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The Football Association says it is investigating an allegation that referee David Coote discussed giving a yellow card to a fan before a Championship match.
Coote, 42, says he denies the new claims and calls them "false and defamatory allegations."
The referee is already under investigation by the FA and has been suspended by the refereeing body the Professional Game Match Officials Limited (PGMOL) and European football's governing body Uefa, following separate allegations about his conduct.
A new report in the Sun newspaper on Tuesday claimed Coote exchanged messages with a fan discussing giving a yellow card before and after a game between Leeds and West Brom in 2019.
The exchange refers to a booking Leeds defender Ezgjan Alioski received in the match from Coote. There is no suggestion of any financial gain by Coote and the booking is regarded as entirely correct.
The contents of the report have not been verified by the BBC.
"I strongly refute these false and defamatory allegations," Coote said in a statement.
"Whatever issues I may have had in my personal life they have never affected my decision-making on the field.
"I have always held the integrity of the game in the highest regard, refereeing matches impartially and to the best of my ability."
Coote is one of the Premier League's most experienced officials and has been refereeing matches in the top flight since 2018.
The FA told the BBC: "These are very serious allegations and we are investigating as a matter of urgency."
The PGMOL said it takes a "zero-tolerance approach" to any breach of its code of conduct.
"The facts need to be established in light of these very serious allegations," it added.
"PGMOL board is committed to taking the appropriate action should any breach of that code be proven.
"David Coote remains suspended and subject to an ongoing disciplinary process by PGMOL, separate from the investigation into this matter, which will be carried out independently by the FA."

Leeds said they "are aware of the allegations" and added: "We respect and have full confidence in the FA, EFL and PGMOL regulations and processes. We will be making no further comment at this time."
Coote was initially suspended by the PGMOL on 11 November pending a full investigation after a video emerged that appears to show him make disparaging remarks about former Liverpool boss Jurgen Klopp. The FA started an investigation on 12 November.
On 13 November, the Sun published photos it says were taken during this summer's European Championship, alleging that they appear to show Coote sniffing a white powder through a rolled up US bank note.
Coote was working at the Euros in his capacity as a match official.
The PGMOL said it was taking those allegations "very seriously" and launched an investigation.
Uefa, European football's governing body, has also launched its own investigation and suspended Coote.
Neither the pictures nor the video have been verified independently by the BBC.
Last week, the PGMOL said Coote's welfare is "important to us" as its investigation into his conduct continues.
Former Premier League referee Mark Halsey told BBC Sport he was "stunned" by the new allegations against Coote and said they would be "damaging for all officials from grassroots all the way up", but he stressed the importance of waiting for the investigation to run its course.
Halsey added it was also important to look after Coote and believes mental health support needs to be put in place for referees.
He said: "I'm quite thick-skinned, I can quickly move on to the next game, but some can't and it is something that needs to be put in place, with all the officials at the top level."
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