Genetic Diversity of Norovirus among Children with Gastroenteritis in São Paulo State, Brazil

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Abstract
Norovirus (NoV) is one of the most common causes of acute gastroenteritis in children and adults. To study the prevalence and genetic variability of NoV in children with acute gastroenteritis in São Paulo State, Brazil, we examined 234 stool samples from children with or without gastroenteritis during a 5-year period (1995 to 1999). NoV RNA was detected by reverse transcription-PCR and confirmed by DNA sequence analysis. We used two different oligonucleotide primer sets targeting the 3′ end of the RNA polymerase gene (region B), as well a partial capsid region at the 3′ end of the VP1 gene (region D). A total of 78 (33.3%) of the samples tested positive for NoV, and in region B, of the 66 strains sequenced, 4 (6.1%) belonged to GI, 52 (78.7%) belonged to GII, and five samples (7.6%) contained a mixture of the GI and GII genotypes. Phylogenetic analysis showed that the majority (40 of 66 [60.6%]) of the strains belonged to genotype GII.4. The nucleotide sequence identity of three strains was lower than 77.9% compared to a region B reference sequence database but showed 85.3 to 88.8% identity with GII.2 Melksham strain in region D, indicating the circulation of a possible recombinant NoV strain. One sample (GII.3) was sequenced only in region D. In conclusion, we have a total of 67 sequenced strains. This is the first report that describes the predominance of GII.4 NoV strains in children visiting the ambulatory of different hospitals in São Paulo State, Brazil, and we show that mixtures of different strains can be found in individual samples, including some possible new recombinant strains.

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