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These samples had all previously been tested for rotaviruses [Kiulia et al., 2009], astroviruses [Kiulia et al., 2007], and adenoviruses (AdVs) [Magwalivha et al., 2010]. Stool suspensions (10%) were prepared in ultrapure water and stored at ?20��C until nucleic acid extraction. Total nucleic acids were extracted from 200??l stool suspension using the MagNA Pure LC Total Nucleic Acid Isolation kit (Roche Diagnostics, Mann-heim, Germany) on the automated MagNA Pure system (Roche Diagnostics). The nucleic acids were eluted in 50??l and stored at ?70��C until use. NoV GI and GII were detected with published one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assays targeting the ORF1/ORF2 junction [Mans et al., 2010]. Specimens were screened for NoV GIV [Trujillo et al., 2006; Murray et al., 2013a] and SaV [Chan et al., 2006; Murray et al., 2013b] as previously described. NoV GI and BLZ945 cost GII strains were genotyped based on nucleotide sequence determination and phylogenetic analysis of the 5��-end of the capsid gene (Region C) using a semi-nested RT-PCR as described previously [Mans et al., 2013]. Briefly, a first round of amplification was performed with primer pairs QNIF4/G1SKR for NoV GI and QNIF2/G2SKR for NoV GII. If no PCR products were obtained after this step, a second amplification was performed using primers G1SKF/G1SKR and G2SKF/G2SKR. BMS-754807 manufacturer SaVs were genotyped based on partial capsid gene nucleotide sequences (approximately 300?bp) as described previously [Kitajima et al., 2010; Sano et al., 2011; Murray et al., 2013a]. The PCR products were purified with the DNA Clean and Concentrater kit (Zymo Research, Irvine, CA) and directly sequenced with the ABI PRISM BigDye? Terminator v. 3.1 Cycle Sequencing kit on an ABI 3130 automated analyzer (Applied Biosystems, Foster City, CA). Nucleotide sequences were edited and analyzed using Sequencher? 4.9 (Gene Codes Corporation, Ann Arbor, MI) and BioEdit Sequence Alignment Editor (V.7.0.9.0) [Hall, 1999]. Phylogenetic analysis of NoV GI, GII, and SaV was performed in MEGA5 using the neighbor-joining method, validated by 1,000 bootstrap replicates as described previously [Murray et al., 2013a]. Genotypes were assigned based on clustering with reference strains in the phylogenetic tree with >70% bootstrap INSRR support. The NoV Genotyping Tool [Kroneman et al., 2011] was used to confirm the NoV genotype assignment. Nucleotide sequences determined in this study were submitted to GenBank under accession numbers: KF279373-KF279391 (NoV) and KF267740-KF267745 (SaV). Statistical significance was determined by calculating a 2?��?2 contingency table using the Fischer's Exact test with Graphpad Quickcalcs (www.graphpad.com/quickcalcs/contingency2/). P-Values