Development of cross-priming amplification for direct detection of the African Swine Fever Virus, in pig and wild boar blood and sera samples
| Autor: | Grzegorz Woźniakowski, Andrzej Kowalczyk, K. Niemczuk, Magdalena Frączyk, Z. Pejsak |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Swine Molecular Sequence Data Sus scrofa Enzyme-Linked Immunosorbent Assay Biology Real-Time Polymerase Chain Reaction Applied Microbiology and Biotechnology African swine fever virus law.invention Serology 03 medical and health sciences chemistry.chemical_compound Wild boar law Asfarviridae biology.animal Animals African Swine Fever Gene Polymerase chain reaction Immunoperoxidase biology.organism_classification Virology African Swine Fever Virus 030104 developmental biology chemistry DNA Viral Nucleic Acid Amplification Techniques DNA |
| Zdroj: | Letters in applied microbiology. 62(5) |
| ISSN: | 1472-765X |
| Popis: | UNLABELLED 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. SIGNIFICANCE AND IMPACT OF THE STUDY The spread of the African swine fever virus (ASFV) among infected pigs and wild boars, is currently one of the most important facets of virus transmission in eastern Europe. Cross-priming amplification (CPA) has been developed, for fast and direct development of genetic ASFV material in the blood and sera of infected pigs and wild boars. It has been shown that CPA is a rapid, sensitive and specific isothermal method for the detection of ASFV DNA, in directly collected blood or sera from pigs and wild boars. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text