Canine herpesvirus-1 (CHV-1) is recognized to be enzootic in the dog population with a widespread distribution. This pathogen leads to a lethal generalized illness in newborn puppies and is associated with reproductive disorders. CHV-1 should be considered as an important pathogen of neonatal death and infertility; so, it appears to pose a threat for breeding kennels. Although serologic data point to the circulation of CHV-1 among dogs of Iran are available, no definitive diagnosis has been conducted based on the molecular assay. So, this research was done to detect the prevalence of CHV-1 in dogs of Kerman. In this study, the presence of CHV-1 in vaginal specimens and biopsies of the uterus of dogs referring to the Veterinary Hospital of Shahid Bahonar University of Kerman was determined. Fifteen uterine samples and seven vaginal samples were included in group of the pregnant dogs. Moreover, thirteen uterine samples and twenty vaginal samples were related to the dogs displaying suspicious clinical signs such as reproductive disorders. Samples were collected and evaluated using real-time PCR. Viral DNA was detected in 21 samples from a total of 140 (15 %) collected samples which were related to 14 uterine samples (20 %) and 7 (10 %) vaginal specimens. The association of this virus with age, breed, housing, pregnancy and reproductive disorders was not significant. Five positive reproductive samples were belonged to the dogs with a history of reproductive disorders including pyometra, metritis, stillbirths, vaginitis and vaginal prolapse. This study is the first molecular detection of CHV-1 in reproductive samples of dogs in Iran. Considering the significant prevalence of this virus, it is necessary to carry out management measures in controlling and preventing this disease. Tracing CHV-1 requires further research on this virus in dogs of this region.Copyright © 2020 Elsevier Ltd. All rights reserved.