Molecular analysis of norovirus in specimens from children enrolled in a 1982-1986 study in Belém, Brazil: A community-based longitudinal study.
Fecal specimens were collected during a longitudinal, community-based study in the city of Belém, North Brazil, that was conducted over 3 years (October 1982 to March 1986), in which 20 children were included from birth to 3 years of age. A total of 229 fecal samples were screened by real time RT-PCR targeting the junction region (ORF 1/2) of the norovirus (NoV) genome. NoV-positive samples were subjected to PCR and sequencing of the viral polymerase (ORF1) and viral protein 1 (VP1) genes (ORF2). The junction region was also sequenced to assess for recombination when ORF1 and ORF2 genotyping results were dissimilar. Samples classified as GII.P4/GII.4 were further characterized by sequencing the P2 subdomain of the viral capsid to determine possible alterations. An overall positivity of 16.1% (37/229) was observed, including GI (16.2%-6/37) and GII (83.8%-31/37) genogroups. Cases of NoV reinfection in at least 2-month intervals were observed, and 12 children developed at least one case of asymptomatic NoV infection. In total, 48.6% (18/37) NoV-positive samples were subjected to nucleotide sequencing analysis targeting the following polymerase genes: GI.P3 (n = 1), GII.Pa (n = 1), GII.Pc (n = 1), GII.P4 (n = 5), GII.P6 (n = 5), GII.P7 (n = 3), GII.P12 (n = 1), and GII.P22 (n = 1). For the VP1 gene, characterization was performed in 14 (77.8%) samples: GI.3 (n = 1), GII.2 (n = 1), GII.4 (n = 4), GII.6 (n = 4), GII.7 (n = 1), GII.12 (n = 1), GII.14 (n = 1), and GII.23 (n = 1). Recombination events were confirmed in three cases (GII.P12/GII.2, GII.P7/GII.14, and GII.Pa/GII.12), and four samples genotyped as GII.P4/GII.4 were analyzed to identify variants. None had contemporary counterparts. Three children developed consecutive NoV infections by different genotypes. The present report documents the importance of NoV as a cause of childhood infection during a longitudinal study conducted more than 30 years ago.
Postgraduate Program in Virology, Evandro Chagas Institute. Ananindeua, Pará, Brazil.
Jones Anderson Monteiro
JAM
Siqueira
Postgraduate Program in Virology, Evandro Chagas Institute. Ananindeua, Pará, Brazil.
Edivaldo Costa Sousa
ECS
Júnior
Virology Section, Evandro Chagas Institute, Health Surveillance Secretariat, Brazilian Ministry of Health. Ananindeua, Pará, Brazil.
Alexandre da Costa
ADC
Linhares
Virology Section, Evandro Chagas Institute, Health Surveillance Secretariat, Brazilian Ministry of Health. Ananindeua, Pará, Brazil.
Yvone Benchimol
YB
Gabbay
D001938:Brazil / Q000453:epidemiology; D017250:Caliciviridae Infections / Q000453:epidemiology / Q000821:virology*; D036022:Capsid Proteins / Q000235:genetics; D002675:Child, Preschool; D005243:Feces / Q000821:virology*; D005260:Female; D005759:Gastroenteritis / Q000453:epidemiology / Q000821:virology*; D014644:Genetic Variation; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008137:Longitudinal Studies; D008297:Male; D029322:Norovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016366:Open Reading Frames; D010802:Phylogeny; D011634:Public Health; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D011995:Recombination, Genetic
D036022:Capsid Proteins; D012367:RNA, Viral
children; diarrhea; gastroenteritis; norovirus