Aim: To evaluate the role of implementing sterility techniques for prevention and transmission of infections in Labor Room. Methods: The prospective observational study was conducted in the reputed hospital of Kota Rajasthan, India. This study was done at labor room through implementation of infection control rules. To assess the efficacy of infection control practices various factors were considered and analyzed before and after implementation of infection control policies and practices. Various aspects observed were environmental surveillance reports, clinically suspected or confirmed infections of patients in labor, needle stick injuries incidence, puerperal infections rate, neonatal infections, hand hygiene compliance. Results: Environmental surveillance was done by sending swabs from labor room to microbiology for sterility check by culture. 8 were reported as positive for bacteria before implementing policies, whereas no organisms were reported after. Needle stick injury exposure also decreased from 15% to 4% members among staff involved in labor room works. Number of Clinically suspected or confirmed infection patients were almost same around 60% before and 40% after implementing policies, but the incidence of puerperal infections and neonatal infections reduced by proper following of implemented policies. Puerperal and Neonatal infections were noted as 33% and 39% before, 11% and 16% after introducing proper infection control guidelines. Hand hygiene compliance was observed randomly among different staff members including doctors, interns, nurses, students, biomedical management staff. Compliance decreased from 25% to 5% after introducing mandatory hand hygiene guidelines. Adherence to guidelines of personal protective equipment were observed in labor room. 40% and 81% were using PPE according to protocol before and after implementing protocols respectively. Conclusion: We concluded that the health care personnel are required to adhere to standard precautions and infection control guidelines and all should undergo pre employment check up to avoid transmission of infections. Participation in at least one Infection control education activities has to make compulsory for all health care personnel.