The team of Calgary's Allison Beveridge, Vancouver's Laura Brown, Jasmin Glaesser, of Coquitlam, B.C., and Kirsti Lay, of Medicine Hat, Alta., finished the race in a Games record time of four minutes 19.664 seconds in the final.
The U.S. earned the silver in 4:26.426 while Mexico beat out Colombia to capture the bronze medal.
The Canadian women have reached the podium in every World Cup or world championship since winning bronze at the 2012 London Olympics.
Calgary's Monique Sullivan added another gold for Canada on Friday, finishing first in the women's keirin final.
Sullivan's gold was the fourth medal for the Canadian squad at the Cisco Milton Pan Am Velodrome. The men's and women's sprint teams both won gold Thursday night.
Sullivan and Kate O'Brien, also of Calgary, won the women's sprint gold medal.
Remi Pelletier-Roy of Longueuil, Que., finished fifth in the men's omnium following the points race, a gruelling 160-lap, 40-kilometre competition.