SARS-CoV-2 RNA detection in stool samples from acute gastroenteritis cases, Brazil.

We described the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in stool samples from patients presenting only acute gastroenteritis (AGE) symptoms. From January to July 2020, 121 AGE stool samples were screened by quantitative reverse-transcription polymerase chain reaction. We detected SARS-CoV-2 in 27.5% of samples received during the epidemic period. No infectious viruses were observed in Vero E6 cells.

The evolving epidemiology of rotavirus A infection in Brazil a decade after the introduction of universal vaccination with Rotarix®.

BACKGROUND: Brazil introduced the monovalent rotavirus vaccine (Rotarix®) in 2006. This study aimed to assess the epidemiology and genotype distribution of species-A rotavirus (RVA) in Brazil, comparing the pre- and post-vaccination periods. METHODS: Laboratory-based RVA surveillance included 866 municipalities in 22 Brazilian states, over a 21-year period. A total of 16,185 children with diarrheal diseases (DD) aged up to 12 years between 1996 and 2005 (pre-vaccination period, n = 7030) and from 2006 to 2017 (post-vaccination period, n = 9155) were enrolled. RVA was detected using ELISA immune assay and/or polyacrylamide gel electrophoresis and genotyped using nested PCR and/or nucleotide sequencing. RVA-positivity and genotypes detection rates were compared in distinct periods and age groups and Rotarix vaccination status. RESULTS: RVA-positivity in pre- and post-vaccination periods was, respectively: 4-11 months bracket, 33.3% (668/2006) and 16.3% (415/2547) (p <  0.001); 12-24 months, 28.2% (607/2154) and 22.2% (680/3068) (p <  0.001); 25-48 months, 17.4% (215/1235) and 29.4% (505/1720) (p <  0.001). Genotypes distribution in the pre- and post-vaccination periods was, respectively: G1P [8]/G1P[Not Typed], 417/855 (48.8%) and 118/1835 (6.4%) (p <  0.001); G2P [4]/G2P[NT], 47/855 (5.5%) and 838/1835 (45.7%) (p <  0.001); G3P [8]/G3P[NT], 55/855 (6.4%) and 253/1835 (13.8%) (p <  0.001); G9P [8]/G9P[NT], 238/855 (27.8%) and 152/1835 (8.3%) (p <  0.001); G12P [8]/G129P[NT], 0/871 (0%) and 249/1835(13.6%) (p <  0.001). Concerning infants aged 4-11 months, RVA frequency in fully vaccinated and non-vaccinated individuals was 11.9% (125/1052) and 24.5% (58/237) (p <  0.001), respectively. In children aged 12-24 months, RVA detection rate was 18.1% (253/1395) and 29.6% (77/260) (p <  0.001), for the vaccinated and non-vaccinated individuals, respectively (p <  0.001). CONCLUSIONS: RVA infection was significantly less frequent in children aged ≤2 years with DD after implementing vaccination, mainly among vaccinated children. It was also observed a decrease of P [8] circulation and emergence of G2P[4] in 2005, and afterwards in the post-vaccine era, with spreading of G12P[8] in 2014-2015 and of G3P[8] in 2017. Continuous RVA surveillance must be carried out in this scenario.

Adenoviruses associated with acute gastroenteritis in hospitalized and community children up to 5 years old in Rio de Janeiro and Salvador, Brazil.

Acute gastroenteritis is a major source of morbidity and mortality among young children in developed and developing countries. Human adenoviruses (HAdVs), and in particular species F, are related to childhood diarrhoea worldwide. This study presents the results obtained during an investigation of HAdVs causing acute gastroenteritis in children hospitalized in Rio de Janeiro, RJ, Brazil, from April 1996 to September 2003, as well as in children with diarrhoea living in the slums of Salvador, BA, Brazil, from October 2001 to September 2003. A total of 3060 stool samples was analysed by an enzyme immunoassay for rotavirus and adenovirus (EIARA) and 61 (2%) were found to be positive. HAdV presented with low prevalence throughout the year, with a slight but not significant increase in incidence in late summer and early autumn. Children up to 2 years of age were the most frequently affected (79% of all positive samples). All positive samples were analysed further by generic and species-specific HAdV PCR protocols, confirming 100% specificity of this rapid and inexpensive EIARA. Species F was the most prevalent (65%), despite the occurrence of species A (12%), C, D and co-infection F/D (5% each) and species B and co-infections F/A, F/C and B/D (2% each). In order to type the species F strains as HAdV-40 or -41, generic PCR and a HinfI restriction digest were performed. HAdV-40 and -41 were found to represent 62% (23/37) and 38% (14/37), respectively. These results demonstrated that a combination of generic and species-specific PCRs is useful and reliable for HAdV species and type identification directly from faecal specimens. The results confirmed the endemism of human adenoviruses, mainly species F, in children as aetiological agents of diarrhoea, although the limited sensitivity of EIARA as a screening method may have underestimated their prevalence.

Performance of a one-step quantitative duplex RT-PCR for detection of rotavirus A and noroviruses GII during two periods of high viral circulation.

Rotavirus A (RVA) and noroviruses (NoV) are the major viral agents of acute gastroenteritis (AGE) worldwide. In the present study, we aimed to evaluate the performance of a one-step duplex quantitative RT-PCR (dRT-qPCR) assay, established for detection and quantification of RVA and NoV genogroup II (GII) using a single DNA standard curve (SC), as well as to investigate the association between fecal viral load and optical density (OD) values, and viruses' genotyping. The results obtained by dRT-qPCR in 530 fecal samples from AGE cases were compared with methods employed for the diagnosis of those viruses as follows: enzyme immunoassay (EIA) and polyacrylamide gel electrophoresis (PAGE) for RVA; and qualitative PCR for NoV. By using dRT-qPCR, we detected RVA and NoV in 353 (66%), increasing the positivity rate by 22.5% for RVA and 11.5% NoV, comparing the number of positive samples. RVA and NoV GII were detected in a range of 5.17 × 10(3) to 6.56 × 10(9) and 3.76 × 10(3) to 9.13 × 10(10) genome copies per gram of feces, respectively. We observed a significant direct correlation between genome copies values and optical density, using dRT-qPCR and EIA assays, respectively (Spearman ρ=0.41; p<0.0001). Viruses characterization demonstrated a predominance of NoV GII.4 Sidney 2012 variant during October 2013 to February 2014, followed by the emergence of RVA genotype G12P[8] in 2014. The established assay using a single SC provides an early feedback concerning detection and quantification, with the advantage of detecting simultaneously RVA and NoV GII, reducing time and reagent costs.

G26P[19] rotavirus A strain causing acute gastroenteritis in the American continent

In Brazil, the rotavirus A genotype G26 was first identified in suckling piglets, while the P[19] genotype has not been identified in any animal species so far. This report details the genetic characterisation of a G26P[19] RVA strain detected from an eight year-old child, vaccinated with Rotarix®, hospitalised with acute diarrhoeal disease in Rio de Janeiro in 2015. Most likely, the genome constellation (I5-R1-C1-M1-A8-N1-T1-E1-H1) observed in the G26P[19] Brazilian strain was a result of interspecies transmission events between humans and pigs. In addition, a rearrangement in the NSP5 gene was observed downstream of the 3' non-coding region.

Neonatal rotavirus infection in Belém, northern Brazil: nosocomial transmission of a P[6] G2 strain.

A total of 614 fecal specimens were obtained during a survey for rotavirus infection conducted between May 1996 and May 1998 among 437 newborns admitted to special care nurseries at a public hospital in the urban area of Belém, Brazil. Routine stool samples were taken weekly from all babies up to the age of 28 days. Overall, 51 (11.7%) of the neonates excreted rotaviruses while in hospital, of whom 42 (82.3%) developed asymptomatic nosocomial infection; nosocomial infection was also proved in five of the nine patients with diarrhea. Three distinct RNA profiles were detected, of which one short electropherotyping pattern was far more frequent ( approximately 90% of the strains). Using monoclonal antibody-based enzyme immunoassays, 32 (62.7%) of the rotavirus-positive strains were classified as G2, and 1 (1.9%) as mixed G1 and G2. A G serotype could not be assigned to 18 (35.3%) of the isolates. A reverse transcription-polymerase chain reaction was used for determining the VP4 type-specificity of a subset of 28 rotavirus-positive samples. Characterization of the VP7-genotype specificity was also sought for 18 of these latter strains. Overall, P[6] and G2 genotypes were identified in 93% and 94% of tested samples respectively, with results being further confirmed by Southern hybridization. Although surveillance was conducted during a 25-month period, 50 (98%) of 51 rotavirus isolates clustered between January and December 1997. The earliest [P6]G2 rotavirus infections were detected by late January 1997, involving two (13- and 14-day-old) babies admitted with acute diarrhea. Thereafter, strains bearing these genotype specificities were identified among five infants with hospital-acquired gastroenteritis, followed by 16 others who were infected asymptomatically. This is the first report from Brazil describing nosocomial transmission of P[6]G2 rotavirus strains among neonates.

Rotavirus strain diversity in Rio de Janeiro, Brazil: characterization of VP4 and VP7 genotypes in hospitalized children.

Rotavirus strains from 91 patients treated at a children's hospital from 1996 to 1998 in Rio de Janeiro, Brazil, were characterized by electropherotyping, reverse transcription-PCR amplification for P and G genotypes, and Southern hybridization. Results obtained showed that following predominant [P],G type combination: P[4], G2 (21 per cent), P[8], G1 (17 per cent), P[8], G3 (13 per cent), which are prevalent throughout the world. However, an unexpected number of cases were associated with uncommon genotypes: P[8], G2 (13 per cent), P[8], G5 (11 per cent), P[8], G9 (7 per cent), P[8], G10 (4 per cent), P[6], G4 (3 per cent), P[6], G3 (1 per cent), P[4], G9 (1 per cent), and P[6], G9 (1 per cent). Mixed infections with more than one type were identified in only two cases and 16 per cent of the samples were not G and/or P typeable. A subset of G types was confirmed by Southern hybridization and chemiluminescent detection. Rotavirus seasonal distribution was observed between April and July. The contribution of the results obtained in the present investigation corroborates the required epidemiological surveillance for rotavirus infection in Brazil.