Anaplasmosis is a widespread vector-borne disease affecting dogs, and Anaplasma platys is the major etiological agent of the disease. The study examines anaplasmosis molecular prevalence, related risk factors, and alteration of hematological variables in Anaplasma-affected dogs. A total of 150 blood samples were collected from dogs in the district of Lahore, Pakistan. The samples were screened with PCR targeting the 16S rRNA gene of Anaplasma. Sequencing of samples that were found positive after performing PCR was conducted. A questionnaire was developed to collect epidemiological data on subject dogs, and the information was analyzed with a logistic regression model using SPSS. The current study revealed an 11.34% (17/150) prevalence of anaplasmosis in dogs based on PCR detection. Tick infestation, previous tick history, house hygiene, and tick control status were major risk factors linked with disease occurrence. Red blood cell count, packed cell volume, hemoglobin, and platelet count were decreased significantly (P < 0.05) in Anaplasma-infected dogs. Phylogenetically, the 2 isolates of the current study clustered together, and that cluster was very similar to A. platys isolates from India, Malaysia, and Thailand.