Background and Aims: Tuberculosis (TB) and HIV/AIDS have become major public health problems in many countries, including Nigeria. A serious concern is that the TB situation in Nigeria has been worsened by the prevalence of HIV and AIDS. The condition has given rise to a new epidemic of TB and HIV/AIDS coinfection, a combination of two lethal diseases already occurring in epidemic proportions. Aim: This study retrospectively assessed the prevalence of TB and HIV/AIDS co–infection among patients that attended the Directly Observed Treatment Short-course (DOTS) centers in Anambra State, Southeast, Nigeria, between 2013 and 2017. Methods: The study adopted a descriptive and retrospective epidemiological survey design. A total of 1443 case files of patients aged 15 to 60 who were treated in DOTS centers selected from Anambra State's 21 Local Government Areas between 2013 and 2017 were investigated. The uniform data form, a standardized instrument used in Anambra State's health facilities for data collection, was used to collect data from case files of all those identified as co–infected with TB and HIV/AIDS. Results: The mean prevalence rate of TB and HIV/AIDS co–infection in the state during the five-year period (2013–2017) was 20%. The highest annual prevalence of TB and HIV/AIDS co–infection was recorded in 2014 (23.84%). The state's prevalence of TB and HIV/AIDS co–infection increased dramatically from 13.17% in 2013 to 23.84% in 2014, followed by a slight downward trend to 22.80% in 2015, 20.17% in 2016, and 20.03% in 2017. The annual prevalence rate of TB and HIV/AIDS co–infection was significantly different (p<0.05) over the five-year period. However, there was no statistical difference in the annual prevalence rate between 2014 and 2017 (p>0.05). In terms of gender, age, marital status, and occupation, females (59.5%), those aged 15 to 25 years (30.7%), married people (43.9%), and traders/business owners (50.7%), respectively, had the highest rates of tuberculosis and HIV/AIDS co–infection during the study period. Conclusion: The findings of this study show that young people, females, married people, and traders/business owners appear to be the most vulnerable groups affected by TB and HIV/AIDS co–infection. This accounts for the vast majority of the disease burden in Anambra State, Southeast, Nigeria. To address the high prevalence of TB and HIV/AIDS co–infection in Anambra State, novel intervention and control programs should be developed and implemented. Existing interventions should be strengthened to achieve more robust and effective preventive measures to address the state's high prevalence of TB and HIV/AIDS co–infection rates.