Introduction Blood transfusion is a specialized modality of patient management to sustain health and life, but carries a definite risk of acquisition of transfusion transmitted virus (TTV) infections particularly in resource limited settings. TTV infections are of great concern because of their asymptomatic nature, protracted viremia, highly variable course and of fatal, chronic and life threating disorders. The prevalence of TTVs among the blood recipients reflects the disease burden among the blood donors and general population. Material and method The study population was categorized in two groups on the basis of the history of blood transfusion: individuals with no history of blood transfusion were categorized as control group and individuals with history of blood transfusion were categorized as high risk group. High risk group was further divided into four sub groups; thalassemia, hemophilia, hemodialysis, and surgical patients. Sera of all individuals were investigated for the presence of HBsAg, anti-HCV and anti-HIV by using immune-chromatigraphic tests, followed by molecular analysis by PCR for the detection of viral DNA and RNA and genotypes. Result The study population consists of 4,607 individuals, 2,032 individuals comprised control group and 2,575 individuals comprised high risk group. Overall, 21.12% (973) individuals showed the evidence of TTV, among them 3.6% (166) was positive for HBsAg, 17.39% (801) was positive for anti-HCV and 0.13% (6) was positive to HBV&HCV while no individual was positive to HIV. 75 individuals were positive to HBV DNA, 18 individuals (27.69%) were classified into genotype B, 11 (16.92%) genotype C, 35 (53.84%) genotype D and 1 (1.54%) genotype F, 10 xi (15.38%) untypeable. 366 individuals were positive to HCV RNA, HCV genotypes 1 was detected in 71 (20.9%) individuals, genotype 2 in 17 (5.01%), genotype 3 in 182 (53.69%), genotype 4 in 16 (4.72%), genotype 5 and genotype 6 in 16 (4.72%), mixed genotype in 25 (7.37%) and untypeable in 32 (0.69%). In control group, 77 (3.7%) individuals was confirmed positive for TTV. Total 0.59% (12) individuals were positive for HBsAg, of which 58.33% (7) were male and 41.7% (5) were female, while HBV DNA was positive in 91.66% (11) individuals. Anti-HCV was positive in 3.2% (65) individuals, of which 58.46% (38) were male and 41.54% (27) were female, while HCV RNA was positive in 69.23% (45) individuals. In high risk group, 973 (4.8%) individuals was confirmed positive for TTV. 3.6% (166) individuals were positive for HBsAg, 64.93% males and 35.06% females, while HBV DNA was positive in 1.6% (75) individuals. Anti-HCV was positive in 17.39% (801) individuals, 18.83% were males and 12.73% were female, HCV RNA was positive in 7.94% (366) individuals. Dual infection was detected in 0.13% (6) individuals. HBsAg were detected in 11.86% (58) thalassemia patients, 11.54% (42) hemophilia patients, 2.75% (22) in patients undergoing hemodialysis and 3.47% (32) of patients with surgery. Anti-HCV were detected in 29.24% (143) thalassemia patients, 43.96% (160) hemophilia patients, 41.93% (335) of patients undergoing hemodialysis and 10.62% (98) of patients with surgery. Co-infection with HBV and HCV was encountered in 0.63% (5) of patients undergoing hemodialysis and 0.11% (1) of patients with surgery. Statistical significance was observed for TTVs between thalassemia patients and hemophilia patients (χ2=24.31, p<0.00001), thalassemia patients and xii surgical patients (χ2= 69.174, p<0.00001), surgical patients and HD, (χ2= 127.594, p<0.00001), surgical patients and hemophilia patients (χ2= 96.835, p<0.0001) and HD and hemophilia patients (χ2= 6.158, p=0.013082). However, the difference between thalassemia patients and HD (χ2= 2.173, p=0.14045) was not statistically significant. Conclusion The present study critically evaluated prevalence of three major transfusion-associated infections, namely infections by HIV, HCV, and HBV. The present investigation showed i. A substantial percentage of the individuals harbored TTV infections (21.21%). ii. The prevalence of TTV was high (34.8%) in high risk group in comparison to the control group (3.7%) and thus ranked at high risk for TTVs infection. TTV Infection prevalence was 9.41 folds higher in high risk group than in control group. It showed that TTVs continue to endanger safe blood supply in a country. iii. The frequency of HCV was high (17.39%) in both high risk group and control group in comparison to HBV positivity (3.6%), the situation is alarmingly and is a matter of concern and effort. iv. The prevalence of untypeable genotypes of HBV and HCV was alarming v. TTV positivity rate increase with increase in the age in high risk group as their transfusion requirements increased with age, thus they were more prone to these infections. vi. The prevalence of mixed genotypes of HCV was matter of concern. vii. This burden of TTV showed that prevention of spread of TTV should be the main goal at the current time. xiii viii. The over flaws in blood transfusion services can be overcome by development of a fair and organized system for safe blood screening, monitoring the implementation strategies for recruitment and retention of safe donors. Recommendation The occurrence of TTV infections should still be monitored carefully to reduce the rate of infections to ensure safer and more reliable blood for transfusion. Education regarding awareness about TTVs, screening strategies, sensitization and vaccination must be carried out to ensure that people are well enlightened and protected from viral diseases. All individuals particularly patients receiving blood transfusion should complete vaccination for Hepatitis B before starting transfusion. Strict and concrete efforts are required to cut down the infection rate through proper screening of blood and blood products, strict emphasis on receiving the vaccine, uniform strict criteria for donor selection.