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Aim: A study was aimed to assess the knowledge, value, opinion, and practice regarding the use of dental sealants among private dental practitioners in Chennai, Tamil Nadu, India. Materials and Methods: A self-administrated questionnaire were distributed to 192 private dental practitioners in Chennai, Tamil Nadu, India by using simple random sampling. A convenience sampling technique was employed. The questionnaire consisted of 28 items, which included information about knowledge, value, opinion, and practice regarding dental sealants. The questionnaire was obtained from the study by San Martin et al. 2013 and Kailash Asawa et al. 2014. Frequency distribution was tabulated. For frequency distribution strongly, strongly agree, and agree were combined as “agree” and strongly disagree and disagree were combined as “disagree.” There were no changes in “neutral.” Results: Among the 196 study subjects 56.2% were males and 43.8% were females with their clinical experience of 52.1% for 15 years. The mean scores for knowledge, value, opinion, and practice were 41.8 ± 3.7, 18.7 ± 2.8, 18.1 ± 1.4, and 12.9 ± 2.3, respectively. Conclusion: The results suggest that dental practitioners had satisfactory knowledge about pit and fissure sealant and had neutral attitudes about sealants being effective. Dental practitioners adequately used the pit and fissure sealants but they did not follow the standardized procedures and specific guidelines.