Even though Romney still holds strong advantages in financing and organization, his campaign will now have to refocus to fight back the challenge from the surging conservative Santorum.
Backed by a wealthy "Super PAC" that pays for attack ads against rivals, Romney had excelled in major contests thus far in the race. After big wins in Nevada and Florida in the previous week, he did little campaigning in Minnesota and Missouri and had been expected to win easily in Colorado.
Until Tuesday, Santorum had won only one of the first five Republican contests in the state-by-state battle for the Republican nomination to face President Barack Obama in the Nov. 6 election.
But on the first day of multiple nominating contests in the 2012 primary season, Santorum trounced Romney by 30 percentage points in Missouri. That vote was a non-binding primary, but has symbolic value as a measure of support in a big Midwestern state.
In Minnesota's caucuses, Santorum won with 45 percent of the vote. But in another setback to former Massachusetts governor Romney, US congressman Ron Paul was in second place with 27 percent and Romney was a distant third with 17 percent.
The race was closer in Colorado, but Romney had been expected to win easily. But Santorum won by 5 percentage points over Romney.
With eight contests to date in the Republican race - including the three on Tuesday - Santorum has now won in four states, Romney in three and former House of Representatives speaker Newt Gingrich in one.
The former Pennsylvania senator is a devout Catholic who had been in a battle with Gingrich to become the conservative alternative to the more moderate Romney. Gingrich was not on the ballot in Missouri and was crushed in the other two states.
Santorum's victories give heart to social conservatives fighting battles of abortion, gay marriage and contraception in recent days.
The Minnesota result marked the first time so far in the 2012 Republican race that Romney did not come in first or second. Romney also lost in two states - Colorado and Minnesota - that he won in his failed 2008 bid for the Republican presidential nomination.
"Wow. Conservatism is alive and well in Missouri and Minnesota," Santorum told supporters in St. Charles, Missouri after results were tallied in those states but before the Colorado numbers came in.
"I don't stand here to claim to be the conservative alternative to Mitt Romney, I stand here to be the conservative alternative to Barack Obama," he added to cheers from the crowd.
The startling results in Colorado, Minnesota and Missouri raised fresh doubts about whether Romney, long considered the most likely Republican nominee, can win over the broad swathe of Republican voters, particularly the most conservative ones who are the most unenthusiastic toward him.
"This was a good night for Rick Santorum. I want to congratulate Senator Santorum and wish him the very best. We'll keep on campaigning down the road, but I expect to become our nominee with your help," Romney told supporters in Denver.
Santorum has rebuilt a political career left in tatters in 2006 when voters in Pennsylvania threw him out of the Senate by an 18-percentage point margin. But he still faces an uphill battle for the presidential nomination, considering Romney's vast organizational and financial advantages.
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