Introduction: Co-infections during Tuberculosis infection are among the leading causes of morbidity and mortality worldwide. These infections are a major public health problem among patients in most developing countries, including Cameroon. One of the important public problems in coendemic areas is co-infection with TB in humans. Moreover, there is no adequate research hitherto that provides information on the level of co-infection in Tuberculosis patients. Therefore, this study aimed to assess the prevalence and intensity of co-infections in newly diagnosed Tuberculosis patients prior to their enrollment to the Direct Observed Treatment (DOT) program in Buea, Limbe and Douala Tuberculosis treatment centers. Methods: This cross-sectional study was conducted from March to November 2020 using a purposive sampling technique. A fresh sputum sample was collected, processed, and examined through a smear microscopy and culturing technique. Blood samples were collected through finger pricking for the detection of blood parasites using thin and thick blood smears. Stool samples were also collected and processed using a direct wet mount, and Kato-Katz and Modified Ziehl Neelsen staining methods. The data was analyzed using SPSS version 20.0. A Chi-square test was done to assess the prevalence of co-infection among all TB patients recruited for this study. Results: A total of 200 newly diagnosed TB-positive patients were recruited from the three study sites. Out of the total participants recruited, 29.0% were diagnosed positive for blood parasites, 2.0% for intestinal parasites and 15.0% for viral infections, bringing the overall prevalence of co-infection to 46.0%. When specific infections were considered, malaria, filaria, intestinal parasites, HIV and HBV constituted 28.0%, 1.0%, 2.0%, 14.5% and 0.5% respectively. When dual-infection was considered, the results were 0.5% malaria/filaria, 1.5% malaria/intestinal parasites and 2.0% malaria/HIV. The mean parasite densities for all samples diagnosed positive for malaria, filarial and intestinal parasites were 107.0 ± 338.8 parasites/uL, 25.7 ± 356.4 mf/mL and 85.0 ± 85.4 EPG, respectively. Conclusion: The results indicate that there is an overlap of co-infections in TB patients with their interactions expected to lead to complications and difficulty when managing the TB patients, calling for more research and recommending more assessment for co-infection in TB patients before starting treatment.