Wayne Rooney is prepared to take on Manchester United and issue a formal transfer request to try to force the club to change their mind and allow him the move he wants to Chelsea.

Rooney is increasingly thinking he will have to take the initiative after United revealed they had turned down a second bid from Stamford Bridge – an initial £25m with another £5m in potential add-ons – and categorically stated that the striker would not get his wish. "He is not for sale," a club spokesman said.

The Premier League champions insist their stance will not change, that this is not merely a bargaining technique and that Rooney needs to buckle down and start preparing for a World Cup season.

That was followed by Rooney pulling out of the pre-season fixture against AIK in Stockholm on Tuesday, the official explanation being that he has a shoulder injury. Rooney, who had returned early from the club's summer tour after citing a hamstring strain, has not played since the end of last season and is a major doubt for England's game against Scotland at Wembley on Wednesday week.

By then, there is a strong possibility that he will have formalised his position at Old Trafford with a transfer request. The England international, intrigued by the possibility of working for José Mourinho and disillusioned with life at Old Trafford, will cite long-standing and deep-rooted unhappiness because of his fractured relationship with Sir Alex Ferguson and, if necessary, make it clear his stance has not altered because of the change of manager.

Rooney believes Ferguson will still play a considerable role in the David Moyes era. The 27-year-old is nursing a grudge against the man who brought him to the club – other players have reported being shocked by the friction between the two last season – and Rooney has made up his mind that he wants no part of it regardless of the fact that he is well positioned, on 197 goals, to overtake Sir Bobby Charlton, with 247, as the leading scorer in United's history.

That leaves Moyes in the invidious position of knowing that one of United's category-A players does not want to be at Old Trafford and is not interested in any attempts to talk him around or the possibility of a testimonial in a year's time. Chelsea are the only club to have bid for Rooney and United, far more minded to sell him to a foreign club, believe it would be a critical blow in the title race if they allowed a player of his ability to join one of their major domestic rivals.

Moyes is also acutely aware that, with no Rooney, United would be light on attackers, particularly bearing in mind Robin van Persie's past injury problems. Javier Hernández and Daniel Welbeck would provide other options but United's concerns are obvious and balanced against the possibility of leaving Mourinho with strength in depth, in the form of Rooney, Demba Ba, Fernando Torres and Romelu Lukaku.

United have already found it a difficult summer in terms of transfer business and, with two unsuccessful bids already, clearly have their work cut out persuading Barcelona to sell Cesc Fábregas.

They are, however, pressing ahead. A third bid in the region of £35m is being put together despite Barcelona's sporting director, Andoni Zubizarreta, claiming at the weekend that United had accepted the deal would not happen and Arsène Wenger's observation, after his own inquiries on behalf of Arsenal, that a bid would be a futile exercise. Wenger's information is that his former player will stay at Camp Nou for at least another year but the United chief executive, Ed Woodward, has been given the encouragement to believe a move is still possible and is keen to support Moyes in the transfer market.

While Manchester City have spent over £97m on Stevan Jovetic, Alvaro Negredo, Fernandinho and Jesús Navas, Woodward and his colleagues recognise losing Rooney without a like-for-like replacement lined up would be a significant setback to Moyes.

Rooney played in a practice match against Real Betis at United's training ground on Saturday and has been pencilled in to feature in Rio Ferdinand's testimonial match at Old Trafford on Friday. That is now in doubt, as is any participation in the Community Shield against Wigan Athletic two days later, which would at least spare him the possibility of a hostile reception from the 40,000-plus United supporters at Wembley.

"Wayne hurt his shoulder in a fall during a match on Saturday," Moyes said. "It's not too serious and I don't expect it to keep him out for too long. I'm disappointed he's missing the game because we were keen to get him back involved and ready to play."

Manchester United squad

Lindegaard, Amos; Rafael da Silva, Evans, Evra, Jones, Smalling, Vidic; Anderson, Bébé, Carrick, Giggs, Nani, Zaha; Henríquez, Kagawa, Van Persie, Welbeck.