Navegando por Assunto "Genotipagem"

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Acesso aberto (Open Access)
Detecção e genotipagem de norovírus em diferentes amostras de água e esgoto não tratado na cidade de Belém, Pará, Brasil, 2008 a 2010
(Universidade Federal do Pará, 2014) TEIXEIRA, Dielle Monteiro; GABBAY, Yvone Benchimol; http://lattes.cnpq.br/1579859438466504
Enteric viruses excreted in feces from infected individuals dispersed in aquatic environments by sewage discharge. Among these viruses, the norovirus (NoV) is actually considered the main cause of gastroenteritis outbreaks worldwide, resulting from the ingestion of contaminated food and water as well as is also associated with hospitalizations. This research aimed to detect and partially characterize the human NoV (GI/GII) in different water matrices and in untreated sewage from Metropolitan Region of Belem. The study involved superficial waters from bay (Ver-o-Peso), river (Acai’s Port), stream (Tucunduba) and two lakes (Bolonha and Agua Preta), as well as treated water (WTP-Bolonha) and untreated sewage (SLP-UNA), monthly collected over two years . The water and sewage (2 liters) were initially concentrated on filtering membranes to obtain a final volume of 2 mL. The nucleic acid was extracted by silica method and submitted to semi nested RT-PCR (reverse transcription Polymerase chain reaction) using NoV GI and GII specific primers. The cDNA obtained after reverse transcription was also used to investigate the GI/GII by TaqMan® real time PCR. The positive samples for both molecular methods were analyzed for 5’end ORF2 by nested (for GI) and semi nested (for GII) in order to obtain amplicon for identification of circulating strains, being further purified using a commercial kit and submitted to molecular characterization in the automated sequencer. The obtained sequences were edited, aligned and compared to others available in gene bank (NCBI) and in the site NoV genotyping tool. In the period of November 2008 to October 2010, 168 water and sewage samples were collected and analyzed for NoV presence, obtaining a positivity of 33.9% (57/168) of which 21.1% (12/57) were positive only by TaqMan® real time PCR, 19.3% (11/57) only by semi nested and 59.6% (34/57) for both. Considering the two methodologies used, in the positive cases GI (82.5% - 47/57) was most frequent than GII (79.0% - 45/57). However, in most samples there was coexistence of the two genogroups (61.4% - 35/57), mainly in the Tucunduba and SLP-UNA samples, considered the most NoV contaminated sites. On the other hand, in WTP-Bolonha this agent was not found. Of 57 positive samples by TaqMan® real time PCR and/or semi nested RT-PCR, 53 were retested for 5’end ORF2, since four samples showed insufficient quantity of material which allowed a new analyze, so, in 47.2% (25/53) the NoV genome was detected, of these 12% (3/25) belonging to GI, 24% (6/25) to GII and 64% (16/25) for both. The most frequent GI and GII genotypes were GI.8 (n=8) and GII.4 (n=12), respectively, but others genotypes were also observed with lower incidence as GII.6 (n=3), GII.9 (n=2), GII.12 (n=1), GII.14 (n=1), GI.1 (n=1) and GI.4 (n=2). Due to low quality of sequences obtained, eight samples could not be genotyped for GI and three for GII. Of 96 samples with concentration of thermotolerant coliforms above the recommended, 34 (35.4%) were also NoV positive. Increase on conductivity and total dissolved solids was observed in materials from Ver-o-Peso and Tucunduba, as well as the turbidity was notably higher in these places and the Acai’s Port. In the less rainy period (July to November) there was a trend in positivity increasing for NoV, and in the highest rainfall (December to June) a decrease in the incidence of this agent was noted. The results obtained in the present study indicate the circulation of NoV GI and GII in aquatic environments in Belem, revealing the degradation that these water bodies have suffered, as a result of poverty or lack of sanitation in our city, allowing the permanence of pathogens in these ecosystems, along with its effluents.
Acesso aberto (Open Access)
As hepatites B e C na população carcerária feminina do Pará: prevalência, genotipagem e fatores de risco
(Universidade Federal do Pará, 2015) MORAES, Nayana Maria Leal; MARTINS, Luisa Caricio; http://lattes.cnpq.br/1799493244439769
The characteristic of high-risk behavior from the prison public contributes for a high prevalence of diseases transmitted by sexual or parenteral route. Considering the high rates of hepatitis B and C in prisons, this study aims to identify the main risk factors, the prevalence of these diseases and the founded genotypes. This study is an analytical cross-sectional. The sample was composed of 313 inmates from the Female Recovery Center in the State of Pará, wich agreed to participate of this study and were in good physical and mental health. Blood samples were collected and applied a socio-epidemiological questionnaire. The socio-epidemiological analysis showed a predominance of the age group 25-34 years (44.8%), marital status single (55%), incomplete elementary school education (68%) and 1 minimum family wage (65% ). The variables of age and education showed a statistically significant correlation with markers of HBV infection. Risk factors such as cutting and piercing material sharing, tattoo, hospitalization, dental surgery and not condom use showed high frequency. The variable of hospitalization showed statistics association with markers of HBV infection. Serology by ELISA assay showed that 3% were positive for HBsAg, 15% reagents for Anti-HBc, 23% reagents for anti-HBs and 5% for anti-HCV. In the genotyping of samples was found wich from of 10 HBsAg positive samples, 4 samples had undetectable genotype, in 5 samples the genotype E was identified (still not mentioned in Brazil), and in 1 was identified the genotype F (third most prevalent the country) ; from the 17 positive samples for Anti-HCV, 41.2% had undetectable genotype, this same percentage was obtained for genotype 1, and in 17.6% of samples was found the genotype 3, in agreement with the Brazilian standard described in literature.
Acesso aberto (Open Access)
Prevalência e variabilidade genotípica de Chlamydia trachomatis em amostras cervicais de estudantes universitárias em Belém, Pará, Brasil
(Universidade Federal do Pará, 2015) SANTOS, Leonardo Miranda dos; SOUSA, Maisa Silva de; http://lattes.cnpq.br/1775363180781218
Infection with Chlamydia trachomatis is a Sexually Transmitted Infection (STI) more prevalent bacterial in the world and can be asymptomatic in up to 80% of cases and is associated with late complications. The university students are part of a differentiated demand of the population for their high level of education. Objective was to assess the prevalence and variability of the genotypes of C. trachomatis in cervical infection of public university students in the state of Pará, Brazil, and to evaluate the association of the respective socio-behavioral characteristics and gynecological complaints. They included 438 college students from September 2012 to October 2014 and cervical samples were obtained during gynecological examination. It was performed phenol-chloroform technique for extracting total DNA from the sample cervical secretion, and for the detection of C. trachomatis used the Polymerase Chain Reaction (PCR) PCR-seminested the type for amplification 224pb gene omp1. For the identification of genotypes, we carried out a nested PCR for the amplification of the gene omp1 990pb, which was purified and subjected to ABI3130 sequencer, then the nucleotide sequences were compared with GenBank. The prevalence of cervical infection by C. trachomatis was 12.5% (CI: 95% ± 5.89) and genotypes were identified J genotype (36.3%), followed by D genotypes (18.2%) E (18.2%), F (18.2%) and la (9.1%). There was no significant association for age, first sexual intercourse, number of partners, and condoms are used, the presence of gynecological complaints and genotypes found in the study population. Although the prevalence found present itself high among university students, the lack of statistical significance may be due to small sample size and/or result in socially acceptable answers. Efforts are made to trace the expansion of C. trachomatis infection in restricted populations.