Ardoin was born near Basile in Evangeline Parish, Louisiana a descendant of both free and captive people. Amadee spoke only French and did not speak English, as did most people in this French speaking region. Developing his musical talents in preference to undertaking farm work, he played at dances, often for Cajun audiences, with fiddle players Alphonse LaFleur and Douglas Bellard. He moved around the area frequently, settling at one point near Chataignier where he met Cajun fiddle playerDennis McGee. They established a more regular musical partnership, playing at local house parties, sometimes attended by Ardoin's young cousin, Alphonse "Bois Sec" Ardoin. Ardoin and McGee were among the first artists to record the music of the Acadiana region of Louisiana. On December 9, 1929, they recorded six songs for Columbia Records in New Orleans. They made further recordings together in New Orleans in 1930, and in San Antonio, Texas in August 1934. Ardoin also made solo recordings in New York City in December 1934. The recordings were issued on various labels, including Brunswick, Vocalion, Decca, Melotone and Bluebird. In all, thirty-four recordings with Ardoin playing accordion are known to exist. His recordings and performances became popular throughout southern Louisiana. In the late 1930s, he played regularly in Eunice, Louisiana with fiddle player Sady Courville, but the two did not record together. Ardoin's music combined "European song forms and African rhythmic approaches such as swing and syncopation... personified this cultural blend and enhanced its development through his deft technique and his ability to improvise. Ardoin was a lively, inventive accordionist who could keep a crowd dancing while playing alone. He was also a soulful singer whose emotional style made dramatic use of elongated, high-pitched notes."
The circumstances that led to Ardoin's death, and the final cause of his death, are uncertain. Descendants of family members and musicians who knew Ardoin tell a story, now well-known, about a racially motivated attack on him in which he was severely beaten, in about 1939, while walking home after playing at a house dance near Eunice. The most common story says that some white men were angered when a white woman, daughter of the house, lent her handkerchief to Ardoin to wipe the sweat from his face. According to Canray Fontenot and Wade Fruge, in PBS's American Patchwork, Ardoin left the place and was run over by a Model A car which crushed his head and throat, damaging his vocal cords. He was found the next day, lying in a ditch. According to Fontenot, he "went plumb crazy" and "didn't know if he was hungry or not. Others had to feed him. He got weaker and weaker until he died." Others consider this story apocryphal. Other versions say that Ardoin was poisoned, not beaten, possibly by a jealous fellow musician. Contemporaries said that Ardoin suffered from impaired mental and musical capacities later in his life. Some recent studies have concluded that he died as a result of a venereal disease. He ended up in an asylum in Pineville, Louisiana, where he was admitted in September 1942. He died at the hospital two months later, and was buried in the hospital's common grave. On March 11, 2018, a life-sized statue of Ardoin was unveiled at the St. Landry Parish Visitor Center. The project was headed by Louisiana's former Poet Laureate, author, and professor Darrell Bourque, who wrote a book of poetry titled 'If You Abandon Me: An Amédé Ardoin Songbook', the cover of which features artwork by Pierre Bourque.
