RESUMEN
Chronic hepatitis C [CHC] is a major contributor to cirrhosis and hepatocellular carcinoma and major global public health problem that causes mortality in both developed and developing countries. For the past decade, treatment with pegylated interferon [peg interferon alpha] ribavirin [RBV] has been associated with rates of sustained virologic response of = 66% among patients with hepatitis C virus [HCV] infection. In this study, we report the response rate of Iranian treatment-nave CHC patients to Pegaferon, a locally developed pegylated interferon-alpha2a [PEG-IFN alpha2a]. Patients diagnosed with CHC who referred to two university based outpatient clinics in Tehran from December 2007 to May 2011 were enrolled in a single-group, open-labeled experimental design. Eligible patients were above 15 years of age and had HCV infection with evidence of chronic hepatitis. Exclusion criteria included the presence of a debilitating disease, decompensated cirrhosis or refusal to participate in the study. Patients were treated with 180 micro g Pegaferon weekly in addition to 800-1200 mg daily, weight-based RBV for 24 or 48 weeks depending on genotype. Viral response and adverse effects were recorded. A total of 216 patients were enrolled in the study of which 83.3% were male and 16.7% were female. In 93 [43.1%] patients, the HCV RNA viral load was >/= 800,000 IU/ml before starting treatment. "As-treated analysis" indicated that a total of 168 [77.8%] patients achieved sustained viral response [SVR, undetectable plasma HCV RNA 24 weeks after the last planned dose of study treatment]. This study, with a larger number of participants, confirms the results of a previous study by the authors that Pegaferon, a PEG-IFN alpha 2a locally produced in Iran, is effective in treatment-nave CHC patients
RESUMEN
We intend to design and validate a low-cost assay for the detection of hepatitis C virus [HCV] RNA using rapid-cycle RT-PCR. The procedure is performed in a closed system with little risk of contamination allowing PCR and product identification to be performed within one or two hours. A SYBR Green-based real-time RT-PCR for rapid detection of HCV. Amplicon synthesis was monitored continuously by SYBR Green I, which binds to double stranded DMA during PCR. The PCR products were identified by melting curve analysis. Standard sera with known concentrations of HCV RNA and 150 clinical samples were used to validate our assay. The minimum detection level of our assay was less than 50 ID/mL. The results on 100 plasma samples were comparable with commercial assays. This method is useful for rapid qualitative detection of HCV infection and particularly suitable for routine diagnostic applications
Asunto(s)
Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Colorantes Fluorescentes , Hepacivirus/genética , Hepacivirus/aislamiento & purificación , Sensibilidad y Especificidad , Factores de Tiempo , Compuestos Orgánicos , ARN Viral/análisis , ARN Viral/sangreRESUMEN
Hepatitis C virus is the major cause of viral hepatitis and its diagnosis in suspected specimens is of great importance. The risk of transfusion- transmitted virus infection is primarily the result of failure in serological screening tests to detect recently infected donors in the pre-seroconversion window period of infection. Therefore, sensitive and accurate diagnosis of HCV prior to antibody production to reduce window period is necessary. In the present study, a sensitive and specific RT-Nested PCR method for detection of a conserved HCV 5 UTR sequence was developed. Two pairs of primers for amplification of the target sequence in two rounds of PCR were selected. The developed RT- Nested PCR assay was performed on HCV-antibody confirmed positive samples as well as negative controls and standard samples. In order to compare the results, One Step RT-PCR kit was used in this study. 25 HCV-positive plasma samples whose positivity were confirmed by ELISA and Western Blot tests, also as well as 10 fold dilutions of a high viral load plasma sample obtained from a HCV-positive patient as standard samples and 25 negative control plasmas from healthy blood donors were collected and tested by this assay. In all of positive samples a 175bp band was observed on agarose gel electrophoresis, but no band could be detected in negative control plasma. Results from developed RT-PCR assay and One Step RT-PCR kit showed a good correlation. According to the results of this study, the developed RT-Nested PCR assay has a good sensitivity and specificity for diagnosis of HCV infection. It has the advantage of viral genome detection prior to seroconversion and can be used to detect HCV infection during window period