Rita de Cássia Compagnoli Carmona

Human Rotavirus Serotype G9, São Paulo, Brazil, 1996-2003

Diverse rotavirus strains are present, and frequency of G9 is high.

Characterization of rotavirus strains from hospitalized and outpatient children with acute diarrhoea in São Paulo, Brazil

From August 1994 to July 1995, 234 faecal samples from children with or without acute diarrhoea were collected and tested. The group of children with acute diarrhoea (A) was subdivided into two subgroups: subgroup A1 was made up of children with severe diarrhoea, dehydrated and who needed hospitalization and subgroup A2 was composed of children who only needed outpatient care. Group B was composed of children without acute diarrhoea (controls). Rotavirus was detected in 36.7% (18/49), 22.0% (15/68) and 1.7% (2/117) patients in groups A1, A2 and B, respectively. Of the 35 positive samples in which rotaviruses were detected the VP7 genotypes G1, G2, G3, G5 and the mixture (G2 + G5) were found in 40.0, 11.4, 11.4, 22.9 and 2.9% of the samples, respectively. Also, the VP4 genotypes P[8], P[4] and P[6] were detected in 57.1, 31.4 and 5.7%, respectively. Rotavirus VP6 subgroups I and II were detected at a frequency of 22.4 and 54.3%, respectively. Rotavirus RNA segments had short and long electrophoresis profiles in 20.0 and 51.4% of the cases, respectively. The severity of the disease was not related to a specific G and P types, subgroup or electropherotype. J. Med. Virol. 74:166–172, 2004.

Rare G3P(3) rotavirus strain detected in Brazil: Possible human-canine interspecies transmission

BACKGROUND An unusual strain of human rotavirus G3P[3] (R2638 strain) was detected from a 1-year-old child patient during the epidemiological survey of rotavirus in the state of São Paulo, Brazil in 2011. OBJECTIVE The aim of this study was to carry out sequence analyses of the two outer capsid proteins (VP4 and VP7) of the R2638 strain detected in order to obtain further information of the genetic relationships between human and animal rotaviruses. STUDY DESIGN Rotavirus G3P[3] was detected using a commercial immunoenzymatic assay, SDS-PAGE, and genotyped by RT-PCR. The analysis of the genetic relationship between human and animal rotaviruses was carried out by sequencing the VP7 and VP4 genes. RESULTS The VP7 gene of the R2638 strain displayed the highest nucleotide identity to the canine strains A79-10 (96.6%) and CU-1 (96.2%) isolated in USA. The VP4 sequence showed the highest nucleotide identity to P[3] canine rotavirus strain RV52/96 isolated in Italy at 94.1%. Furthermore, the VP4 genes of P[3] strains could be discriminated into two phylogentically distinct clusters. CONCLUSION The present study reinforces the hypothesis that animals rotaviruses might be able to cross the species barriers, and the lack of systematic surveillance of rotavirus infection in small animals hinders the ability to establish firm epidemiologic connections. Moreover, in 2006 rotavirus vaccine was included in the Brazilian Immunization Program, and selective vaccine pressure could increase the circulation of uncommon strains. This is the first report of G3P[3] in over 20-year period of monitoring in Brazil.

Norovirus 3rd Generation kit: an improvement for rapid diagnosis of sporadic gastroenteritis cases and valuable for outbreak detection.

The aim of this study was to evaluate the sensitivity and specificity of the Ridascreen(®) Norovirus 3rd Generation kit compared to the RT-PCR. A retrospective, descriptive study was conducted with 245 specimens from sporadic cases and outbreak surveillance samples of gastroenteritis in Brazil from 2006 to 2009. Overall, the kit showed a sensitivity of 61.8% and a specificity of 92.5%. The sensitivity for outbreaks diagnosis was 87.9% and specificity 83.8%. The Ridascreen(®) 3rd Generation could detect specimen containing genogroup (G) II with high sensitivity. However, GI and mixed infections (GI/GII) were unlikely to be detected by the kit. ELISA for Norovirus (NoV) detection provides a rapid, technically simple assay system that can be used to increase the surveillance of gastroenteritis outbreaks, especially in Public Health Laboratories with high sample throughput. This assay is useful for the detection of NoV outbreaks and is an improvement as compared to previous ELISAs; however, due to its sensitivity, RT-PCR in still required for routine NoV detection in sporadic cases investigation.

Characterization of rotavirus strains from day care centers: pre- and post-rotavirus vaccine era

OBJECTIVES In 2006 the rotavirus vaccine was included in the Brazilian Immunization Program. The aim of this study was to report the results of a 5-year surveillance study of rotavirus strains in children < 5 years with acute gastroenteritis from day care centers in the state of São Paulo, Brazil. METHODS This retrospective study was conducted with 30 day care centers from 2004 to 2008 with convenient surveillance fecal specimens, investigated by ELISA, SDS-PAGE, RT-PCR and gene sequencing to genotype characterization. RESULTS Rotavirus infection was detected in 28.3% of samples (38/134). The most frequent genotypes detected were G9P[8] and G1P[8] in 2004; G1P[8] in 2005; GNTP[NT] in 2006; G2P[4] in 2007; and there were no cases in 2008. Mixed infections were not observed. Detection rate declined from 65.7% (23/35) in 2004 to 50% (9/18) in 2007. CONCLUSIONS Genotype distribution varied according to collection year, accompanied by a reduction in detection rate. Use of rotavirus vaccine requires implementation of post-marketing surveillance to monitor rotavirus strain diversity and its efficacy against possible new emerging genotypes.

Enterovirus 71 infection and acute neurological disease among children in Brazil (1988–1990)

Surveillance for Enterovirus 71 (EV-71) infection in children up to 15 years of age was carried out in Brazil, from 1988 to 1990. Patients with acute neurological diseases (AND) such as flaccid paralysis, Bells palsy, acute cerebellar ataxia and Guillain-Barré syndrome were included in the study. EV-71 infection was detected in 24 of 426 children (5.6%) with AND. EV-71 infection was confirmed only by virus isolation in 13 children, by virus isolation and seroconversion in 4, and by seroconversion alone in 7. EV-71 was also isolated from 15 of the 427 household contacts (3.5%) of 165 AND patients. There was some evidence of high infectivity of EV-71: household clusters were detected in the case of 7 of 24 children (29.1%) infected with EV-71 and manifesting AND; EV-71 was isolated from 11/40 household contacts (27.5%) of the infected patients but from only 4/387 household contacts (1.0%) of children in whom it was not possible to demonstrate EV-71 infection. Seven of the 24 children infected with EV-71 exhibited residual motor deficiency when examined 6 months after the disease onset. The relevance of these results for the Plan for Global Eradication of Wild Poliovirus is discussed, as well as the need to increase knowledge about the behaviour of this virus and its possible association with AND.

Rotavirus in adults, Brazil, 2004–2011: G2P[4] dominance and potential impact on vaccination

OBJECTIVES The aim of this study was to monitor rotavirus (RV) infections in adults >18 years with acute gastroenteritis during 2004-2011 national Brazilian RV surveillance. In addition, to characterize the RV group A (RVA) strains in order to gain insight into the supposed vaccine selective pressure imposed to Brazilian children population. METHODS A total of 2102 convenient fecal specimens were investigated by ELISA, PAGE, and RT-PCR. RESULTS RV was detected in 203 (9.6%) of 2102 specimens, and showed a marked peak of detection in September. RVA infection was detected in 9.4% (197/2102) and RV group C (RVC) in 0.3% (6/2102). The most frequent genotypes detected in 2004 and 2005 were G9P[8] (38.5%; 5/13) and G1P[8] (54.5%; 6/11), respectively. The dominant genotype identified from 2006 to 2011 was G2P[4] (64.4%; 116/180). Detection rate varied during the 8-year period of the study from 0.7% to 12.9%. CONCLUSION The high detection rate of G2P[4] in adults provides further evidence that its dominance reflects the seasonality of RVA strains instead of the supposed selective advantage created by vaccination program. It also can be suggested that adult infections may serve as a reservoir to maintain RVA strains in childhood gastroenteritis. Considering the detection rate, the evident reduction of RVA frequency observed in children after vaccine introduction was not present in adults.

Caracterização molecular de cepas de rotavírus e norovírus: um estudo de 6 anos (2004-2009)

OBJECTIVE To monitor rotavirus (RV) and norovirus (NoV) infections in hospitalized children ≤ 5 years with acute gastroenteritis in the state of São Paulo, Brazil, during a 6-year period (2004- 2009). METHODS This retrospective study was conducted with 61 medical centers with convenient surveillance fecal specimens, investigated by enzyme-linked immunosorbent assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis, reverse polymerase chain reaction and sequencing to genotype characterization. RESULTS RV and NoV infections were detected in 29.6% (144/487) and 29.2% (26/89) of the samples, respectively. The most frequent RV genotypes detected were G9P[8] in 2004; G1P[8] in 2005; G9P[8] in 2006; and G2P[4] during 2007, 2008, and 2009. Detection rate declined from 36.3% (33/91) in 2004 to 4.2% (4/95) in 2009. NoV genogroup GII was found in 61.6% (16/26) of the samples, and GI in 11.5% (3/26). Mixed NoV-RV infections were observed in 2.2% (2/89) of the samples, involving GI+G9P[8] and GI+G2P[4] strains. CONCLUSIONS Genotype distribution varied according to collection year, accompanied by a reduction in detection rate. Use of RV vaccine requires implementation of post-marketing surveillance to monitor RV strain diversity and its efficacy against possible new emerging genotypes. NoVs have been increasingly identified as relevant etiological agents among hospitalized children and play an important role in the viral etiology of pediatric acute gastroenteritis in the state of São Paulo.

Identification and Molecular Characterization of Norovirus in São Paulo State, Brazil

Norovirus (NoV), previously called Norwalk-like virus, represents an important group of human pathogens associated with outbreaks of nonbacterial gastroenteritis. Epidemiological surveys of outbreaks have shown that the most important routes of transmission are person-to-person contacts and contaminated food and water, with the virus affecting adults and children. NoV is classified into genogroups, being genogroups GI, GII and GIV found in humans. In view of the high genetic diversity of NoV and the lack of information about this virus in Brazil, the aim of the present study was the molecular characterization of NoV isolated from diarrheic stool samples of patients from São Paulo State. In this study, 204 stool specimens collected during diarrhea outbreaks were analyzed by RT-PCR, and 12 were sequenced for genogroup confirmation. One-step PCR was performed in order to amplify the B region of ORF 1 using the MON 431, 432, 433 and 434 primer pool. From total, 32 (15.7%) stool specimens were positive for NoV genogroup GII. Comparison of the sequences of the PCR products to GenBank sequences showed 88.8% to 98.8% identity, suggesting the presence of genogroup GII in gastroenteritis outbreaks in São Paulo State.

An exanthematic disease epidemic associated with coxsackievirus B3 infection in a day care center

An epidemic of exanthematic illness in a day care center is described. Ten children aged 7 to 13 months were affected by the illness. The exanthem was characterized by nonconfluent macular or maculopapular lesions that appeared on the face, body and limbs. Fifty percent of the infected children had fever of up to 39 degrees C at the beginning of the disease. Coxsackievirus B3 (CB3) was isolated from the stool of one ill child. Paired serum samples were obtained from eight ill children and six of them presented seroconversion to CB3. Antibodies to CB3 were detected at titers higher than 16 in a single serum sample collected from the other two patients. Neutralizing antibodies to CB3 were detected in 71.0% of the contact children.