The Mysteries of Laura features Max Santiago, a gay character whose portrayal is consistently positive. His identity is treated with dignity and normalcy, integrated into the narrative without being a source of mockery or negative stereotypes. The show depicts his personal life and professional competence in an affirming manner, contributing to a supportive representation of LGBTQ+ individuals.

The show features a transgender character, Evelyn Reed, in one episode. Her identity is revealed during a murder investigation and is handled factually by the main characters. While her past identity is a plot point, it is not used for mockery or villainy, nor is it the central theme of the episode, resulting in a neutral portrayal.

The American adaptation of the Spanish series "Los misterios de Laura" features characters like Billy Soto and Meredith Bose. These characters, originally portrayed as white in the Spanish source material, are played by Black and Indian-American/Dutch-Indian actors, respectively, in the US version.