The 29-year-old started and provided two assists as the Three Lions secured an emphatic 6-2 victory over Iran in their opening Group B fixture at the 2022 World Cup on Monday.
However, Kane was withdrawn in the final 15 minutes following a lunging challenging from Iran's Morteza Pouraliganji, with Callum Wilson brought on as his replacement.
England's No.9 was then spotted limping after the match with strapping wrapped round his ankle as he left the Khalifa International Stadium in Doha.
Three Lions manager Gareth Southgate played down suggestions that Kane had suffered a serious injury when he spoke at a press conference after the game.
"I think [Kane is] fine. It looked like a bad tackle but he carried on in the game. We took him off because we felt it was a moment in the game we could do that," Southgate told reporters.
"Really pleased with our attacking play. We know that Iran are usually very difficult to score goals against, so it's a credit to our players, the way their movement, the quality of our passes, the quality of our finishing.
"I didn't like the end of the game. To concede two goals the way we did, isn't the level we need. So we're going to have to be better than the level we were today in certain aspects of our game against the USA."
According to The Athletic, Kane will undergo a scan on his ankle before England's second Group B clash against USA on Friday.
Kane, who has been frustrated by ankle issues in the past, won the World Cup Golden Boot at the 2018 tournament in Russia after netting six goals for England on their way to the semi-finals.
The 76-cap international has scored 51 times in total for his country and he is just three goals away from surpassing Wayne Rooney as England's all-time leading scorer.
Kane is not the only fitness concern within Southgate's camp, with Manchester United centre-back Harry Maguire hoping to recover from illness - which forced him off in the closing stages against Iran - ahead of Friday's clash with USA.
Meanwhile, Leicester City's James Maddison has missed training again due to a knee problem, but Manchester City's Kyle Walker has taken part in first-team training and he is pushing to be involved against USA.