The 545 sequences obtained from the rotavirus query in BLASTn were analyzed in the ConSort ß program to construct fw-and rev-primers. The fw-primer (nt 1-23), 5 0 -G GCTTTW+AAA+CGAA+GTC+TTCR-3 0 (+A, +C, +G, +T are LNA residues) and the biotinylated rev-primer (nt 502-426), 5 0 -TATGGAAATATATTAGG TTTATGAAAAACAAATCCIGTACGTTGTCTTCT ITTITGIARRTTCCAITTITCIATRTA-3 0 , resulted in a PCR product of 502 nt. After nucleic acid extraction in the easyMag Õ , the extracts were heated at 97 C for 5 min, followed by snap cooling on ice for 2 min to obtain ssRNA from the rotaviral dsRNA. The PCR reaction contained 5 ml of nucleic acid extract, 25 ml 2Â RT-PCR iScript buffer, 200-nM Fw-primer, 600-nM biotinylated rev-primer, nuclease-free water and 1 ml iScript RT enzyme (total volume 50 ml). The samples were run at 50 C for 30 min, 94 C for 10 min, 50 cycles at 94 C for 30 s, 55 C for 30 s and 72 C for 30 s, ending with 72 C for 7 min. An amount of 5 ml of the PCR product was used in the hybridization experiment with the microsphere-coupled Rotavirus probes (sequences shown in Supplementary Table S4A and B. The PCR products were sequenced with a 3130 Genetic Analyzer (Applied Biosystem) by utilizing the same fw-and rev-primers described above.