Despite extensive use of organochlorinated pesticides (OCPs) such as dichlorodiphenyltrichloroethane (DDT) in Italy in the 1940s to 1970s, especially for public health control of malaria mosquitoes, information on their exposure levels among the general population is limited. These OCPs can be a source of health risk to human. A total of 137 blood samples were collected from residents of the general population of three Italian towns, Novafeltria, Pavia and Milan, to determine the levels of eight OCPs in blood serum. The concentrations of beta-hexachlorocyclohexane, hexachlorobenzene (HCB), 1,1-dichloro-2-( o-chlorophenyl)-2-( p-chlorophenyl)ethylene, 1,1-dichloro-2,2-bis( p-chlorophenyl)-ethylene ( p, p' -DDE), 1,1-dichloro-2-( o-chlorophenyl)-2-( p-chlorophenyl)ethane, 1,1-dichloro-2,2-bis (4-chlorophenyl)ethane, 1,1,1-trichloro-2-( o-chlorophenyl)-2-( p-chlorophenyl)-ethane and 1,1,1-trichloro-2,2-bis( p-chlorophenyl) ethane were measured by gas chromatography–mass spectrometry. Variations in serum concentrations of OCPs with respect to place of residence, gender, age and body mass index (BMI) were evaluated by non-parametric tests. p, p' -DDE and HCB were the most abundant and major contributors of total OCP concentration. Their levels differed significantly between the three towns with a trend Milan > Novafeltria > Pavia ( p < 0.0001). Females had significantly higher concentrations of HCB and p, p'-DDE than males in the overall population sample. HCB concentrations were significantly higher in females than in males of Milan ( p = 0.029). We observed positive correlations of p, p'-DDE and HCB with age in Novafeltria subjects ( r = 0.468, p = 0.004). Total OCP concentrations differed significantly across BMI categories ( p = 0.018) in overall population. We have demonstrated a clear pattern of the main OCPs in a fairly large population. Generally, our study provides information on OCPs exposure among the Italian general population and provides indications for further investigations.