Journal
LETTERS IN APPLIED MICROBIOLOGY
Volume 62, Issue 5, Pages 386-391Publisher
WILEY-BLACKWELL
DOI: 10.1111/lam.12569
Keywords
African swine fever virus; cross-priming amplification; development; direct virus DNA detection; pigs and wild boar blood; sera
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Funding
- KNOW (Leading National Research Centre) Scientific Consortium 'Healthy Animal - Safe Food', Ministry of Science and Higher Education [05-1/KNOW2/2015 (K/02/1.0)]
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African swine fever (ASF) is considered a major threat to the production of pigs worldwide. The ASF aetiological agent, ASFV, is the sole member of the Asfivirus genus, belonging to the Asfarviridae family. An effective ASF vaccine is not currently available, thus the only measures of ASF spread control include, reliable and fast diagnosis. Officially approved, diagnostic methods include, virus isolation, serological assays, including enzyme-linked immunosorbent assay and immunoperoxidase assay (IPT) and different modifications of the polymerase chain reaction (PCR). This paper describes the first development and application of a cross-priming amplification method (CPA) for the direct detection of genetic ASFV material, in blood and sera from pigs and wild boars. This method is specific only to ASFV DNA. The study showed that CPA had equal sensitivity, in comparison to the official, universal probe library (UPL) real-time PCR and reached 7.2 copies of standard plasmid DNA, containing a p72 gene fragment. This method was capable of detecting ASFV DNA in all examined blood samples, originating from pigs; n=10 and wild boars; n=76. The obtained results were also confirmed by the officially approved, real-time PCR. The developed CPA might be further used by local and county veterinary officers, hunters or pig farmers, for preliminary ASF diagnosis.
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