Background: Tuberculosis is caused by Mycobacterium tuberculosis and is spread mainly through contact with air droplets and respiratory fluid from an infected person. Drug of choice for its treatment are rifampicin and isoniazid respectively. However, in recent times, resistance to these drugs as with other antibiotics has been observed across the globe. This study was thus aimed at determining the prevalence of rifampicin resistance strain among TB patients attending two major hospitals (Braithwaite Memorial Hospital and University of Port Harcourt Teaching Hospital) in Port Harcourt. Method: Sputum of patients was collected, tested for the presence of TB and rifampicin resistance of the isolates were determined using GeneXpert assay approach. Result: A total of 158 isolates from patients who were TB positive were tested for rifampicin resistance. 13.3% of these were resistant to rifampicin drug. 8.9% was from UPTH while 4.4% was from BMSH. 9.5% were less than age 45 while 3.8% were greater than or equal to age 45 (≥45). Males made up 8.2% of the total prevalence while females made up 5.1% of the total prevalence. Alternative drugs to rifampicin and isoniazid drug should however, be prescribed in confirmed cases of resistance outcome in our health facilities especially in the rural communities. Conclusion: Nonetheless, the use of antibiotics indiscriminately without the effective laboratory assay and physician’s prescription should be discouraged at all levels and patients receiving treatment should be monitored to adhere strictly to the desired dosage as non-adhering to the gold standard option may lead to drug resistance outcome among vulnerable patients who are literally exposed to lack of health education and functional health facilities in the remote communities which are also hard to reach area due to the environmental terrine.