INTRODUCTION Immunization is one of the cost effective intervention for prevention of major illness in child. Only availability of vaccines is not sufficient, there are various other factors which influences the effectiveness of immunization. These factors are proper storage, transportation, administration along with knowledge and practices of health care worker. MATERIALS AND METHODS The present cross sectional study was conducted in the rural area of Chirang district, Assam in January 2015 to May 2015. 23 ANM at the session site were interviewed with the help of pre tested structured schedule and observation method. RESULTS Knowledge regarding dose and route of vaccines is 100%, knowledge regarding age at which 2 nd dose of measles is given 86.95%, knowledge about freeze sensitive vaccine (DPT, TT) is 86.95% whereas Hep. B is 100%. Practice of hand washing was 65.21% and practice of delivering 4 key messages were 73.91%. Practice of segregation of waste in red and black bag was 65.21%. CONCLUSION Knowledge gap were observed specifically for cold chain, open vial policy, heat and freeze sensitive vaccines. Flaws in practice of immunization were observed as for delivering 4 key messages and hand washing. Regular supervision and training can fill up the gap both in knowledge and practice.