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Development and evaluation of a nested-PCR assay for Senecavirus A diagnosis

Author:
Feronato, Cesar, Leme, RaquelA., Diniz, JaquelineA., Agnol, AlaisMaria Dall, Alfieri, AliceF., Alfieri, AmauriA.
Source:
Tropical animal health and production 2018 v.50 no.2 pp. 337-344
ISSN:
0049-4747
Subject:
DNA primers, RNA, Senecavirus, adults, farms, genome, herds, livestock and meat industry, mortality, neonates, piglets, reverse transcriptase polymerase chain reaction, sequence analysis
Abstract:
Senecavirus A (SVA) has been associated with vesicular disease in weaned and adult pigs and with high mortality of newborn piglets. This study aimed to establish a nested-PCR assay for the routine diagnosis of SVA infection. Tissue samples (n = 177) were collected from 37 piglets of 18 pig farms located in four different Brazilian states. For the nested-PCR, a primer set was defined to amplify an internal VP1 fragment of 316 bp of SVA genome. Of the 37 piglets, 15 (40.5%) and 23 (62.2%) were positive for the SVA in the RT-PCR and nested-PCR assays, respectively. The SVA RNA was detected in 61/177 (34.5%) samples with the RT-PCR, while the nested-PCR assay showed 84/177 (47.5%) samples with the virus (p < 0.05). According to the herds, 11 (61.1%) and 16 (88.9%) of the 18 pig herds were positive for the SVA in the RT-PCR and nested-PCR assays, respectively. Nucleotide sequencing analysis revealed similarities of 98.7–100% among SVA Brazilian strains and of 86.6–98% with SVA strains from other countries. The nested-PCR assay in this study was suitable to recover the SVA RNA in biological specimens, piglets, and/or herds that were considered as negative in the RT-PCR assay, and is proposed for the routine investigation of the SVA infection in piglets, especially when other techniques are not available or when a great number of samples has to be examined.
Agid:
5889308