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Halep had been confident of opening her title defence on Centre Court on Tuesday, but revealed on Friday morning that she faces a further spell out with the issue, which initially forced her to withdraw from the French Open.
The Romanian said: “It is with great sadness that I’m announcing my withdrawal from the Championships as my calf injury has not fully recovered.
“I gave it everything I had in order to be ready to play Wimbledon and, after having such special memories from two years ago, I was excited and honoured to step back on these beautiful courts as defending champion. Unfortunately, my body didn’t cooperate and I’ll have to save that feeling for next year.
“I can honestly say that I’m really down and upset about having to take this decision. This period has been difficult but to miss the last two Majors has made it even more challenging mentally and physically.
“We will see what the future holds but I’m hoping it will make me a stronger person and athlete.”
Halep’s withdrawal leaves the women’s draw without two of its biggest names after Naomi Osaka had previously pulled out because of her ongoing mental health issues.