COLUMBUS, OHIO — Since the turn of the millennium, “Dos a Cero” had become the refrain for quadrennial World Cup qualifying clashes in central Ohio between the U.S. and Mexican national soccer teams.

But after four consecutive home victories by the same two-goal margin against their bitter rivals, the Americans crashed to defeat Friday, 2-1, before a sellout of 24,650 at Mapfre Stadium.

“Dos a Cero?” Meet “Dos a Uno.”

On an 89th-minute corner kick, Rafael Marquez, Mexico’s 37-year-old captain who played in the first Columbus showdown in 2001, sent a glancing header over substitute goalkeeper Brad Guzan to further numb an already shivering crowd.

It marked the Americans’ first defeat in a home qualifier since falling to Honduras 15 years ago at Washington’s RFK Stadium.

Given their history here, the Mexicans would not have been upset with draw. And given their first-half performance in Coach Jurgen Klinsmann’s flawed formation, the United States would not have rightly complained about salvaging a point in the first of 10 games en route to the 2018 World Cup in Russia. But on Miguel Layun’s late corner kick, no U.S. player marked Marquez.

Saddled with a defeat, the Americans will now have to quickly turn attention to Tuesday’s match at Costa Rica, which opened the hexagonal with a 2-0 victory at Trinidad and Tobago. They’ve never won a qualifier in Costa Rica (0-8-1).

Three of six teams in the final round of qualifying are assured of a World Cup berth, but two early defeats would increase the pressure.

“We have got to go down there and get a result, which we will do,” Klinsmann said. “Obviously Mexico is the biggest one we play. … It’s not a problem, but obviously it’s disappointing, no doubt about it.”

Klinsmann will almost certainly be without starting keeper Tim Howard, who strained a groin muscle in the first half. He will have an MRI exam Saturday.

Klinsmann will reflect on a night of riveting theater, on conceding Layun’s 20th-minute goal, on altering the game plan before halftime and on a stirring reply in the second half. Bobby Wood scored the equalizer in the 49th minute as the Americans reversed course and seized control.

But as the final whistle neared, they broke down on what the Americans take great pride in executing: a set piece.

“We were sleeping there,” midfielder Jermaine Jones said.

Two hours earlier, the crowd welcomed Mexico to the field for warm-ups with a cascade of “Dos a Cero” chants. The place was hopping. And the weather was freezing — just as the Americans had hoped. The game-time temperature was 43 degrees, but with a breeze, it felt colder.

No one in the stands seemed to mind, the electricity of the moment warming the rowdy confines.

Klinsmann sprung a few surprises in assembling his lineup. He played with three defenders instead of four, all center backs: Matt Besler, John Brooks and Omar Gonzalez.

The alignment heaped heavy responsibility on the wingers — Fabian Johnson and Timmy Chandler — to get up and down the flanks. Chandler’s only previous appearance this year came last month against Cuba. His terrific form with Bundesliga club Eintracht Frankfurt was too tempting for Klinsmann to overlook.

Mexico flooded Chandler’s side of the field, pinning him in and overwhelming the Americans with constant forays.

Christian Pulisic, an 18-year-old midfielder with electrifying skills, started in the heart of the attack. He had played in the previous qualifying round, in Copa America Centenario and in friendlies. Now he found himself in not only the lineup against Mexico, but in the playmaking role against the Americans’ greatest rivals in the most-anticipated World Cup qualifier of the four-year cycle.

Pulisic showed his talents, but Klinsmann was disappointed in Jones and captain Michael Bradley in deep-lying midfield roles. “The midfielders didn’t get into the one-against-one battles,” he said.

In the 10th minute, Jesus Manuel Corona forced Howard into a full-extension touch save that smacked the far post.

Ten minutes later, Howard was beaten. Giovani Dos Santos won the ball from Bradley 30 yards from the target, leaving it for Layun to drive low and hard. The ball caromed off Brooks and streaked beyond the goalkeeper’s reach.

Five minutes passed before Carlos Vela’s header kissed the crossbar.

Klinsmann’s formation had failed miserably. His team should have been down two or three goals. During an injury break, the team gathered near the sideline.

“Among us all,” Bradley said, “it was clear that it made sense to change.”

The new plan was a standard 4-4-2 arrangement. Chandler dropped to right back, Pulisic shifted to the left wing and Johnson flipped to the other flank. Pulisic’s golden chance in the box was foiled by a heavy touch. Mexico was unfazed by the American renewal, continuing to make dangerous runs in transition and forcing the hosts to retreat.

The Americans hit halftime trailing by just one goal — an undeserved lifeline.

They roared to life at the restart. The equalizer came on the counter. Brooks stepped up to intercept the ball and one-time an outlet pass to Jozy Altidore. The mighty striker made a tight turn on his marker and embarked on an assertive run.

He placed a through ball to Wood, whose deft first touch split two defenders inside the penalty area. His left-footed shot from 10 yards nicked a defender and glanced off goalkeeper Alfredo Talavera’s hand before rolling into the net for his eighth international goal.

The Americans could’ve gone ahead in the 58th minute when Gonzalez was left unmarked on Bradley’s corner kick, but the defender snapped his angled header at Talavera.

On even terms, the teams turned an open match into a grinding, physical affair, marred by confrontations and cards.

The game sprang back to life in the last 16 minutes. Wood tested Talavera with a wicked, rising shot. Pulisic revved his engines and was intentionally undercut by Carlos Salcedo, setting up a 25-yard free kick by Altidore that the keeper pushed over the bar.

As the match headed toward a draw, Marquez produced the stunning winner.

“It’s the first game,” Howard said. “We’ve got a whole bunch more to go.”