Abstract
BackgroundSapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae. The virus is detected in different species including the human and the porcine species as an enteric pathogen causing asymptomatic to symptomatic enteritis. In this study, we report the development of a rapid real time qRT-PCR based on SYBR Green chemistry for the diagnosis of porcine sapovirus infection in swine.ResultsThe method allows the detection of porcine sapoviruses and the quantification of the genomic copies present in stool samples. During its development, the diagnostic tool showed good correlation compared with the gold standard conventional RT-PCR and was ten-fold more sensitive. When the method was applied to field samples, porcine noroviruses from genogroup 2 genotype 11b were also detected. The method was also applied to swine samples from the Netherlands that were positive for PoSaV infection. Phylogenetic results obtained from the samples showed that PoSaV sequences were genetically related to the currently described genogroup III, to the proposed genogroup VII and also to the MI-QW19 sequence (close to the human SaV sequences).ConclusionsA rapid, sensitive, and reliable diagnosis method was developed for porcine sapovirus diagnosis. It correlated with the gold standard conventional RT-PCR. Specificity was good apart for genogroup 2 genotype 11b porcine noroviruses. As a first line screening diagnosis method, it allows a quicker and easier decision on doubtful samples.
Highlights
Sapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae
The present study shows that when the gold standard conventional RT-PCR was applied to the field samples, discrimination between amplicons found near the expected molecular weight and the right amplicons was sometimes difficult in comparison with the easier analysis allowed by melting curve analysis as part of the SYBR green qRT-PCR
Porcine sapoviruses (PoSaV) diagnosis, currently based on conventional RT-PCR, requires a powerful, rapid, sensitive, specific and reliable tool, in order to investigate the epidemiological situation in Europe
Summary
Sapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae. Sapoviruses (SaV) belong to the genus Sapovirus within the family Caliciviridae These small (~30 nm), icosahedric, non-enveloped viruses have a single-stranded, positive sense, RNA genome. In Europe, PoSaV sequences were recently detected in about 50% of tested samples from six countries (Denmark, Spain, Slovenia, Italy, Finland and Hungary) with a molecular prevalence of 7.6% in all the samples [7]. Alongside this situation of rising levels of SaVs in the veterinary world lies the human context where, in Europe, SaV infections are being increasingly reported [22]. These data highlight the need for rapid, sensitive, specific and cost effective diagnostic tools to detect PoSaV infection
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