Detection of Anaplasma organism in Bangladesh is mostly performed by microscopic examination which would make poor detection of organism and less specific identification as well. The aim of this study was to detect Anaplasma marginale from both tick and cattle reared in Dinajpur and Sirajganj district of Bangladesh by PCR. From February 2017 to February 2018, blood and tick samples were collected from 370 cattle. The blood samples were initially screened for Anaplasma spp by staining of blood smear with Giemsa stain and examined under microscope. The DNA was extracted from the microscopically positive blood samples and subjected to Polymerase Chain Reaction (PCR). In the first step, PCR was performed targeting the 16srRNA gene sequence to confirm the presence of Anaplasma organism in the samples (PCR product size 781bp). In second step, the presence of A. marginale organism was detected in the samples targeting the MSP1β gene of A. marginale (PCR product size 95bp). The overall prevalence of tick infestation in cattle was 59.20% (219/370 cattle). A. marginale was detected by nPCR in 33.78% blood samples of cattle and 17.35% tick samples. Among the ticks 25 Rhipicephalus (Boophilus) microplus and 13 Haemaphysalis bispinosa, were positive with A. marginale. The study indicates that A. marginale is common in cattle of Dinajpur and Sirajganj district of Bangladesh with higher infection rate in Sirajganj district.