The necessity and importance of blood donation well known like in thalassemia, leukemia, lymphoma, patients on chemotherapy and after massive hemorrhage from trauma etc.but it’s transfusion can also lead to some major health problems like transmission of infectious diseases such as syphilis and so on. To prevent transmission of infectious diseases through blood transfusion its screening is performed everywhere. For this purpose, a research study is carried out during January 2019 to April 2019 at Mardan medical complex and Mardan teaching institute (MMC-MTI) Mardan KP, Pakistan. The specimens were collected from 625 blood donors with 619 males and 06 females. The samples were sent to the serology section of the pathology department for the diagnosis of syphilis through architect syphilis TP. The sero-prevalence of syphilis and the variation of prevalence in the previous and recent study were the objective of this research. It has found that only 06 male individuals are found to have syphilis infection while there is no such case reported in female individuals. So the overall sero-prevalence and sero-prevalence in male individuals were the same as 0.96%. The most positive cases were in the age group 38-74 years of age individuals. Which is the most sexually active age and it is reported that 1.8% of the sero-prevalence is found. The seroprevalance of syphilis in A- blood group was 5.5%, B+ patients had 1.56%, and with O+ patients it was 1.16%. From the results and analysis, the sero-prevalence of syphilis in the area and population where the study performed are at more risk of syphilis because per annual 0.30 increases is recorded by comparing the recent results with previous studies. As with the passage of time people are educated so we cannot say that it is because of the lack of awareness no also. So the opinion is that the lifestyle, social interaction of people with one another and carelessness in sexual intercourses is the main risk factors responsible for this increase in sero-prevalence of syphilis.