"Our very weak and ineffective leader Paul Ryan had a bad conference call where his members went wild at his disloyalty," Trump, 70, said in an apparent angry tweet against the top party leader.
The tweet from Trump comes a day after House of Representatives Speaker Ryan told his party leaders in an emergency conference call that he would focus on maintaining majority in the Congress and would not be supporting his presidential nominee.
"Despite winning the second debate in a landslide (every poll), it is hard to do well when Paul Ryan and others give zero support!," the real estate billionaire from New York, who has 12.3 million followers on Twitter, said.
Latest polls indicated that Trump is trailing behind his Democratic rival Hillary Clinton, 68, by double digit.
After the surfacing of Trump's remarks making lewd comments about women, a large number of Republican leadership has deserted him. More than two dozens of the party leadership has asked him to withdraw from the race.
Soon after Ryan's conference call with his party leaders, Trump tweeted that Ryan is wasting his time.
"Paul Ryan should spend more time on balancing the budget, jobs and illegal immigration and not waste his time on fighting Republican nominee," he tweeted a day earlier.
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