Abstract
In the recent years, African swine fever has become the biggest animal health threat to the swine industry. To facilitate quick genetic analysis of its causative agent, the African swine fever virus (ASFV), we developed a simple and efficient method for next generation sequencing of the viral DNA. Execution of the protocol does not demand complicated virus purification steps, enrichment of the virus by ultracentrifugation or of the viral DNA by ASFV-specific PCRs, and minimizes the use of Sanger sequencing. Efficient DNA-se treatment, monitoring of sample preparation by qPCR, and whole genome amplification are the key elements of the method. Through detailed description of sequencing of the first Hungarian ASFV isolate (ASFV_HU_2018), we specify the sensitive steps and supply key reference numbers to assist reproducibility and to facilitate the successful use of the method for other ASFV researchers.
Highlights
African swine fever virus (ASF) is a devastating disease affecting Sus scrofa; it infects both domesticated pigs and wild boars
The virus was introduced into Europe in several successive waves; genotype I virus spread to the Southern Europe in the 1950’s and 1960’s, while the well documented emergence of genotype II African swine fever virus (ASFV) occurred in Georgia in 2007 [6]
Similar problems were reported at the sequencing of other ASFV isolates and different methodical approaches were pursued to overcome the difficulties [28,29,30,31,32]
Summary
African swine fever virus (ASF) is a devastating disease affecting Sus scrofa; it infects both domesticated pigs and wild boars. The ASF virus (ASFV) was most probably transmitted from its natural hosts, warthogs (Phacochoerus spp.), bushpigs (Potamochoerus spp.) and soft ticks of the genus. Ornithodoros to domestic pigs (Sus scrofa domesticus) in southeast Africa [1,2]. The virus is endemic in the Sub-Saharan region, where viral reservoir is maintained by a sylvatic cycle between the soft ticks and its natural hosts [3,4]. The virus was introduced into Europe in several successive waves; genotype I virus spread to the Southern Europe in the 1950’s and 1960’s, while the well documented emergence of genotype II ASFV occurred in Georgia in 2007 [6]. Since 2017 the virus has continuously spread westward in Eastern Europe, reaching
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