The England captain and Manchester United striker has over 11.7m people subscribed to his posts on the social media platform.
Rooney's United team-mates Javier Hernandez (sixth with 5.57m followers), Juan Mata (seventh with 4.82m followers), David de Gea (eighth with 4.79m followers) and Victor Valdes (ninth with 4.16m followers) also made the top 10, while new signing Bastian Schweinsteiger is 16th with 2.59m followers.
Arsenal's Mesut Ozil is the second-most followed player with 9.79m followers, while Manchester City striker Sergio Aguero is third with 8.94m followers.
Radamel Falcao, who joined Chelsea on loan from Monaco this season after an unsuccessful season with Manchester United in 2014/15, is the fourth on the list with 8.92m followers, with Chelsea's Cesc Fabregas fifth with 7.93m followers.
A Manchester City contingent of David Silva (12th with 3.39m followers), Samir Nasri (14th with 2.6m followers), Vincent Kompany (17th with 2.26m followers) and Pablo Zabaleta (18th with 2.19m followers) dominated the 11-20 positions.
Out-of-favour Liverpool striker Mario Balotelli was the only Reds player to appear in the top 20, securing 11th position with 3.64m Twitter followers.
The numbers were taken on Monday, August 10, 2015.

Active and inactive

With 252 followers, West Ham and Switzerland U21 goalkeeper Raphael Spiegel registered the fewest followers out of all Premier League players with Twitter accounts.
Norwich City midfielder Nathan Redmond emerged as the most prolific Premier League Tweeter, posting more than 18,000 messages on the social media site.
Meanwhile, Chelsea youngster Ruben Loftus-Cheek has been the least active Premier League player on Twitter, posting just four messages.