Development and application of a SYBR green RT-PCR for first line screening and quantification of porcine sapovirus infection.
BACKGROUND
Sapoviruses 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.


RESULTS
The 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).


CONCLUSIONS
A 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.
Veterinary Virology and Animal Viral Diseases, Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. amauroy@ulg.ac.be
Axel
A
Mauroy
Wim H M
WH
Van der Poel
Renate Hakze-Van
RH
der Honing
Christine
C
Thys
Etienne
E
Thiry
D000818:Animals; D001483:Base Sequence; D017250:Caliciviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D005456:Fluorescent Dyes; D005759:Gastroenteritis / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016679:Genome, Viral; D005838:Genotype; D006801:Humans; D010802:Phylogeny; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D029341:Sapovirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology*
D005456:Fluorescent Dyes