Determinação do RNA-VHC no sêmen de pacientes cronicamente infectados pelo vírus da Hepatite C / Determinations of the RNA-HCV in semen from chronically patients infected by the hepatitis C vírus

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

Introduction: Hepatitis C vírus is a huge problem for public health, and its global prevalence is estimated around 3%. Its transmission by seminal fluid is still in discussion in several fields, such as assisted reproduction and in studies about risk factors, whether the hepatitis C virus is an STD (sexually transmitted disease) or not. Twenty-three patients were investigated. Objectives: 1.Establish a technique to detect the presence or absence of the HCV in semen from chronically infected patients; 2.Compare semen samples handling techniques, in order to decrease the amount of inhibitors on the samples; 3.Compare different PCR and detection techniques for the HCV in semen samples, in order to increase the sensibility of the test. Methods: On the first phase 20 patients were selected (13 filled the inclusion criterion). Semen and serum samples were collected. The semen samples were processed with the help of Percoll® 90% and 45%. The presence of the RNA-HCV were analyzed in serum with Amplicor Roche method, qualitative test. When positive, the serum samples were genotyped and the semen samples were extracted, by the same method, and the PCR was done. On the second phase 23 patients were selected, some of them were old patients from the first phase (20 filled the inclusion criterion). Semen and serum samples were collected. The semen samples were processed through a dilution series. The presence of HCV-RNA was analised by Amplicor Roche, qualitative test and by PCR in Real-time. The epidemiological data and genotypes were analised. Resultados: From the 23 patients selected the mean age was 40,7 years, mean 45 years. The mean time of Discovery was 7,15 years. Ten patients (37,1%) didn´t present any apparent epidemiology, eight patients (29,6%) contracted HCV through injection and inhalatory drug use; six patients (22,2%) through blood transfusion; two patients (7,4%) had history of drug use and blood transfusion and one patient (3,7%) who was a health professional. Genotype 3a was found in 40,7% of the patients, followed by 1a with 26% of the patients, 1b with 14,8%, 2b with 11,1% e 1a/1b in 7,4% of the patients. The samples processed with Percoll, 86,5% presented inhibited results. Whereas on the samples that were processed with dilution series and amplified on the conventional PCR only 25,62% presented inhibited results, 65% were undetected and 9,38% were positive. On the samples processed with dilution series on the Real-time PCR 95% were undetected and only 5% were positive. Conclusion: On the attempt of decreasing the amount of inhibitors found on the semen samples, the procedure of dilution series showed us more efficient results when compared to the Percoll procedure. However, the great amount of undetected showed that the viral load might have being diluted, leading us to the necessity of a more sensitive technique. There was no significant difference between the results of the conventional PCR and the Real-time. These increase on the undetected results may be a consequence of the absence of a internal control on the PCR reactions.

ASSUNTO(S)

vírus da hepatite c/diagnóstico hcv reação em cadeia pela polimerase hcv hepatitis c virus/diagnostic hepatite c/virologia polimerase chain reaction hepatitis c/virology

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