Mr Cameron said shadow chancellor Ed Balls should put the national interest first rather than questioning whether the £50 billion scheme should go ahead.
In a question to the Prime Minister, Tory MP Andrew Stephenson whose Pudsey constituency is in West Yorkshire, said: “Better rail connections to the south of England are... vital.
“Do you not agree with me it is absolutely outrageous for (Labour) to be challenging HS2 at the current time, putting in jeopardy jobs and investment in the north of England?”
Mr Cameron replied: “You are absolutely right to stand up for your constituents, to stand up for the north of England, because there is a real danger with Labour’s antics that they are letting down the north of England, letting down the Midlands.
“Let me remind the shadow chancellor what he said about these transport investments. He said this: ‘Nowhere is consensus more essential than on our transport infrastructure. Successive governments have ducked or delayed vital decisions on our national infrastructure, allowing short-term politics to get in the way’.
“By his own words, he is found guilty of short-termism, of petty politicking, rather than looking after the national interest.”
Concerned with the expense of the project, Labour appears to be cooling towards HS2 which will see the scheme’s first phase run from London to Birmingham in 2026, with a second Y-shaped route due to be completed to northern England in 2032/33.
Mr Balls has said it was vital that costs “stacked up” and there are fears that if Labour withdrew its support, the necessary legislation would not be able to make it through Parliament as there are a number of Tory MPs who are opposed to it.
Tomorrow, the High Speed Rail (Preparation) Bill - which will allow the Government to spend money planning the route in detail and buying up property from residents and businesses along the proposed track - will return to the House of Commons for report stage and third reading.
When the Bill was last in the Commons, a number of Tory MPs rebelled against the plans and Mr Cameron has said the scheme requires cross-party support to succeed.
In an exchange with Labour leader Ed Miliband earlier in the Commons, Mr Cameron added: “I tell you what is weak: too weak to stand up and admit their economic failures, too weak to stand up to Len McCluskey, who tried to wreck Scotland’s petrochemical industry. And too weak to stand up to the shadow chancellor (Ed Balls) on HS2.
“Let us just examine what happened with HS2 this week. The shadow chancellor touring the radio studios telling everyone it won’t go ahead, Labour local authority leaders begging you to stand up for this infrastructure scheme.
“What have you done? Cowered in your office, too weak to make a decision. To put it another way, Britain deserves better than this.”