Human papillomavirus (HPV) infection is common in sexually active women and viral persistence may cause intraepithelial lesions and eventually progress to cervical cancer (CC). The present study aimed to investigate epidemiological factors related to HPV infection and to evaluate viral persistence and CC precursor lesions frequencies in women from a city in the countryside of South Brazil. Three hundred women were recruited from a primary public health care clinic. The patients were interviewed and underwent sampling with cervical brushes for HPV-DNA detection/typing by a PCR-based assay and cytological analysis by Pap smear test. HPV was detected in 47 (15.7%) women. HPV infection was significantly associated with young age (<30 years) and low socio-economic status. Seventeen (5.7%) women presented cytological abnormalities, three of them with precursor CC intraepithelial lesions. A subgroup of 79 women had been previously analyzed and thirteen (16.4%) were persistently infected, two with precursor CC intraepithelial lesions and high-risk HPV types infection (both of them without cervical abnormalities in the first exam). In conclusion, HPV infection was associated with young age (<30 years) and low family income; viral persistence was low (16.4%) but related to CC precursor lesions; and HPV-DNA high risk types detection would help to screen CC in the population.
BACKGROUND Hepatitis B virus (HBV) infection is a major public health problem in Brazil. Several risk factors are involved in HBV infection and their identification by a rational and essential approach is required to prevent the transmission of this infection in Brazil.OBJECTIVES To evaluate risk factors associated with HBV infection in South Brazil.METHODS A total of 260 patients with HBV and 260 controls from Caxias do Sul (state of Rio Grande do Sul, Brazil) participated in this study. All participants were given a standard questionnaire to yield the sociodemographic information and to identify HBV risk factors. HBV infection was detected by HBsAg test in all participants.FINDINGSHBV infection in these cases was strongly associated with history of a family member HBV-infected, mainly mother [odds ratio (OR) = 4.86; 95% confidence intervals (CI): 1.69–13.91], father (OR = 5.28; 95% CI: 1.58–17.71), and/or siblings (OR = 22.16; 95% CI: 9.39–52.25); sharing personal objects (OR = 1.40; 95% CI: 1.37–2.38); and having history of blood transfusion (OR = 2.05; 95% CI: 1.10–2.84).CONCLUSIONS HBV infection was strongly associated with having a family member infected with hepatitis B, sharing personal objects, and having history of blood transfusion.
The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.
Introduction: Molecular biology procedures to detect, genotype and quantify hepatitis C virus (HCV) RNA in clinical samples have been extensively described. Routine commercial methods for each specifi c purpose (detection, quantifi cation and genotyping) are also available, all of which are typically based on polymerase chain reaction (PCR) targeting the HCV 5' untranslated region (5'UTR). This study was performed to develop and validate a complete serial laboratory assay that combines real-time nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) techniques for the complete molecular analysis of HCV (detection, genotyping and viral load) in clinical samples. Methods: Published HCV sequences were compared to select specifi c primers, probe and restriction enzyme sites. An original real-time nested RT-PCR-RFLP assay was then developed and validated to detect, genotype and quantify HCV in plasma samples. Results: The real-time nested RT-PCR data were linear and reproducible for HCV analysis in clinical samples. High correlations (> 0.97) were observed between samples with different viral loads and the corresponding read cycle (Ct -Cycle threshold), and this part of the assay had a wide dynamic range of analysis. Additionally, HCV genotypes 1, 2 and 3 were successfully distinguished using the RFLP method. Conclusions: A complete serial molecular assay was developed and validated for HCV detection, quantifi cation and genotyping.
BACKGROUND: It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner. OBJECTIVES: Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene. METHODS: Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay. RESULTS: The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5% increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay. CONCLUSION: The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing
Cervical cancer (CC) is caused by persistent human papillomavirus (HPV) infection and affects women worldwide. The progression of an HPV persistent infection to CC is influenced by genetic factors. Three single nucleotide polymorphisms (SNPs) in TP53, NQO1 and RPS19 genes (rs1042522, rs1800566, rs2305809, respectively) were previously associated with CC in European and North American populations. The present case-control study aimed to investigate the association of the SNPs rs1042522, rs1800566, and rs2305809 with CC in an admixed population in southern Brazil. A total of 435 women (106 CC patients and 329 controls) were recruited for this study. All women were interviewed and underwent clinical sampling. SNPs rs1042522 and rs1800566 were evaluated by PCR-RFLP. SNP rs2305809 was determined by real-time PCR. The crude and adjusted ORs with 95% CI were estimated. The recessive genetic model (C/C + C/T) for rs2305809 was more frequent in the control group (79.9%) compared to the cases (69.8%), being associated with CC protection (adjustedOR = 0.49; 95% CI: 0.27–0.90). However, the other polymorphisms evaluated did not present significant differences between cases and controls. This study detected a protective association for the recessive genetic model in rs2305809. These results suggest a potential role of the RPS19 gene in CC.
Resumo Objetivo Investigar a presença de infecção cervical pelo papilomavírus humano (HPV, human papillomavirus) em mulheres idosas e fatores relacionados. Método Trata-se de um estudo transversal, retrospectivo e descritivo, com abordagem quantitativa. A amostra foi constituída por 106 mulheres com idade igual ou superior a 60 anos atendidas em serviços de saúde pública de uma cidade do Sul do Brasil, as quais realizaram coleta de material cervical para análise citológica e detecção molecular do DNA do HPV, bem como levantamento de dados clínicos e sociodemográficos por meio de um questionário padronizado e requisição do exame citopatológico. Resultados A idade das pacientes variou entre 60 e 82 anos, com média de 64,9 ± 5,1. O HPV foi detectado em 14 (13,2%) idosas avaliadas no estudo e 8 tipos virais foram identificados, a maioria (n=7; 87,5%) de alto risco oncogênico. Observou-se, por análise de qui-quadrado, que casos HPV positivos possuem associação com maior número de parceiros sexuais (p= 0,018). Na citologia, a maioria das mulheres (n=102; 96,2%) apresentou resultado negativo para lesão intraepitelial ou malignidade e duas (1,8%) apresentaram citologia alterada, mas destas, nenhuma apresentou infecção por HPV no teste molecular. Das 10 mulheres avaliadas em duas visitas, sete (70%) não apresentaram infecção pelo HPV em ambas as avaliações, duas (20%) eliminaram a infecção pelo HPV e uma (10%) apresentou conversão para positividade. Nenhuma delas apresentou infecção persistente. Conclusão Mulheres idosas estão suscetíveis à infecção por HPV e às lesões causadas por esse vírus, por isso devem manter o rastreamento citológico.
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