This study aimed to determine the Mycobacterium tuberculosis (MTB) in HIV seropositive patients with cervical lymphadenopathy using different diagnostic tools. A hospital- and laboratory-based, cross-sectional descriptive study was conducted in 2014-2015. A total of 60 cervical lymph node aspirates from the Thaketa Specialist Hospital were tested for MTB using different diagnostic tools which include the conventional methods; Ziehl-Neelsen staining for acid-fast bacilli (AFB) smear and culture on Lowenstein-Jensen medium and the molecular methods; polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP). On macroscopic examination of lymph node aspirate, 55 cases (92%) had caseation and 5 cases (8%) had clear fluid. In microscopic examination, 29(48.3%) cases were positive for AFB and 31(51.7%) were negative. In culture, MTB were identified in 31(51.7%) cases but 29(48.3%) were negative. In PCR, 45(75%) cases were detected for MTB although 15(25%) were negative. In LAMP, 45(75%) cases were positive for MTB but 15(25%) were negative. Regarding culture as a gold standard, sensitivity and specificity of AFB-smear, PCR and LAMP were 84% (95%CI=66-95%) and 90% (95%CI=73-98%), 90% (95%CI=74-98%) and 41% (95%CI=2461%), and 81% (95%CI=63-93%) and 31% (95%CI=15-51%), respectively. Although molecular methods i.e. PCR and LAMP are useful with high sensitivity (90% vs. 81%), respectively and they can identify scanty amount of TB-DNA, AFB-smear (sensitivity 84%) with macroscopic visible caseation is the most reliable diagnostic method which is simple, cheap and generally available at district hospitals in resource limited settings.