The Belgian's coach Reginald Willem had warned this morning it was unlikely Darcis would be able to play his second-round clash with Poland's Lukasz Kubot.
And the ATP, the governors of the men's tennis tour, confirmed a right shoulder problem had forced him out.
The 29-year-old caused the shock of the first round at the All England Club, knocking out 12-time grand slam winner Nadal in straight sets.
It emerged Darcis injured his shoulder during the match against Nadal, the biggest win of his career.
Coach Willems said the injury had left him unable to move his arm today.
Speaking before the official announcement arrived, Willems told Belgian publication Sudpresse: "There is a 99% chance that Steve will not play his second match.
"He has had an injection this morning but it will be very hard for him to go out to play.
"That's sport, unfortunately: the greatest victory of his career is followed by the greatest disappointment of his career."
Lukas Rosol, who beat Nadal in the second round last year, lost to Philipp Kohlschreiber in his next match in straight-sets.