The defending champions skated a flawless free dance but were edged by American rivals Charlie White and Meryl Davis.
The Canadians finished with 190.99 points after collecting 114.66 in Monday's free dance.
White and Davis finished first with 195.52 points, including 116.63 in the free dance, to become the first American champions ever in the event.
Virtue, from London, Ont., and Moir, from Ilderton, Ont., trailed the Americans by 2.56 points after Sunday's short dance.
Elena Ilinykh and Nikita Katsalapov of Russia took bronze with 183.48.
Kaitlyn Weaver and Andrew Poje of Waterloo, Ont., finished seventh with 169.11 while Alexandra Paul of Midhurst, Ont., and Mitchell Islam of Barrie, Ont., were 18th with 138.70.
Virtue and Moir — two-time world champions —train with Davis and White in Canton, Mich., and share a coach in Russian Marina Zoueva.
Between them, the two North American teams have captured every international ice dance title since Virtue and Moir won Olympic gold four years ago in Vancouver.
The results have swung in the Americans' favour over the last year, however, with Davis and White winning the 2013 world title and the Grand Prix Final in December.
Note to readers: This is a corrected story. An earlier version reported that Weaver and Poje were sixth and Paul and Islam were 17th.