Karzai's seething words came after his meeting with families of the 16 Afghan civilians, including nine children and three women, killed in Kandahar province southern Afghanistan on Sunday.
"The Afghan government didn't receive cooperation from the USA regarding the surrender of the US soldiers to the Afghan government. This (civilian casualties) has been going on too long," Karzai told relatives of those killed, adding, "this behaviour cannot be tolerated."
The relatives insisted that there must have been more than one shooter and said they did not receive all the information from the Americans.
"This has been going on for too long. This is by all means the end of the rope here," Karzai told the media in Kabul. "This form of activity, this behavior, cannot be tolerated. It's past, past, past the time," Karzai said.
His comments came a day after a statement released by Karzai's office said that during his meeting with visiting US Defence Secretary Leon Panetta, the president "requested that the international forces come out of Afghan villages and stay in their bases" following the shootings in Kandahar.
Karzai said that the "Afghan security forces have the ability to provide security in the villages of our country," the statement said.
The fallout from the alleged killings has further strained tense US-Afghan relations.
Afghans have been demanding the soldier, who was flown out of Afghanistan by the US military, be returned to stand trial.
