use in the field, or in non-specialised laboratories where expensive equipment is not available.
dc.description.versionPeer Reviewed
dc.identifier.citationBMC Veterinary Research. 2013 May 01;9(1):90
dc.identifier.urihttp://dx.doi.org/10.1186/1746-6148-9-90
dc.identifier.urihttps://hdl.handle.net/2139/38061
dc.language.rfc3066en
dc.rights.holderLee Murray et al.; licensee BioMed Central Ltd.
dc.titleDetection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
dc.typeJournal Article

Files

Original bundle
Now showing 1 - 2 of 2
No Thumbnail Available
Name:
1746-6148-9-90.xml
Size:
76.41 KB
Format:
Extensible Markup Language
Download
Thumbnail Image
Name:
1746-6148-9-90.pdf
Size:
860.75 KB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

BioMed Central