Pegivirus Detection in Cerebrospinal Fluid from Patients with Central Nervous System Infections of Unknown Etiology in Brazil by Viral Metagenomics.

Metagenomic next-generation sequencing (mNGS) methodology serves as an excellent supplement in cases where diagnosis is challenging to establish through conventional laboratory tests, and its usage is increasingly prevalent. Examining the causes of infectious diseases in the central nervous system (CNS) is vital for understanding their spread, managing outbreaks, and effective patient care. In a study conducted in the state of São Paulo, Brazil, cerebrospinal fluid (CSF) samples from 500 patients with CNS diseases of indeterminate etiology, collected between 2017 and 2021, were analyzed. Employing a mNGS approach, we obtained the complete coding sequence of Pegivirus hominis (HPgV) genotype 2 in a sample from a patient with encephalitis (named IAL-425/BRA/SP/2019); no other pathogen was detected. Subsequently, to determine the extent of this virus's presence, both polymerase chain reaction (PCR) and/or real-time PCR assays were utilized on the entire collection. The presence of the virus was identified in 4.0% of the samples analyzed. This research constitutes the first report of HPgV detection in CSF samples in South America. Analysis of the IAL-425 genome (9107 nt) revealed a 90% nucleotide identity with HPgV strains from various countries. Evolutionary analyses suggest that HPgV is both endemic and extensively distributed. The direct involvement of HPgV in CNS infections in these patients remains uncertain.

First Detection of DS-1-like G1P[8] Double-gene Reassortant Rotavirus Strains on The American Continent, Brazil, 2013.

Emergence of DS-1-like-G1P[8] rotavirus in Asia have been recently reported. We report for the first time the detection and the whole genome phylogenetic analysis of DS-1-like-G1P[8] strains in America. From 2013 to 2017, a total of 4226 fecal samples were screened for rotavirus by ELISA, PAGE, RT-PCR and sequencing. G1P[8] represented 3.7% (30/800) of all rotavirus-positive samples. DS-1-like-G1P[8] comprised 1.6% (13/800) detected exclusively in 2013, and Wa-like-G1P[8] comprised 2.1% (17/800) detected from 2013 to 2015. Whole genome sequencing confirmed the DS-1-like backbone I2-R2-C2-M2-A2-N2-T2-E2-H2. All genome segments of the Brazilian DS-1-like-G1P[8] strains clustered with those of Asian strains, and apart from African DS-1-like-G1P[8] strains. In addition, Brazilian DS-1-like-G1P[8] reassortants distantly clustered with DS-1-like backbone strains simultaneously circulating in the country, suggesting that the Brazilian DS-1-like-G1P[8] strains are likely imported from Asia. Two distinct NSP4 E2 genotype lineages were also identified, indicating the existence of a co-circulating pool of different DS-1-like G1P[8] strains. Surveillance systems must be developed to examine if RVA vaccines are still effective for the prevention against unusual DS-1-like-G1P[8] strains.

Spread of the emerging equine-like G3P[8] DS-1-like genetic backbone rotavirus strain in Brazil and identification of potential genetic variants.

In 2013, the equine-like G3P[8] DS-1-like rotavirus (RVA) strain emerged worldwide. In 2016, this strain was reported in northern Brazil. The aims of the study were to conduct a retrospective genetic investigation to identify the possible entry of these atypical strains in Brazil and to describe their distribution across a representative area of the country. From 2013 to 2017, a total of 4226 faecal samples were screened for RVA by ELISA, PAGE, RT-PCR and sequencing. G3P[8] represented 20.9 % (167/800) of all RVA-positive samples, further subdivided as equine-like G3P[8], DS-1-like (11.0 %; 88/800) and Wa-like G3P[8] (9.9 %; 79/800). Six equine-like G3P[8] DS-1-like samples were selected for whole-genome investigation, confirming the backbone I2-R2-C2-M2-A2-N2-T2-E2-H2. During 2013-2014, Wa-like G3P[8] was predominant and no equine-like G3P[8] DS-1-like was detected. Equine-like G3P[8] DS-1-like was first identified in Paraná in March/2015, suggesting that the strain entered Brazil through the Southern region. Equine-like G3P[8] rapidly spread across the area under surveillance and displayed a marked potential to replace Wa-like G3P[8] strains. Brazilian equine-like G3P[8] DS-1-like strains clustered with contemporary equine-like G3P[8] DS-1-like detected worldwide, but exhibited a distinct NSP2 genotype (N2) compared to the previously reported Amazon equine-like G3P[8] DS-1-like strain (N1). Two distinct NSP4 E2 genotype lineages were also identified. Taken together, these data suggest that different variants of equine-like G3P[8] DS-1-like strains might have been introduced into the country at distinct time points, and co-circulated in the period 2015-2017. The global emergence of equine-like G3P[8] DS-1-like strains, predominantly in countries using the Rotarix vaccine, raises the question of whether vaccines may be inducing selective pressures on zoonotic strains.

Detection and genetic characterization of classic human astroviruses in Brazil, 2010-2012.

The aims of this study were to monitor human astrovirus (HAstV) infections in patients presenting with acute gastroenteritis in Brazil and to determine the HAstV genotypes of these viruses. From May 2010 to July 2012, a total of 140 samples that were negative for both rotaviruses and noroviruses were randomly selected and tested for the presence of HAstV using an RT-PCR assay specific for the ORF2 region. Viral genotypes were identified and genetic diversity was investigated by sequencing. HAstV infection was detected in 2.9% of samples (4/140). The viruses in three samples were shown by phylogenetic analysis to belong to HAstV-4 lineage "c", clustering together with strains detected in Europe and the Middle East. The virus in one sample was genotyped as HAstV-1 lineage "a", clustering with strains from Uruguay, Brazil and Russia. Our findings provide further evidence for a global distribution of HAstV-1a and suggest a possible emergent importance of the HAstV-4c lineage in this country. The present study does not suggest that HAstVs currently have a major epidemiological impact, even after the introduction of a rotavirus vaccine in 2006.

Rotavirus genotypes as etiological agents of diarrhoea in general populations of two geographic regions of Brazil.

Rotavirus is the main global cause of severe childhood diarrhoea among children. In 2006, Rotarix® (G1P[8]) was introduced into Brazil's National Immunization Program. The vaccine coverage rate was 84.4% in 2009. Evidences of increasing G2P[4] after 2006 opened up the discussion about the vaccine effectiveness to non-G1 strains. The aim of this study was to identify the circulating rotavirus genotypes in two Brazilian regions during 2009. A total of 223 positive samples by immunochromatography and latex agglutination assay from the Northeast (Bahia/Pernambuco States) and Southeast (São Paulo/Rio de Janeiro States) regions were included in the study. The samples were submitted to genotyping by nested-PCR according to VP7(G) and VP4(P) and 175 samples (78.5%) were able to be characterized. Considering the characterization of VP7, the G-types detected were G1, G2, and G4 in the Northeast, and G2, G3, G5, and G9 in the Southeast. Considering the characterization of VP4, the P-types detected were P[4], P[8], and P[6]/P[9] in the Northeast and the Southeast. The most frequent mixed types found were G2P[4]/G2P[NT](81.4%), G2P[6](5.2%), G1P[6](5.2%) in the Northeast, and G2P[4]/G2P[NT](78.8%), G2P[6](8.2%), G9P[8](4.7%) in the Southeast. Among immunized individuals whose age ranged from 0-4 years, the G2P[4]/G2P[NT] genotype was identified in 91,0% of cases, and among non-immunized individuals of the same age, the G2P[4]/G2P[NT] genotype was identified in 85.7% of the cases. In accordance with the high level of vaccine coverage, the data suggest that the circulation of G2P[4] in these regions had a considerable increase after the introduction of Rotarix®.

Outbreak of G2P[4] rotavirus gastroenteritis in a retirement community, Brazil, 2015: An important public health risk?

The present study described a group A rotavirus (RVA) outbreak in an age-care facility in Brazil, using epidemiologic and molecular diagnostic methods. A descriptive clinical, epidemiological and environmental investigation was conducted. Stool samples were collected and screened for RVA, Norovirus (NoV), Enteric Adenovirus 40/41 (AdV 40/41) and Astrovirus (AstV) using ELISA, RT-PCR, qRT-PCR, electron microscopy and sequencing methods. Outbreak occurred during 26th-29th October, 2015; 28 individuals affected (22 residents; 6 staff). The attack rate was 25.9% and 8.5% among residents (median-age: 85.5 years) and staff (median-age: 28 years), respectively. Female staff was identified as the index case. RVA G2P[4] genotype was detected in 87.5% (7/8). Genetic analysis demonstrated that the outbreak involved one single strain, suggesting a common-source infection. RVA should be considered during outbreaks investigations in residential facilities, and raise the question if the current licensed RVA vaccines for children could also be helpful for the elderly.

Detection of the emerging rotavirus G12P[8] genotype at high frequency in brazil in 2014: Successive replacement of predominant strains after vaccine introduction.

The continuum characterization of rotavirus (RVA) genotypes is essential to understand how vaccine introduction could impact virus epidemiology. In the present study, an unexpected rapid changing pattern of RVA genotypes distribution in Brazilian population during three followed seasons is described. From January/2012 to December/2014, a total of 3441 fecal specimens were collected from collaborating centers across Southern, Southeastern and Midwest of Brazil. All specimens were screened for RVA using ELISA, and genotyped by RT-PCR. Differences in proportions were tested using Chi-Squares. A p-value of less than 0.05 was considered statistically significant. RVA was detected in 19.7% (677/3441). Among RVA positive cases (n=677), a total of 652 (96.3%) samples were successfully amplified by RT-PCR. G3P[8] remained prevalent in 2012 (37.6%, 69/185) and 2013 (40.1%, 74/186) (χ(2)=0.107, p=0.743), but declined markedly in 2014 (3.5%, 10/281) (χ(2)=71.770, p=0.000). G12P[8] was second highest strain in 2012 (22.7%, 42/185), decrease rapidly in 2013 (2.7%, 5/186) (χ(2)=26.224, p=0.000) and re-emerged as the predominant genotype in 2014 (86.6%, 243/281) (χ(2)=118.299, p=0.000). From July/2014, G12P[8] was the single genotype detected in all regions studied. The sudden emergence, spread and predominance of G12P[8] strain in Brazil, raised the hypothesis of a possible G12 outbreak being in progress. Nationally, the long term decline in gastroenteritis hospitalization observed in the country after RVA vaccine introduction was confirmed. Nevertheless, the sharp increase in diarrhea hospitalization prevalence from 2013 to 2014 observed in Southern and Southeastern regions is consistent with what appears to be an outbreak of G12P[8]. Continued surveillance is needed to verify the effectiveness of the RotarixTM vaccine in Brazil together with potential emergence of unusual genotypes.

Avaliação do desempenho da PCR em tempo real para o diagnóstico de meningite por Enterovirus / Evaluation of a real-time PCR assay performance for the diagnosis of enteroviral meningitis

O gênero Enterovirus (EV) é o agente etiológico mais frequente e responsável pela ocorrência de meningite viral no mundo. O objetivo deste trabalho foi de avaliar resultados da implantação do ensaio de PCR em tempo real (RT-qPCR) para a detecção de EV. Foram selecionadas 616 amostras de líquido cefalorraquidiano (LCR) de pacientes com meningite, recebidas para realizar diagnóstico laboratorial no período de 1998-2013. Os RNAs foram extraídos diretamente do LCR pelo método QIAamp®, e o ensaio TaqMan® foi aplicado. A avaliação foi feita comparando-se resultados de RT-qPCR com os obtidos pelo método de isolamento em cultura de células. Das 616 amostras analisadas, 94 (15,2 %) foram positivas para EV no ensaio de RT-qPCR; e na cultura celular EV foi isolado de 58 (9,4 %) amostras. Valores de sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo foram de 89,70 %, 92,40 %, 55,30 % e 98,90 %, respectivamente. O RT-qPCR foi ligeiramente superior à cultura viral para a detecção de EV no LCR. O RT-qPCR TaqMan® é um ensaio rápido e sensível, facilmente exeqüível e com potencial para melhorar o diagnóstico da meningite viral na rotina do laboratório de saúde pública no Estado de São Paulo.

Enterovirus (EV) genus is the most frequent etiological agent causing viral meningitis worldwide. This study aimed at evaluating the performance of real-time reverse transcription-PCR (RT-qPCR) assay for detecting EV. A total of 616 cerebrospinal fluid (CSF) samples from patients with meningitis were selected, among those received at EV diagnosis laboratory from 1998 to 2013. RNAs were directly extracted from CSF by using QIAamp® Viral RNA Mini Kit, and TaqMan® RT-qPCR assay was applied. Evaluation was made by comparing the RT-qPCR results with those found in the cell culture for viral isolation method. Of 616 analyzed samples, 94 (15.20%) were positive for EV RNA on the RT-qPCR assay; and in the cell culture EV was isolated from 58 (9.40 %) samples. The assay showed sensitivity, specificity, positive predictive value and negative predictive value of 89.70 %, 92.40 %, 55.30 % and 98.90 %, respectively. In the present study, the RT-qPCR assay was slightly superior when compared to the viral culture technique for detecting EV from CSF samples. The TaqMan® RT-qPCR assay shows to be a fast and sensitive assay, easy to perform, and it shows a potential to improve the viral meningitis diagnosis in the public health laboratory in São Paulo State.

Avaliação do desempenho da PCR em tempo real para o diagnóstico de meningite por Enterovirus / Evaluation of a real-time PCR assay performance for the diagnosis of enteroviral meningitis

O gênero Enterovirus (EV) é o agente etiológico mais frequente e responsável pela ocorrência de meningite viral no mundo. O objetivo deste trabalho foi de avaliar resultados da implantação do ensaio de PCR em tempo real (RT-qPCR) para a detecção de EV. Foram selecionadas 616amostras de líquido cefalorraquidiano (LCR) de pacientes com meningite, recebidas para realizar período de 1998-2013. Os RNAs foram extraídos diretamente do LCR pelo método QIAamp®, e o ensaio TaqMan® foi aplicado. A avaliação foi feita comparando-se resultados de RT-qPCR com os obtidos pelo método de isolamento em cultura de células.Das 616 amostras analisadas, 94 (15,2 %) foram positivas para EV no ensaio de RT-qPCR; e na cultura celular EV foi isolado de 58 (9,4 %) amostras. Valores de sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo foram de 89,70 %, 92,40 %, 55,30 % e 98,90 %,respectivamente. O RT-qPCR foi ligeiramente superior à cultura viral para a detecção de EV no LCR. O RT-qPCR TaqMan® é um ensaio rápido e sensível, facilmente exeqüível e com potencial para melhorar o diagnóstico da meningite viral na rotina do laboratório de saúde pública noEstado de São Paulo.

The genus Enterovirus (EV) is the most frequent etiological agent responsible for the occurrence of viral meningitis in the world. The objective of this work was to evaluate the results of the implantation of the real-time PCR assay (RT-qPCR) for the detection of EV. We selected 616 samples of cerebrospinal fluid (CSF) from patients with meningitis, received for the period 1998-2013. The RNAs were extracted directly from the CSF by the QIAamp® method, and the TaqMan® assay was applied. The results of the RT-qPCR test were compared with those obtained by the cell culture isolation method. Of the 616 samples analyzed, 94 (15.2%) were positive for EV in the RT-qPCR assay; And cell culture EV was isolated from 58 (9.4%) samples. Sensitivity, specificity, positive predictive value and negative predictive values ​​were 89.70%, 92.40%, 55.30% and 98.90%, respectively. The RT-qPCR was slightly superior to the viral culture for the detection of EV in the CSF. RT-qPCR TaqMan® is a rapid and sensitive, easily feasible and potential test to improve the diagnosis of viral meningitis in the routine of the public health laboratory in the State of São Paulo.

ROTAVIRUS GENOTYPES CIRCULATING IN BRAZIL, 2007-2012: IMPLICATIONS FOR THE VACCINE PROGRAM.

Regarding public health in Brazil, a new scenario emerged with the establishment of universal rotavirus (RV) vaccination programs. Herein, the data from the five years of surveillance (2007-2012) of G- and P-type RV strains isolated from individuals with acute gastroenteritis in Brazil are reported. A total of 6,196 fecal specimens were investigated by ELISA and RT-PCR. RVs were detected in 19.1% (1,181/6,196). The peak of RV incidence moved from June-August to September. RV was detected less frequently (19.5%) among children ≤ 5 years than in older children and adolescents (6-18 years) (40.6%). Genotype distribution showed a different profile for each year: G2P[4] strains were most prevalent during 2007-2010, G9P[8] in 2011, and G12P[8] in 2012. Mixed infections (G1+G2P[4], G2+G3P[4]+P[8], G2+G12P[8]), unusual combinations (G1P[4], G2P[6]), and rare strains (G3P[3]) were also identified throughout the study period. Widespread vaccination may alter the RV seasonal pattern. The finding of RV disease affecting older children and adolescents after vaccine implementation has been reported worldwide. G2P[4] emergence most likely follows a global trend seemingly unrelated to vaccination, and G12, apparently, is emerging in the Brazilian population. The rapidly changing RV genotype patterns detected during this study illustrate a dynamic population of co-circulating wildtype RVs in Brazil.

Rotavirus genotypes circulating in Brazil, 2007-2012: implications for the vaccine program / Genótipos de rotavírus circulando no Brasil, 2007-2012: implicações para o Programa de Vacinação

SUMMARYRegarding public health in Brazil, a new scenario emerged with the establishment of universal rotavirus (RV) vaccination programs. Herein, the data from the five years of surveillance (2007-2012) of G- and P-type RV strains isolated from individuals with acute gastroenteritis in Brazil are reported. A total of 6,196 fecal specimens were investigated by ELISA and RT-PCR. RVs were detected in 19.1% (1,181/6,196). The peak of RV incidence moved from June-August to September. RV was detected less frequently (19.5%) among children ≤ 5 years than in older children and adolescents (6-18 years) (40.6%). Genotype distribution showed a different profile for each year: G2P[4] strains were most prevalent during 2007-2010, G9P[8] in 2011, and G12P[8] in 2012. Mixed infections (G1+G2P[4], G2+G3P[4]+P[8], G2+G12P[8]), unusual combinations (G1P[4], G2P[6]), and rare strains (G3P[3]) were also identified throughout the study period. Widespread vaccination may alter the RV seasonal pattern. The finding of RV disease affecting older children and adolescents after vaccine implementation has been reported worldwide. G2P[4] emergence most likely follows a global trend seemingly unrelated to vaccination, and G12, apparently, is emerging in the Brazilian population. The rapidly changing RV genotype patterns detected during this study illustrate a dynamic population of co-circulating wildtype RVs in Brazil.

RESUMOEm relação à saúde pública no Brasil, um novo cenário emergiu com o estabelecimento dos programas universais de vacinação contra o rotavírus (RV). Os resultados de cinco anos (2007-2012) de vigilância dos genótipos G e P de cepas de RV detectadas em indivíduos com gastroenterite aguda no Brasil são descritos no presente estudo. Um total de 6196 amostras fecais foi investigado utilizando ELISA e RT-PCR. RVs foram detectados em 19,1% (1181/6196). O pico de incidência de RV se deslocou de junho-agosto para setembro. RV foi detectado com menor frequência entre crianças ≤ 5 anos (19,5%) quando comparado às crianças mais velhas e adolescentes (6-18 anos) (40,6%). A distribuição genotípica mostrou um perfil diferente a cada ano: a cepa G2P[4] foi prevalente durante 2007-2010, G9P[8] em 2011 e G12P[8] em 2012. Infecções mistas (G1+G2P[4], G2+G3P[4]+P[8], G2+G12P[8]), combinações não usuais (G1P[4], G2P[6]) e cepas atípicas (G3P[3]) também foram identificadas em todo o período do estudo. A vacinação em massa pode alterar o padrão sazonal do RV. A tendência do RV em infectar crianças mais velhas e adolescentes após a implementação da vacina tem sido relatada em todo o mundo. A emergência de G2P[4] segue provavelmente a tendência mundial e, aparentemente, não está relacionada à vacinação. G12 também parece estar emergindo na população brasileira. As rápidas mudanças nos padrões de genótipos dos RVs observados durante o período desse estudo ilustram a existência de uma população dinâmica de cepas selvagens co-circulando no Brasil.

Rotavirus genotypes and the indigenous children of Brazilian midwest in the vaccine era, 2008-2012: Footprints of animal genome.

World group A rotavirus (RVA) surveillance data provides useful estimates of the disease burden, however, indigenous population might require special consideration. The aim of this study was to describe the results of G- and P-types from Brazilian native children ≤ 3 years. Furthermore, selected strains have been analyzed for the VP7, VP6, VP4, and NSP4 encoding genes in order to gain insight into genetic variability of Brazilian strains. A total of 149 samples, collected during 2008-2012, were tested for RVA using ELISA and PAGE, following by RT-PCR and sequencing. RVA infection was detected in 8.7% of samples (13/149). Genotype G2P[4] was detected in 2008 and 2010, G8P[6] in 2009, and G3P[8] in 2011. The phylogenetic analysis of the VP7 and VP4 genes grouped the Brazilian G2P[4] and G3P[8] strains within the lineages currently circulating in humans worldwide. However, the phylogenetic analysis of the VP6 and NSP4 from the Brazilian G2P[4] strains, and the VP7 and NSP4 from the Brazilian G3P[8] strains suggest a distant common ancestor with different animal strains (bovine, caprine, and porcine). The epidemiological and genetic information obtained in the present study is expected to provide an updated understanding of RVA genotypes circulating in the native infant population, and to formulate policies for the use of RVA vaccines in indigenous Brazilian people. Moreover, these results highlight the great diversity of human RVA strains circulating in Brazil, and an in-depth surveillance of human and animal RVA will lead to a better understanding of the complex dynamics of RVA evolution.

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. .

Rotavirus G2P[4] and G2P[4]+[6] infections during norovirus gastroenteritis outbreak: summer season 2010, Brazil.

INTRODUCTION: This study aimed to monitor the seasonality of rotavirus infection, and gain insight into the variability of Brazilian strains. METHODS: A total of 28 stool samples were analyzed from 698 revised cases of gastroenteritis during a norovirus outbreak in the summer of 2010 in Guarujá, Brazil. Diagnosis was performed using enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and sequencing. RESULTS: Rotavirus infection was detected in 17.9% (5/28) of samples; 4 samples were G2P[4] genotype, and one G2P[4]+P[6] genotype. G2 and P[4] sequences showed a genetic relationship to strains from India and Russia, respectively. CONCLUSIONS: The seasonal pattern of rotavirus may be a consequence of human activity apart from climate factors.

Rotavirus G2P[4] and G2P[4]+[6] infections during norovirus gastroenteritis outbreak: summer season 2010, Brazil

Introduction This study aimed to monitor the seasonality of rotavirus infection, and gain insight into the variability of Brazilian strains. Methods A total of 28 stool samples were analyzed from 698 revised cases of gastroenteritis during a norovirus outbreak in the summer of 2010 in Guarujá, Brazil. Diagnosis was performed using enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and sequencing. Results Rotavirus infection was detected in 17.9% (5/28) of samples; 4 samples were G2P[4] genotype, and one G2P[4]+P[6] genotype. G2 and P[4] sequences showed a genetic relationship to strains from India and Russia, respectively. Conclusions The seasonal pattern of rotavirus may be a consequence of human activity apart from climate factors. .

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 animal's 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.