LM preformed sequence alignment and analysis of the pyrosequencing data. LB participated in the design of the molecular study. JDV and FVI designed and conducted the experimental studies,
and KP conceived of the study. All authors read and approved the final manuscript. The authors declare that they have no competing interests.”
“Background Porcine circovirus (PCV) is the smallest #buy SB202190 randurls[1|1|,|CHEM1|]# virus that replicates autonomously in mammalian cells. The viral genome consists of a covalently closed, circular, ambisense, single-stranded DNA molecule [1]. Two types of PCV (1 and 2), have been characterized to date [2]. PCV1 is a persistent contaminant of porcine kidney (PK)-15 cell lines and it is not considered to be pathogenic [3]. In contrast, PCV2 has been detected consistently in pigs with PCV-associated diseases such as post-weaning multisystemic wasting syndrome (PMWS) [4]. The genome of PCV2 contains at least two open reading frames (ORFs) with known functions: ORF1 codes for two replicase proteins, and ORF2 for the structural capsid protein [5]. The capsid protein is the only structural protein and the major protein involved in immunogenicity. At least five overlapping conformational epitopes of PCV2 capsid protein, within residues 47-85, 165-200 and 230-233, have been
mapped in chimeric PCV1 and PCV2 [6]. The conformational epitopes recognized by monoclonal antibodies (mAbs) with neutralizing activity against
PCV2 click here have been determined in the transfected PK-15 cells, and residues 231-233 participate in the formation of conformational epitopes [7]. Phylogenetic analysis distinguishes three genotypes of Chorioepithelioma PCV2 (a, b and c) [8]. PCV2a and PCV2b are found in many countries, whereas PCV2c is only found in Denmark [9]. Recent epidemiological studies in many countries have linked a shift from infection with PCV2a to PCV2b [9–12]. Although several studies have indicated that PCV2b is not more pathogenic than PCV2a [13], field experience suggests that the PCV2b genotype is more virulent [11]. However, to date, there are no confirmed conclusions about which genotype is more pathogenic. Mouse mAbs directed against PCV2 have shown some differences in reactivity with different PCV2 strains [7, 14]. MAbs (with different reactivity with different strains) have been used to identify critical amino acids of conformational epitopes [15, 16]. However, other critical amino acids of the conformational epitope with neutralizing activity against PCV2 capsid protein have not been identified. In this study, one mAb against the capsid protein of PCV2 was produced and characterized. Meanwhile, one key amino acid constituent of the conformational epitope was identified by using chimeras and mutants of PCV2a/CL and PCV2b/YJ strains.