ABSTRACT
The role of group C rotaviruses as a cause of diarrhea was examined among children <17 years of age admitted to a Hospital in a suburban area of Buenos Aires, Argentina between 1997 and 2003. A total of 1,579 fecal samples were screened for group A (RVA) and C (RVC) rotaviruses by two in-house ELISA methods at Quilmes University (UNQ-ELISA). Samples positive, doubtful and negative by RVC specific UNQ-ELISA (n = 246) were examined further for RVC by another in-house ELISA (CDC-ELISA), electron microscopy, RT-PCR, nested PCR, and Southern hybridization. Sensitivity, specificity, and predictive values for each test were determined. While the sensitivity was comparable for the nested PCR and CDC-ELISA methods (82.5%), the molecular methods were slightly more specific. Poorly preserved particles were often seen in fecal samples, suggesting that degradation of RNA could be a factor influencing the performance of molecular methods. The incidence of RVC was estimated to be 3% without apparent differences among seasons. RVC infected patients had a significantly (P < 0.001) higher median age (6 years vs. 1 year) than those with RVA infection. Sequence of the RVC VP7 gene from six Argentinean strains and sequences reported previously in different countries showed high nucleotide (94.4-99.9%) sequence identities, indicating a high degree of conservation for human RVC VP7 genes among strains collected on five continents over a period of 17 years. These findings indicate that RVC is a significant cause of diarrhea and it is necessary to develop simple and sensitive serological methods for its detection.
Subject(s)
Diarrhea/virology , Rotavirus Infections/diagnosis , Rotavirus/classification , Rotavirus/isolation & purification , Adolescent , Antigens, Viral/analysis , Argentina , Blotting, Southern/methods , Child , Child, Preschool , Conserved Sequence , Enzyme-Linked Immunosorbent Assay/methods , Feces/virology , Hospitals , Humans , Infant , Infant, Newborn , Molecular Sequence Data , Polymerase Chain Reaction/methods , Predictive Value of Tests , RNA, Viral/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Rotavirus/genetics , Rotavirus/immunology , Rotavirus Infections/virology , Sensitivity and Specificity , Sequence Analysis, DNA , Sequence HomologyABSTRACT
The incidence of human group C rotavirus infections among children and adults in Buenos Aires was evaluated by enzyme linked immunosorbent assays (ELISA) based on recombinant group C VP6 protein (Cowden strain). A total of 976 stool samples taken from patients (ages 6 months to 15 years) with acute diarrhea were tested for the presence of group C rotavirus. Among these, only 10 (1.02%) were group C rotavirus positive by enzyme-linked immunosorbent assay (ELISA) confirmed by absorption with group C VP6 antibodies and by RT-PCR for both VP6 and VP7 genes. The average age (5.86 years) was significantly superior to that in group A-infected patients (1.63 years). Previous exposure to this virus was assessed by detecting specific IgG in sera taken from healthy individuals grouped by age. Of 844 sera tested, 425 (50.3%) were group C IgG positive by ELISA, confirmed by Western blot analysis. The rates of IgG positivity for group A and C rotaviruses during the first years of life indicated that infections with group C are frequent in older children (3-5 years), whereas group A infections are prevalent in infants and young children (6-18 months). This study shows that group C rotavirus infections in Argentine children occur later in life than group A and are relatively common in spite of the low detection rate of this virus.
