The chili pepper (Capsicum spp. L.) is a vegetable of economic importance that has been affected worldwide by the cucumber mosaic virus (CMV), a pathogen that causes a devastating disease in this crop. The aim of this research was the detection and characterization of CMV in chili pepper crops in Valle del Cauca, Colombia. Leaves of three chili pepper varieties (tabasco, cayenne and habanero) with viral symptoms were collected in four municipalities of Valle del Cauca. Total RNA was purified and a fragment of capsid protein (CP) from CMV was amplified by RT‑PCR. Then, it was sequenced and bioinformatically analyzed, and from these sequences, specific primers were designed. From 71 chili pepper samples collected in Palmira, Yumbo, Vijes and Yotoco, 37 were positive for CMV (52.1%). The CMV chili pepper sequence analysis showed that they had their highest identity (98.5%) with a CMV isolated from bananas in Ecuador. Specific primers designed for CMV chili pepper showed greater sensitivity for detecting this virus (64.7% vs. 52.1%). The CMV chili pepper CP analysis indicated that it could be transmitted by the species Aphis gossypii. This r the first time, the molecular characterization of CMV in three chili pepper varieties.