INTRODUCTION: Diagnosis of tubercular lymphadenitis is daunting as there are varied clinical presentations and no single confirmatory gold standard test. Cartridge-based nucleic acid amplification test (CBNAAT) of the lymph node is a rapid molecular diagnostic test for simultaneously detecting tuberculosis (TB) and rifampicin resistance. AIM: To evaluate the performance of the CBNAAT test for detecting M. tuberculosis in lymph node specimens compared to fine needle aspiration cytology (FNAC). MATERIALS AND METHODS: The study was conducted in a rural tertiary care hospital in central India. A total of 180 patients clinically suspected of tubercular lymphadenitis were included. The male-to-female ratio was 1:1.3. The average age was 33.3 years. The age group 21–40 years had the highest number of cases. The most common complaints among the patients were fever (29.4%), followed by loss of appetite (9.5%), weight loss (9.5%), and cough (6.6%). However, most patients presented to the hospital with only lymphadenopathy (44.4%). The most common site involved was the anterior cervical lymph node (78.8%), followed by the axillary group (10.5%), submandibular (2.8%), inguinal (2.8%), supraclavicular (2.2%), submental (1.7%) and infraclavicular (1.1%) group of lymph nodes. The patients were subjected to both FNAC and CBNAAT testing. Results were reported as positive or negative for M. tuberculosis as CBNAAT gives a semiquantitative estimate of the concentration of bacilli. Rifampicin resistance results were reported as detected or not detected. RESULTS: Cytological examination of the lymph node aspirates revealed that most were tubercular lymphadenitis cases. Cytomorphological analysis of the cases of tubercular lymphadenitis revealed Type 6 (tubercular abscess) as the predominant pattern. CBNAAT testing detected 26 cases of M. tuberculosis and three cases of rifampicin resistance. The study reported a specificity of 92.92% and low sensitivity of 26.86% of combined FNAC and CBNAAT is much higher compared to only CBNAAT. CONCLUSION: CBNAAT, along with FNAC, is a valuable addition in first-line investigations of tubercular lymphadenitis to make a timely diagnosis.