Background: Mycobacterium Tuberculosis (TB) has been an important cause of mortality and morbidity all over the world but is specifically affecting developing countries like India where the disease is endemic. Inspite of being asymptomatic, female reproductive system is found to be very vulnerable to tubercular infections and by the time, Nontuberculous mycobacterial infections are also emerging and are found to cause serious genital infections in the females of child bearing age leading to infertility. Objectives: Detection of Mycobacterial (MTB & NTM) infections among infertile females via different techniques like MRT-PCR, staining and their isolation by liquid media (using BACTEC 320). Material and Methods: A total of 217 samples were processed involving the isolation of both (MTB and NTM). All three techniques were processed staining, culture and MRT-PCR to find the prevalence and efficacy of the techniques. For NTM and MTB differentiation SD MPT 64 card test was done. Results: Out of 217, TB suspected infertile female’s maximum lies under the age group of 26-30 i.e. 34.10% (approximately 74 females) and the lowest were under the age group of 15-20 years i.e. 2.3% (5 females). There were total 24 NTM MRT-PCR positives and 29 MTB positives. So the total number of female patients who were positive by MRT-PCR technique was 53, whereas in liquid culture there were 23 positives (12 NTM & 11 MTB). 4 samples were found to be AFB smear positive by ZN staining. Conclusion: As Mycobacterium tuberculosis remains one of the leading causes of female infertility. According to a few studies, NTM’s were found to be increasingly important pathogens causing male genital infections and infertility, but this study gave an insight that NTM’s can even lead to female infertility and infections.