Identification and Characterization of Porcine Kobuvirus Variant Isolated from Suckling Piglet in Gansu Province, China
Kobuviruses comprise three species, the Aichivirus A, Aichivirus B, and Aichivirus C (porcine kobuvirus). Porcine kobuvirus is endemic to pig farms and is not restricted geographically but, rather, is distributed worldwide. The complete genomic sequences of four porcine kobuvirus strains isolated during a diarrhea outbreak in piglets in the Gansu province of China were determined. Two of these strains exhibited variations relative to the traditional strains. The potential 3C/3D cleavage sites of the variant strains were Q/C, which differed from the Q/S in the traditional porcine kobuvirus genome. A 90-nucleotide deletion in the 2B protein and a single nucleotide insertion in the 3′UTR were found in the variant strains. The VP1 regions of all four porcine kobuviruses in our study were highly variable (81%–86%). Ten common amino acid mutations were found specifically at certain positions within the VP1 region. Significant recombination sites were identified using SimPlot scans of whole genome sequences. Porcine kobuviruses were also detected in pig serum, indicating that the virus can escape the gastrointestinal tract and travel to the circulatory system. These findings suggest that mutations and recombination events may have contributed to the high level of genetic diversity of porcine kobuviruses and serve as a driving force in its evolution.
[…] PCR products were cloned into the pMD-18 T vector (TaKaRa, Otsu, Japan). Competent E. coli DH5a cells (TaKaRa) were transformed with the recombinant plasmid. Three clones were sequenced using the Applied Biosystems (ABI) 3730xl DNA analyzer. Sequences were analyzed using the ClustalX version 2.0 , DNASTAR, and MegAlign version 5.0 (DNAStar Inc., Madison, WI, USA) software packages. A dendrogram was constructed using the neighbor-joining clustering method in MEGA 5.0 . Possible recombination events in the coding region were analyzed using Simplot software . All kobuvirus strains used in the study are listed in . […]
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