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ABSTRACT: The polymorphism of species A rotavirus genotype G1 strains (RVA-G1) circulating in Rio de Janeiro between 1996 and 2004 was evaluated. The VP7 encoding gene of 36 G1 isolates was sequenced and compared to references strains. The deduced amino acid sequences were used as basis for in silico analysis of the VP7 protein. We observed the circulation of two major G1 lineages and five sublineages during the studied period. Comparison between the VP7 trimeric structures of a rotavirus vaccine strain and Brazilian G1 strains showed mutations at amino acid residues located at the calcium binding site and at several neutralizing antibody recognition sites. Although the rotavirus vaccine program has clearly been successful in Brazil, these results suggest the possibility of the emergence of G1 strains that could evade the immune response elicited by a RVA vaccine and cause a vaccine breakthrough. Consequently, continuous monitoring of rotavirus intragenotypes diversity is critical to understand how it could affect vaccine effectiveness.
Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 05/2012; 12(7):1397-404. · 3.22 Impact Factor
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ABSTRACT: Acute diarrheal disease is still one of the major public health problems worldwide. Rotaviruses (RV) are the most important viral etiologic agents and children under five years of age are the target population.
To investigate the rate of RV infection in hospitalized patients due to acute diarrhea in the cities of Ponta Grossa, Londrina and Assai - Paraná.
Latex agglutination (LA); immunochromatography (ICG); polyacrylamide gel electrophoresis (PAGE) and negative staining electron microscopy (ME) tests were used to detect the virus. For the genotyping, RT-PCR and RT-PCR-ELISA were used, respectively, for NSP4 and VP4/VP7.
Out of 124 samples there were 69 positive stool samples for RV, for at least one of the used tests, 67 of them being RV group A (RV-A). Overall, most of the RV positive stool samples came from children under thirteen years of age. However, 12 positive cases occurred in patients aged 13 years or above, including an 81-year old patient.
The data showed similar electropherotypes and genotypes G, P and NSP4 of the inland wild circulating strains of RV.
The Brazilian journal of infectious diseases: an official publication of the Brazilian Society of Infectious Diseases 12/2010; 14(6):553-7. · 0.55 Impact Factor
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ABSTRACT: A total of 162 rotavirus strains detected between 1996 and 2006 among individuals with diarrhea in Rio de Janeiro, Brazil, were analyzed by multiple-gene genotyping. Characterization of strains was done by RT-PCR assay for amplification and typing of the VP7-, VP4-, VP6-, and NSP4-encoding genes. Overall, 139 (85.8%) strains belonged to the common group A rotavirus combinations: 67 (41.4%) belonged to genotype G1-P[8]-I1-E1; 18 (11.1%) were G2-P[4]-I2-E2; 11 (6.8%) were G3-P[8]-I1-E1; 12 (7.4%) were G4-P[8]-I1-E1; and 31 (19.1%) were G9-P[8]-I1-E1. Two samples presented mixed genotypes (G1 + G3-P[8]-I1-E1 and G1 + G9-P[9]-I1-E1) and rare combinations, such as G2-P[6]-I2-E2 and G9-P[6]-I2-E2, were detected in six (3.7%) strains. The results suggest a linkage among all four genes. Genotypes G1/G3/G4/G5/G9-P[8] were correlated strongly to I1-E1 genotypes and G2-P[4]/P[6] were correlated to I2-E2 genotypes. Unusual combinations of genes, such as G3-P[9]-I2-E2, G9-P[9]-I1-E2, and G3-P[9]-I3-E3, were observed in 15 (9.3%) strains. The characterization of multiple genes allows a more complete analysis of the rotavirus isolates and provides evidence of natural reassortment of strains.
Journal of Medical Virology 10/2010; 82(10):1797-802. · 2.82 Impact Factor
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ABSTRACT: The newly described human bocavirus (HBoV) species 2 and 3 have been repeatedly detected in stool strengthening the possibility that these viruses might present a tropism for the gastrointestinal tract and may be etiological agents of diarrhea.
In this study we assessed the presence of HBoV2 and HBoV3 in stool specimens from Brazilians with acute gastroenteritis.
Stool samples from Brazilian patients with acute diarrhea were analyzed for HBoV2 and HBoV3 by PCR assay. Full or partial genome sequences were obtained for selected isolates. Electron microscopy analysis was used to investigate virus morphology.
Electron microscopy confirmed the presence of virus-like particles in HBoV PCR-positive specimens, with morphology similar to other members of the Parvoviridae family. Five samples out of 807 (0.6%) were positive for HBoV3. Three of the HBoV3-positive patients were HIV/AIDS positive. A selected group of 144 samples was also tested for HBoV2 and 30 samples (20.8%) were positive, 11 of which were HIV/AIDS positive.
This study reports the detection and genetic characterization of HBoV3 and HBoV2 in the stool of Brazilian patients with acute diarrhea. This is the first description of HBoV3 outside Australia, suggesting a wide global distribution of this virus. Further studies are needed to better understand the role of HBoV in gastrointestinal infections, particularly among patients with HIV/AIDS.
Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 04/2010; 48(2):127-30. · 3.12 Impact Factor
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ABSTRACT: Human bocavirus (HBoV) was detected in 14 (2%) of 705 fecal specimens from Brazilian children with gastroenteritis. Coinfection with rotavirus, adenovirus, or norovirus was found in 3 (21.4%) HBoV-positive specimens. None of the HBoV-positive patients had respiratory symptoms.
Emerging infectious diseases 12/2007; 13(11):1756-8. · 6.17 Impact Factor
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ABSTRACT: Crude extract (CE) and aqueous (AqF) and ethyl acetate (EtOAcF) fractions of Guazuma ulmifolia Lam., Sterculiaceae and the corresponding AqF, EtOAcF of Stryphnodendron adstringens (Mart.) Coville, Leguminosae were tested for their antiviral activity against poliovirus 1 (P-1) and bovine herpesvirus 1 (BHV-1) in HEp-2 cultured cells. The antiviral activity was monitored by plaque assay and immunofluorescence assay (IFA) under virucidal and therapeutic protocols. The therapeutic protocol demonstrated statistically significant positive results with both plants and for both virus strains. The highest percentages of viral inhibition were found for G. ulmifolia EtOAcF which inhibited BHV-1 and P-1 replication by 100% and 99%, respectively (p<0.05, Student's t-test). For S. adstringens, AqF was the most efficient, inhibiting BHV-1 and P-1 by 97% and 93%, respectively (p<0.05). In the virucidal protocol, G. ulmifolia CE inhibited the replication of BHV-1 and P-1 by 60% and 26%, respectively (p<0.05), while, for S. adstringens, inhibition of 62% (p<0.05) was demonstrated only with EtOAcF for P-1. IFA demonstrated that the greatest reduction in fluorescent cell number occurred with G. ulmifolia, under the therapeutic protocol for both virus strains. However, AqF and EtOAcF of S. adstringens were most efficient with the virucidal protocol for P-1. In conclusion, we demonstrated that G. ulmifolia and S. adstringens inhibited BHV-1 and P-1 replication, as well as, blocked the synthesis of viral antigens in infected cell cultures.
Biological & Pharmaceutical Bulletin 06/2006; 29(6):1092-5. · 1.66 Impact Factor
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ABSTRACT: The aqueous extract of Agaricus blazei Murill ss. Heinem, a basidiomycete native from Brazil, frequently used by popular medicine, mainly in the form of tea, was assessed to its antiviral action against herpes simplex type 1 (HSV-1) and bovine herpes type 1 (BoHV-1) in HEp-2 cell culture. Viral replication inhibition was evaluated by plaque assay and immunofluorescence test. The extract demonstrated virucide action for both viruses, being more effective against HSV-1, inhibiting its infectivity in 78.4 and 73.9% at the concentrations of 50 and 100 µg/mL, respectively moreover, reduction in 47% the number of fluorescent cells was observed for both concentrations. The extract also showed discrete therapeutic activity. These results suggest that A. blazei extract acts mainly in the viral particle, however, the effect during virus replication can not be ruled out.
Brazilian Journal of Microbiology - BRAZ J MICROBIOL. 01/2006; 37(4).
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ABSTRACT: Acridine orange is a metachromatic intercalator used extensively in histochemistry to differentiate double- from single-stranded (ds, ss) nucleic acid by the emission of green and red fluorescence, respectively, under ultraviolet light. In the present study we standardised a protocol in order to use acridine orange to detect rotavirus ds RNA in polyacrylamide gels and compared it to silver and ethidium bromide staining. We demonstrated that the simplest and best condition was attained when gels containing rotavirus ds RNA bands, stained in green, were treated with 4.3 microM acridine orange after electrophoresis and destained with distilled water pH 6 at 37 degrees C. Under this protocol, rotavirus RNA concentration was calculated and the mean minimum amounts of nucleic acid detected by acridine orange, ethidium bromide, and silver staining were 26.0 +/- 4.29, 15.6 +/- 1.48 and 1.06 +/- 0.11 ng, respectively. The comparison of acridine orange sensitivity with ethidium bromide and silver staining, for 25 field strains of rotavirus and one cell-adapted strain (SA11), demonstrated concurrent results in 80% of the specimens. Red colour emission resulting from the interaction of acridine orange with ss nucleic acid was also shown by testing denatured 0.5 kb HindIII digest of lambda phage DNA. Furthermore, it was demonstrated that rotavirus ds RNA could be used for reverse transcription activity, followed by PCR amplification, after acridine orange staining. In conclusion, although acridine orange is less sensitive than ethidium bromide and silver staining, its practicality, low cost, metachromatic properties, and its non-interference on RT-PCR should be considered. It is suggested the use of acridine orange as an appropriate stain for various purposes in virology, as well as for the molecular biology of nucleic acid.
Journal of Virological Methods 01/2004; 114(1):29-35. · 2.01 Impact Factor
