, Emeryville, CA). PCR products of S gene were purified and directly sequenced on ABI PRISM 3100 Genetic analyzer using Big Dye Terminator 3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA). HCV-RNA was extracted from serum samples by Trizol L.S reagent (GIBCO BRL, Life Technologies, Maryland, MD). HCV genotyping was performed Cyclopamine manufacturer
by restriction fragment length polymorphism (RFLP) by previously described method. The results of mixed-genotype infection by RFLP typing were evaluated by direct nucleotide sequencing. Serum HCV-RNA was quantified by a commercially available assay (Amplicore HCV assay; Roche Diagnostic System, Branchburg, NJ). The patients were grouped on the basis of HCV viral load (high and low). A low virus load was defined as less than 100,000 copies/ml by real time (RT)-PCR assay by Amplicore and a high virus load was defined otherwise. HBV-DNA was quantified by TaqMan real-time PCR assay (Applied Biosystems). The HBV buy BI 2536
and HCV sequences were submitted to the comparative sequence program BLAST (www.ncbi.nlm.nih.gov/blast/) for searching other sequences with the highest degree of similarity to identify the genotype. Sequences of isolates were aligned with representative sequences for each major genotype and subtype selected from the GenBank database with the help of the Multalign program. The phylogenetic analysis of HBV and HCV isolates was performed with MEGA 3.0 software. Quantification of AFB1 in food samples by competitive enzyme-linked immunosorbent assay (RIDASCREEN-AFB1, RBIOPHARM, Germany). The number of samples containing aflatoxin had GSK-126
been divided into two groups, viz. those containing less than or equal to 30 ��g/kg (��30 ppb) and above 30 ��g/kg (>30 ppb), keeping in view the tolerance levels recommended by the Codex Alimentarius Commission as well as the regulatory limits of countries such as India. Comparisons between proportions were performed using the ��2 test and exact methods when necessary. Prevalence rates of various etiological factors were calculated to reflect the relative frequency among HCC cases and control. Statistical analyses were performed to compute the odds ratios (ORs) for each risk factor and their 95% CIs by comparing HCC cases with the controls (chronic liver disease patients without HCC), using unconditional logistic regression model for the multivariate analysis based on univariate results. In a subanalysis, a 1:1 pair-matched case�Ccontrol subjects were enrolled. Of 348 HCC cases, 285 were randomly selected to be pair-matched by age and sex (��5 years) to the controls. The conditional logistic regression analysis was performed in this subset of pair-matched by age and sex for multivariate analysis. In our study, the results from conditional logistic regression analysis were consistent with the results from the unconditional logistic regression model. For all tests, p < 0.05 was considered statistically significant. The analysis was performed using SPSS version 12.