Search PubMed⌕ Search

Biomedical subjects

H G Pereira

Publications and source records attributed to H G Pereira.

At least 19 recordsLinked to original sources
PubMed1993

Enteric viruses and diarrhea in HIV-infected patients. Enteric Opportunistic Infections Working Group.

BACKGROUND: Diarrhea occurs frequently among persons with the acquired immunodeficiency syndrome, but the cause often remains unknown. We used a group of diagnostic assays to determine which viruses were etiologic agents of diarrhea in a group of persons infected with the human immunodeficiency virus (HIV). METHODS: Stool and serum specimens were obtained from HIV-infected patients enrolled in a longitudinal study in Atlanta. Fecal specimens from patients with diarrhea and from control patients without diarrhea were screened by electron microscopy, polyacrylamide-gel electrophoresis, and enzyme immunoassays for rotaviruses, enteric adenoviruses, caliciviruses, picobirnaviruses, and astroviruses. Paired serum samples were tested for antibody responses to Norwalk virus and picobirnavirus. RESULTS: Viruses were detected in 35 percent of 109 fecal specimens from patients with diarrhea but in only 12 percent of 113 specimens from those without diarrhea (P < 0.001). Specimens from patients with diarrhea were more likely than those from patients without diarrhea to have astrovirus (12 percent vs. 2 percent, P = 0.003); picobirnavirus (9 percent vs. 2 percent, P = 0.017); caliciviruses, including small round structured viruses (6 percent vs. 1 percent, P = 0.062); and adenoviruses (9 percent vs. 3 percent, P = 0.047). They were also more likely to have a mixed viral infection (6 percent vs. 0 percent, P = 0.006). With the use of polyacrylamide-gel electrophoresis to analyze concentrated RNA extracts from stool, picobirnavirus was detected in fecal specimens from 6 of the 65 patients with diarrhea and was associated with prolonged viral shedding and chronic diarrhea. No rotaviruses, enteric adenoviruses, or instances of seroconversion to positivity for Norwalk virus were observed. CONCLUSIONS: Novel enteric viruses such as astrovirus and picobirnavirus may be more important etiologic agents of diarrhea in HIV-infected patients than previously recognized and may be more common than either bacterial or parasitic enteropathogens.

Adenoviruses, Human↗
PubMed1993

National laboratory surveillance of viral agents of gastroenteritis in Brazil.

In 1984 an enzyme immunoassay for rotavirus and adenovirus developed in the Virology Department of the Oswaldo Cruz Foundation was distributed to laboratories in 14 Brazilian states as part of a project to survey viral agents in fecal specimens from children with diarrhea. The ensuing surveillance continued for 3 to 5 years, and in 1991 the results obtained by all laboratories that tested 300 or more fecal specimens were reviewed at a workshop held in Belém. These results showed that rotavirus had been detected in 13% to 20% of the specimens tested, with positive specimens appearing to peak in the May to September "winter" or dry season in Brazil's central and southern states but not in the more tropical northern areas. Adenovirus, detected in 0.7% to 5.5% of the specimens tested for it, showed no seasonal variations. Many other viral agents known to cause gastroenteritis (e.g., astrovirus, small round-structured viruses, calcivirus, and group C rotavirus) were detected at centers that used electron microscopy. Picobirnavirus, a novel agent not yet associated with gastroenteritis in humans, was found by polyacrylamide gel electrophoresis in specimens from a number of centers using this technique. Vaccines to prevent rotavirus that are currently under development would be of great use in Brazil, where rotavirus is the most common cause of childhood diarrhea. Improved diagnostics will be required to assess the importance of the other viral agents.

Acute Disease↗
PubMed1990

A novel avian virus with trisegmented double-stranded RNA and further observations on previously described similar viruses with bisegmented genome.

The occurrence in chickens of small viruses with bisegmented double-stranded RNA (dsRNA) genome is confirmed and a new virus with similar properties but with three genome segments is described. Both differ from birnaviruses (Intervirology 25, 141-143, 1986) in having indistinct surface structure, smaller diameters (35 nm), and higher buoyant density (1.4 g/ml) in CsCl but are similar in these respects to viruses previously described in several mammals (Lancet 2, 103-104, 1988; J. Gen. Virol. 69, 2749-2754, 1988; Res. Vet. Sci, in press) under the tentative name of picobirnaviruses (PBV). Genome segment length estimations gave values of 2.6 and 1.9 kbp for the avian PBV and 2.9, 2.4 and 0.9 kbp for the trisegmented viruses. The source and pathogenic potential of these viruses remain to be established.

Animals↗
PubMed1989

New genome types of adenovirus types 1, 3, and 5 isolated from stools of children in Brazil.

During an epidemiological survey made in São Paulo (Brazil), fecal specimens were periodically collected from 100 randomly chosen babies from birth to the age of 18 months. The stools, routinely collected each month and also collected each time a child presented any sign of disease, were screened for the presence of adenoviruses. Sixteen adenovirus strains, isolated from the stools of healthy and ill children, were characterized by restriction enzyme analysis. Five isolates were from subgenus A, five were from subgenus B, four were from subgenus C, and two were from subgenus D. All but two showed some restriction patterns different from those of the 42 human adenovirus prototypes and all the genome types described up to now. No fastidious adenovirus (subgenus F, serotypes 40 and 41) was encountered in the stools examined. We report here the restriction enzyme analysis of isolates of subgenera B and C. The following new designation genome types are proposed: Ad3e1 (subgenus B) and Ad1d, Ad5a1, and Ad5a2 (subgenus C).

Adenoviridae Infections↗
PubMed1989

Viruses with bisegmented double-stranded RNA in pig faeces.

Viruses similar to the bisegmented double-stranded (ds) RNA picobirnaviruses described in human faeces and the intestinal contents of Oryzomys nigripes rats and guinea pigs were isolated from the faeces of pigs taken from several areas in the state of Sao Paulo, Brazil. Samples were collected from 912 pigs of several breeds, aged nine to 61 days, and assayed by polyacrylamide gel electrophoresis with silver staining and a combined enzyme immunoassay for rotavirus and adenovirus, using the simian rotavirus SA11 as control. Electrophoretic profiles resembling the bisegmented dsRNA viruses were detected in 106 pigs with 15.3 per cent occurring in animals with diarrhoea compared to 9.6 per cent in animals without diarrhoea.

Animals↗
PubMed1988

A virus with a bisegmented double-stranded RNA genome in rat (Oryzomys nigripes) intestines.

Examination of the intestinal contents of free-living Oryzomys nigripes rats by PAGE revealed two sharply defined bands that could be stained by ethidium bromide or by silver nitrate with comparable intensities. The molecules forming these bands were susceptible to digestion by pancreatic RNase A but not by RNase T1 or by DNase I. Their lengths were estimated to be about 2.6 and 1.5 kbp, respectively, by comparison with rotavirus SA11 genome segments. They cosedimented in CsCl gradients at a density of 1.39 to 1.40 g/ml, together with uniform particles approximately 35 nm in diameter with indistinct surface structure. It is suggested that these particles represent an as yet undescribed virus with a bisegmented double-stranded RNA genome, for which the name 'picobirnavirus' is proposed.

Animals↗
PubMed1987

Dot enzyme immunoassay. A simple, cheap and stable test for antibody to human immunodeficiency virus (HIV).

A dot enzyme immunoassay for antibody to HIV has been developed and tested with a panel of positive and negative sera. It has proved to be of equal or greater sensitivity compared with a commercial ELISA kit, is simple and quick to perform, requires neither sophisticated equipment nor highly trained technical staff. The reagents are stable enough for postal distribution in tropical countries and, other than for the antigen, the costs are low, making it an appropriate test for use in the developing world when funds for expensive commercial kits are not available.

Acquired Immunodeficiency Syndrome↗
PubMed1987

A combined dot nitrocellulose-enzyme immunoassay for rotavirus and adenovirus.

A combined enzyme immunoassay using nitrocellulose membrane as solid support is described for the detection of rotavirus and adenovirus in faeces from children with gastroenteritis. Its sensitivity and specificity are comparable to those of a previously described assay performed on plastic microplates (Pereira et al. (1985) J. Virol. Methods 10, 21-28). The introduction of nitrocellulose membrane as support for the immune reactions greatly simplifies the multiple washing steps and precludes the need for disposable plastic plates.

Adenovirus Infections, Human↗
PubMed1986

Diarrhoea in mice infected with a human rotavirus.

Oral inoculation of newborn mice with the MET strain of human rotavirus produced transient diarrhoeal disease. Light and scanning electron microscopy showed typical rotavirus-induced morphological lesions in the villous epithelium of the small intestine consisting of extensive cytoplasmic vacuolation, villous necrosis and atrophy. Virus recovered from intestinal suspensions of infected mice showed the typical electrophoretic profile of the genome of the inoculated strain. Rotavirus antibody appeared in infected mice 10 to 20 days after inoculation but not in controls or nursing dams. The availability of a small animal model for experimental infection with human rotaviruses should prove useful for virulence and protection studies.

Animals↗
PubMed1986

Direct detection and differentiation of fastidious and nonfastidious adenoviruses in stools by using a specific nonradioactive probe.

Restriction fragments of fastidious human adenovirus type 41 (Ad41) were cloned in vector plasmid pBR322. A rapid and sensitive nonradioactive molecular-hybridization technique (M. Renz and C. Kurz, Nucleic Acids Res. 12:3435-3444, 1984) showed that one clone specifically detected fastidious Ad40 and Ad41 (subgenus F) without cross-hybridization with nonfastidious adenoviruses. This clone was mapped in a region of the Ad41 genome corresponding to early transcription unit E1B of Ad2. A number of DNAs from fastidious and nonfastidious adenoviruses were extracted, without cultivation, from stools of children with gastroenteritis and were hybridized with an Ad2 probe and with the cloned probe, allowing the differentiation of the two groups of viruses. This method could detect DNA quantities as low as 10 pg and should be particularly suitable for stool samples containing adenoviral DNA in amounts too low to be detected by staining with ethidium bromide.

Adenovirus Infections, Human↗
PubMed1986

Electrophoretic study of the genome of human rotavirus from Maceió, Brazil.

Rotaviruses were detected by enzyme immunoassay (EIA) in 53 (13.3%) of 397 fecal samples from children with acute gastroenteritis in the city of Maceió, Alagoas, Brazil. Polyacrylamide gel electrophoretic (PAGE) patterns characteristic of rotavirus double-stranded RNA were detected in 51 (96.2%) of the 53 EIA-positive samples. Of the RNA-positive samples, 1 (2%) was classified as subgroup 1 (short profile), 49 (96%) as subgroup 2 (long profile) and 1 (2%) could not be classified because of the absence of bands 10 and 11. The strains of subgroup 2 showed a great degree of electrophoretic heterogeneity and could be divided into several subcategories. Two samples showed splitting of one of the genome segments. PAGE, a very sensitive method capable of identifying rotavirus RNA genomes, has demonstrated that human rotaviruses detected in Maceió present many differences in RNA electrophoretic patterns.

Acute Disease↗
PubMed1985

Adenoviruses in faeces of children with acute gastroenteritis in Rio de Janeiro, Brazil.

Faeces from 746 children less than 5 years old with acute gastroenteritis were screened for the presence of adenovirus particles or antigens by immunoelectron microscopy (IEM) and enzyme immunoassay (EIA). Thirty-five samples were positive by both IEM and EIA, two only by IEM, and two only by EIA, giving a total of 39 (5.2%) samples with positive results. Of these, 25 could be propagated in HEp2 cells and were neutralized by one of the antisera to adenovirus types 1 to 18. The remaining 14 samples could be propagated only in the 293 permanent line of human cells transformed by adenovirus type 5 DNA [Graham et al, 1977] and were not neutralized by antisera to adenovirus types 1 to 31. An EIA carried out by the antibody-capture technique, using antiserum specific for "enteric" adenoviruses [Johansson et al, 1979], gave positive results with all isolates that could be propagated only in 293 cells and with none of those capable of growing in HEp2 cells.

Adenoviruses, Human↗
PubMed1985

In situ hybridization with biotinylated DNA probes: a rapid diagnostic test for adenovirus upper respiratory infections.

Adenovirus DNA was detected in cells from nasopharyngeal secretions of children with acute respiratory infections by in situ hybridization with biotinylated probes. The technique was easy to perform, giving rapid results which were well correlated with those of immunofluorescence assays of the same samples. Adenoviruses of subgroups B, C and E were detected equally well by probes prepared either from purified adenovirus type 5 or from a plasmid (A1) carrying a cloned insertion of BamH1 fragments C and D of adenovirus type 2 in pAT 153. The use of stable non-radioactive probes makes in situ hybridization a feasible assay for use in clinical laboratories with moderate resources.

Adenoviridae Infections↗
PubMed1985

A combined enzyme immunoassay for rotavirus and adenovirus (EIARA).

A combined enzyme immunoassay for rotavirus and adenovirus (EIARA) was developed as a double-antibody sandwich assay in which test samples are added to plastic wells coated with rotavirus or adenovirus goat antibody. The presence of antigens is detected by mixed-guinea pig antisera to the same viruses followed by rabbit anti-guinea pig IgG conjugated with peroxidase and subsequently by ortho-phenylenediamine substrate. Titrations of rotavirus (SA11) and adenovirus type 2 tissue culture grown viruses in the presence of separate capture sera and mixed detector sera revealed that the two virus-antibody reactions occurred independently and were not affected by each other. Comparison with another enzyme immunoassay for rotavirus, with immunoelectron microscopy and with polyacrylamide gel electrophoresis showed that EIARA is a sensitive and specific method for detecting rotaviruses and adenoviruses in faeces from children with gastroenteritis.

Adenoviruses, Human↗
PubMed1984

Genomic heterogeneity of simian rotavirus SA11.

A preparation of simian rotavirus SA11 was shown to contain, in addition to the normal 11 genome segments, an RNA species with electrophoretic mobility slightly higher than that of segment 4. Limiting dilution passages allowed the separation of two virus clones distinguishable from each other by the electrophoretic mobility of that genome segment. Possible implications of this finding in virus behaviour and in the comparison of rotaviruses by RNA electrophoresis are discussed.

Animals↗