With less than a fortnight to go before her title defence at the Australian Open, Williams confirmed she would not play in the United States’ final round-robin tie against the Czech Republic on Thursday.
The 34-year-old, who battled knee problems in the latter half of 2015, retired from her first singles match against Australian Jarmila Wolfe at the Hopman Cup in Perth on Tuesday.
She had already missed the United States’ opening Hopman Cup tie due to inflammation of her left knee before attempting to play against Wolfe.
The American retired when trailing 7-5, 2-1, having appeared extremely restricted in her movement during the match, particularly on her left side.
The injury cast significant doubt over her title defence in Melbourne, where she will aim for a seventh singles title later this month.
However, after the Wolfe match Williams said she was confident of being fully fit for Melbourne.
When informing Hopman Cup organisers of her withdrawal from the tournament, the Williams camp said they believed she would be fine after a couple of days’ rest.