A man has been arrested after Wycombe goalkeeper Ryan Allsop alleged homophobic abuse was aimed at him during Sunday's League One game at Tranmere.
The Merseyside club confirmed an incident did occur during the match at Prenton Park - which Wycombe won 2-0 to move to the top of the division - and is being dealt with by police.
It is understood that Allsop, 27, reported the abuse to the referee during the interval after he had kept goal at the Kop end in the first half.
A statement from the home club read: "Tranmere Rovers Football Club are very disappointed to learn of a homophobic comment made during today's match against Wycombe Wanderers.
"The club takes matters like this very seriously with a zero-tolerance policy.
"As a consequence of today's actions, one supporter has been arrested and the matter is being dealt with by the police."
The Football Association is also understood to be aware of the incident and will await the referee's report and seek further information from all parties involved before deciding how to proceed.
A further statement from Merseyside Police suggested referee John Busby had also been the target for homophobic abuse during the game, which was televised live.
Chief Inspector Jason Crellin said: "We're grateful to the player, both clubs and the stewarding staff for intervening and reporting these abhorrent incidents immediately.
"I also want to reassure them and everyone present and watching on television that we will be working alongside both clubs and the authorities to fully investigate the allegations.
"Hate crime has no place in our communities, and not least at a showcase sporting event attended and watched by many people."