Background and Objective: This study aimed at determining the prevalence of Mycobacterium tuberculosis and non-tubercular bacteria co-infection among tuberculosis patients attending Mile four hospital, Abakaliki, Ebonyi State, Nigeria. The study was carried out in Microbiology laboratory of Alex Ekwueme Federal University Ndufu-Alike Ikwo, Ebonyi State. Methods: The samples were analyzed macroscopically for the presence of pus and blood and were cultured on chocolate agar using the streaking technique. Cultures were incubated for 24hours at 37. Bacterial growth after incubation were identified based on colonial characteristics, Acid Fast staining and Gram staining techniques and other biochemical tests. Results: The study revealed that the prevalence of co-infection between M tuberculosis and other bacteria pathogens is 4%. Out of the eight bacterial isolated from the different samples evaluated, Escherichia coli had the highest frequency and percentage occurrence of 5(25%), followed by Staphylococcus pyogens 3(15%), Haemophilus influenzae 2(15%), proteus 1(15%), Streptococcus pneumoniae 3(10%), Pseudomonas spp 2(10%), while Klebsiella spp has the lowest number of occurrence with frequency and percentage of 1(5%) and Staphylococcus aureus also had the lowest number of occurrence with frequency and percentage of 1(5%). It was observed that Mycobacterium tuberculosis and Escherichia coli co-infection had the highest occurrence from the study. This present study has revealed the prevalence of co-infection among M. tuberculosis and other bacteria pathogens among TB patients. The result of the sensitivity shows that oflaxacin, pelfloxacin and cephalosporin have an inhibition zone diameter of 18mm respectively why penicillin, pelfloxacin, septrin have the highest zone of inhibition diameter of 20mm. Conclusion: However, findings from this study will assist the concern health authority on the needs to intensity preventive and control measure against the treatment and management of tuberculosis.