Avian influenza virus (AIV) subtype H9 carries great zoonotic potential and human infections have been reported in influenza endemic countries. To the best of our knowledge, very little is known of seroprevalence of H9 in human population in Pakistan. The main intention with which the current study conducted to report the prevalence of the H9 virus among the poultry workers and to add a new piece of knowledge to its prevalence in Pakistan which is the first step taken for before control measures can be executed.Overall 325 sera samples were obtained from poultry encounterd population and a general publicfrom January 2018 to September 2018 in Rawalpindi, Pakistan. Haemagglutinin-inhibiting (HI) assay was performed to determine the seropositivity for serum samples. Overall seroprevalence level of anti-H9 antibody titres was 12 % (39/325) in occupational individuals and general population (control group). Anti-H9 antibody titres was 8.8%, 8.% and 34.1% in age groups of 16-39, 40-59 and >60 years respectively of poultry-exposed workers. On the other hand, anti-H9 antibody titres of general population was 25% and 16.6% in age groups of 16-39 and 40-59 respectively. Interestingly, anti-H9 antibody titres in the general population among 16?39 was 25% and among 40?59 was 16.6%. The highest percentage of sero-positivity (13.7%) of antibody titer to H9N2 was found in the poultry butchers followed by poultry farm workers 11.4%. The lowest seropositivity (7.1%) was recorded in veterinarian. Serum samples from males revealed significantly higher anti-H9 antibody titre (12.4%) than females (8.5%).It revealed that antibody titer of butcher, had significant (P<0.05) difference from antibody titer of poultry salesmen and veterinarians but non-significant difference from poultry farm workers. Antibody titer of butcher had differed significantly (P<0.05) as compared to group of general population. In the control group (general population group), 17 individual (68%) had no detectable antibody titer against H9N2 and only 12% people had antibody titer of = 1:40. To recapitulate, our findings revealed poential transmission AIV H9 subtype from avian to humans and highlighted that poultry professional linked people are at great risk of AIV infections than general population. Hence, further detailed and more systemic approaches to study seroprevalence ofH9N2 AIV infections in human and live poultry population in all over Pakistan is recommended to determine the relations between clinical significance and seropisitivity in humnas and identify possible solutions to reduce the transmission of AIV in poultry workerfrom live poultry.