Ehrlichia are gram-negative, pleomorphic, and obligate intracellular bacteria that infect many varieties of mammals; particularly Ehrlichia canis has been identified as a causal agent of canine monocytic ehrlichiosis (CME). In Colombia, the disease diagnosis is based on treatment response, immunochromatographic techniques, and microscopic form findings compatible with hemoparasites; these tools do not guarantee a precise diagnosis even when there is no certain clarity about agent typification. The aim of this research was to evaluate, using PCR test, the potential presence of E. canis in blood samples of dogs that reside in Ibague (Tolima) with signs compatible with CME. For this purpose, PCR touchdown was standardized to identify 16SrRNA gene; this procedure was applied in 50 canine blood samples (40 animals with clinical signs compatible with E. canis and 10 without it). This research is a non-experimental cross design. Hematological findings revealed that the 6% (4/50) had normal results, the remaining animals had some change in blood cells; furthermore, the study revealed that 7.5% of dogs with clinical signs (3/40) had normal blood values, and 10% of control animal group did not have changes in blood parameters (1/10). There were no findings of E. canis morula in blood films. Afterward, the molecular analysis established that 60% of dogs were PCR E. canis positive. This research is one of the first E. canis molecular detections in Colombia, which can be the basis for future research that strengthens the diagnostics and characterization of diseases with epidemiological impact in human and animal health.