Background: The goal of the study was to investigate the burden of transfusion-transmitted infections (TTIs) hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), syphilis, and malarial parasite (MP) in ABO Blood Groups and Rh Type System among voluntarily blood donors in Khyber Pakhtunkhwa (KPK), Pakistan. It is a retrospective single center cross sectional study. This study was conducted from June 2020 to September 2021 (16 months) at the frontier foundation thalassemia center Peshawar KPK. Donors were physically healthy and fit for donation. Donors with physical disabilities and/or having co-morbid conditions were excluded from the report. Methods: All the samples were screened for anti-HIV, anti-HCV, HBsAg, Syphilis, and Malarial Parasite via ELISA kit and Immune Chromatographic Technique (ICT), respectively. A total of 6311 blood donations were evaluated. The majority of the donations (92%) were from (VNRBD) voluntary non-remunerated blood donation, while only 8% came from replacement donors. Results: Amongst 6311 blood donations, 1.50 % (n=95) were infected at least with one pathogen, HBV positive cases were 0.855 % (n=54), HCV positive cases were 0.316% (n=20), syphilis positive were 0.30% (n=19) and MP positive cases were only 0.031% (n=2). HBV, HCV, syphilis and malaria infections rates were found to be low as compared to the previous data published, while no case was reported for HIV. The study also revealed the distribution pattern of the aforementioned pathogens in blood groups and the Rh type system of the reactive samples. Conclusion: The lower reported in our study indicates the awareness among the people of Peshawar about TTIs and their precautions. The prevalence rate that we are reporting is less than previously published articles in the same domain. result: Amongst these donations, 95 were infected at least with one pathogen, while 6216 were cleared for transfusion; among which HBV positive cases were 0.855 % (n=54), HCV positive cases were 0.316% (n=20), syphilis positive were 0.30% (n=19) and MP positive cases were only 0.031% (n=2). HBV, HCV, and syphilis infections rates were increased while little changes were observed in the malaria infections rates, and no case was reported for HIV. other: No