The level of disease proceeding are related to different factors, for example, genotype/subtype and viral load. Six important genotypes of HCV and multiplex subtypes have been determined worldwide, HCV Genotyping is considerable clinical option before planning therapy. The proof is that various genotypes of HCV will need diverse term and of course the anti-viral treatment. The purpose of the current study was to assign the dispensation sample of different genotypes of HCV showed in the research reign and their relationship with the viral load in patients suffering suffer from HCV. Patients having positive serology for HCV were chosen for current research (N=105). After that, viral load and genotyping anticipation were carried out utilizing Real-time PCR 59.05 percent (62/105) of samples were contaminated with genotype 3a and consequently genotype 1a and genotype 1b were detected in 13.33percent (14/105) and 8.57percent (9/105) of samples respectively. However, genotype 3 didn’t deal with a prominently higher viral load in comparison with the other genotypes (P= 0.113). Nevertheless, the prominently higher viral load was detectable among male HCV patients (P=0.012) in contrast to women infected with HCV The current research shows that genotype of HCV 3 computed mostly for almost 70.47 percent of the HCV septicity in Isfahan city. Moreover, genotype 3 was not relevance with more extremity of liver disease in contrast to other genotypes as estimated by viral load.