Rapid Identification for Serotyping of African Swine Fever Virus Based on the Short Fragment of the EP402R Gene Encoding for CD2-Like Protein

Abstract

The first confirmed case of African swine fever (ASF) in Vietnam was officially reported in February 2019. To date, the ASF virus (ASFV) has been detected in all 63 cities/provinces in Vietnam. In order to get a better understanding of the potential role of the EP402R gene in a grouping of ASFV serotypes, thirty ASFV sequences of EP402R genes (accession numbers: MN711757-86) from North Central Coast of Vietnam and 68 well-known references of serotype groups from previous studies were further analyzed. Interestingly, we found that a short fragment of 90 nucleotides was very typical for 8 serological groups of ASFVs. A primer set was designed to amplicon the short fragment of 90 nucleotides using the Primer3 program to establish a simplified method for the serotyping of ASFV. Our results indicated that phylogenetic analysis of the short fragment (90 nucleotides) of the EP402R gene is a very specific and useful method for ASFV serotyping when compared to the previous method using a long fragment (816 nucleotides) of this gene and well-known serotype references based on haemadsorption inhibition (HAI) assay.

Prvi potvrđen slučaj afričke kuge svinja (ASF) u Vijetnamu bio je zvanično prijavljen u februaru 2019. godine. Do danas, ASF virus (ASFV) je detektovan u 63 provincije Vijetnama. Radi boljeg razumevanja potencijalne uloge EP402R gena u grupisanju ASFV serotipova, analizirano je trideset ASFV sekvenci EP402R gena (pristupni brojevi: MN711757-86) poreklom iz Centralne severne obale Vijetnama i 68 referentnih uzoraka serotip grupa koji su dobro poznati iz prethodnih studija. Interesantan je bio nalaz da je kratak fragment od 90 nukleotida bio tipičan za 8 seroloških grupa ASF virusa. Dizajniran je set prajmera sa ciljem amplifi kacije kratkih fragmenata (90 nukleotida), korišćenjem Primer3 programa, a za uspostavljanje jednostavne metode serotipiziranja ASFV. Rezultati ukazuju da je fi logenetska analiza kratkih fragmenata (90 nukleotida) EP402R gena je veoma specifična i predstavlja korisnu metodu serotipizacije ASFV u poređenju sa prethodno opisanim metodama koje su koristile duge fragmente (816 nukleotida) ovog gena kao i dobro poznate referentne serotipove dobijene metodom inhibicije hemadsorbcije.