Serbia, third in Group E with three points, are one behind Brazil and Switzerland and need to beat the South Americans on Wednesday to qualify without depending on the result of the Switzerland-Costa Rica match which will be played at the same time.
International football is facing an uphill battle to compete with the riches of the club game, but Kolarov, who has won two English Premier League titles with Manchester City, said that nothing could match representing his country on the world stage.
"I'm getting towards the end of my career and this will be the most important game of my career," he told reporters.
"Players know what this is all about. This is a global stage, it's a once-in-a-career opportunity to shine.
"You always want to play the best in the world....you have to test yourself against the best and I am sure we will be prepared for that and will enjoy the moment."
There was also some fighting talk from Serbia coach Mladen Krstajic.
"We have no fear of Brazil, but we have to be patient, disciplined and play as a team," he said. "We have to satisfy the Serbian nation.
"We have shown we have the quality, we have a mix of seasoned players and young lads who will be playing for another 10 years for our national team."
"We have nothing to lose but if we are not ready to give our all, then we will have lost...You may lose, but you must not shirk your responsibility."
(Writing by Brian Homewood, editing by Neil Robinson)
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