Quantification and Genotyping of Human Sapoviruses in the Llobregat River Catchment, Spain

Abstract

Human sapoviruses (SaVs) were quantified and characterized in an 18-month survey conducted along the Llobregat river catchment area in Spain. Sample types included freshwater, untreated and treated wastewater, and drinking water. All genogroups were recovered, and a seasonal distribution was observed. This is the first report of SaV quantification and genotyping in the environment outside Japan.

Diverse sapovirus genotypes identified in children hospitalised with gastroenteritis in selected regions of South Africa

“…Strains from SA shared even higher nucleotide identity (99-100%) with SaVs from other countries, particularly recent strains reported in Asia (2012) [18] and Japan (2003) [5]. In GII.5, the most closely-related SaV (93-95% nucleotide identity) was identified from wastewater in Japan (2005) [22] and in GII.6 GenBank matches with < 90% nucleotide identity over the genotyped region included SaVs detected in stool from Burkina Faso [14], Thailand [37] and Japan and river water in Spain [23], from 2004 to 2009. In GII.7, closely-related SaVs from other countries included recent strains from the US (2010) and China (2011) [38].…”

“…The region was amplified by nested polymerase chain reaction (PCR) using published primers [22,23] as previously described [24], with minor adjustments. Briefly, in the first round of PCR 5 µl cDNA was added to a 50 µl reaction with 0.4 µM of each primer (SV-F13, SV-F14, SV-DS3 and SV-DS4) and 1.25 U AmpliTaq Gold DNA polymerase (Applied Biosystems, Foster City, CA), with cycling parameters as previously described [24].…”

Application of a Competitive Internal Amplification Control for the Detection of Sapoviruses in Wastewater

“…Likewise, in the northern hemisphere SaV concentrations were at their lowest or not detected in the summer months of July and August (Haramoto et al 2008;Sano et al 2011). …”

“…and environmental samples including river water (Kitajima et al 2010;Sano et al 2011), wastewater (Haramoto et al 2008;Kitajima et al 2011) and shellfish (Hansman et al 2007b;Iizuka et al 2010;Ueki et al 2010); predominantly in J a p a n a n d S p a i n . H o w e v e r , w h e n c o m p a r e d w i t h N o V s , t h e r e i s l i m i t e d information available on SaVs circulating in the environment worldwide.…”

“…Sapoviruses are most frequently detected in environmental samples using realtime reverse transcription-quantitative polymerase chain reaction (RT-qPCR) (Haramoto et al 2008;Kitajima et al 2010;Sano et al 2011). This molecular method allows for sensitive and specific detection and quantification of SaVs (Chan et al 2006) but its application can be hampered by the presence of inhibitory compounds in the sample.…”