Brucella infections (Brucellosis) is a disease that reaches individuals who work directly in contact with animals. The aims of this study was to investigate the prevalence of Brucella species in dairy women seller and their families by Real Time quantitative- qPCR reaction. The study was done in AL-Kut city (Iraq), Serum samples were collected from some women who sell dairy products (hawkers) in city markets and resident in different four villages around city, As in addition their families who live contact with cattle and participated in make the dairy products. One hundred eighty six (30 dairy women seller +156 of their families) Serum samples were collected between July 2015 to November 2015, and the Rose Bengal test (RBT) was performed on each sample.A total of 30 dairy women seller (DWS) were 6 (20%) Brucella seropositive, out of 156 persons (families of dairy women seller) were 44 (28%) Brucella seropositive. All seropositive test by Real Time- PCR reaction, PCR was based on unique genetic loci of B. melitensis and B. abortus. The regions were chosen for the construction of primers and probes for species differentiation: BMEII0466 gene for B. melitensis and Bru-Ab2_0168 gene for B. abortus. Out of (50) Seropositive B. meltitensis was diagnosed in 27(54%) sample , B. abortus in 11(22%). sample . Among the (20) women (6 DWS+14 of their families) were seropositive. The questionnaire showed 10 (50%) of them had aborted and 2 (10%) presented with a stillbirth. Human brucellosis appears to be of public health importance in Iraq and more attention should be drawn on the disease. The current study provides a basis for larger studies to establish the incidence of Brucella infections among rural population in Iraq.