Roethlisberger's lawyer, Ed Garland, confirmed the GBI's decision on Tuesday to The Associated Press.
"It is clear that they are doing a careful, thorough and extensive investigation," said Garland, who hoped the investigation would wrap up within a month.
Police are looking into the claim that Roethlisberger sexually assaulted a 20-year-old college student early March 5 at a nightclub in Milledgeville, Ga. Roethlisberger has not been charged, and Garland has disputed the claim and has hired his own team to investigate.
No reason was given as to why the GBI has withdrawn its request or when it did so. When asked what this means in terms of the status of the investigation of Roethlisberger, GBI public information officer John Bankhead declined to comment.
When asked how this would affect the timetable on when the GBI and the Milledgeville police department will complete their investigation and submit their case to the district attorney, so the DA's office can decide whether it will seek to press charges, Bankhead said: "I don't know. I'd have to have my crystal ball out but I don't have it with me."
When asked about the GBI withdrawal of the DNA request and what that meant to the case, Lee Parks, an attorney for the accuser, said: "We're not commenting on the investigation. It's not our place to do that."
Roethlisberger, who owns a home about 30 miles north of Milledgeville on Lake Oconee, was celebrating his 28th birthday with friends at the club and other spots around the college town.
NFL commissioner Roger Goodell said Monday he planned to meet with Roethlisberger about the star's off-field problems. The quarterback's legal team said it wouldn't object to the meeting.
Steelers president Art Rooney II has said the team is concerned and closely monitoring the Milledgeville situation.
Roethlisberger skipped voluntary workouts with the team last week. Steelers coach Mike Tomlin said he has no doubt the quarterback will attend all required offseason practices and training sessions. Those begin March 29 for the regulars.
"When it's time for him to be there, he'll want to be there," Tomlin said. "He doesn't mind toting the burden of being the guy carrying our football team."
Information from The Associated Press was used in this report.