Aims: The study aims to assess the level of awareness regarding Hand hygiene among the population in urban and rural areas of South India.
 Study Design: A web based cross sectional survey.
 Place and Duration of Study: An online study was conducted among public of South India for a period of 6 months.
 Methodology: The data was collected through online mode by providing google link to fill questionnaire form through various social media platforms. The questionnaire consists of demographic details, knowledge and practice related questions. SPSS was used to interpret data and chi-square test, independent T test, backward multiple regression analysis was done.
 Results: From the collected data a total of 1178 responses were collected (urban:726, rural:452). From independent T test the knowledge mean score of HH was: 10.07±2.136 for urban population and 9.79±2.138 for rural population. By this urban population has more knowledge than rural population. From backward multiple regression analysis of Knowledge score, the findings shown that the urban male residents with 2 members household residing in Tamilnadu have preferably more knowledge than others when compared with demographic details. From the independent T test the HH practice mean score was 11.25±1.991 for urban residents and 10.77±2.280 for rural residents. From the Backward multiple regression analysis of Practice score, the findings shown that most parsimonious combination of region (rural, urban), age group (10-30 years, 30-60 years and >60 years), gender(female, male), state and UT (Andhra Pradesh, Karnataka, Tamilnadu, Kerala, Telangana, Andaman and nicobar, Puducherry, Lakshadweep), household (2, 3, 4, >4 members), education level (10 and below 10, intermediate, UG, PG, Ph. D) in predicting the practice score.
 Conclusion: The study results concluded that urban residents have preferably more knowledge than rural residents. Creative campaigns and awareness programs should be conducted to attain persistent improvement in HH practices.