The 16-year old Hawaiian carded a nine-over par 81 on Friday to miss the cut at the 84 Lumber Classic in Farmington, Pennsylvania.
The latest disappointment comes a week after Wie finished last in the European Masters in Switzerland but when asked whether she would reassess her ambition to play with men she said: "Nothing has changed.
"I'm not going to give up. I just had a bad two weeks, no more, no less.
"I have a clear idea of what I have to work on and what I have to do to get better.
"My score wasn't great. It was actually quite bad [but] I felt my game was 100 times better than last week, so I feel like I'm progressing, getting better, even if my score didn't show it."
Despite her upbeat assessment Wie was visibly crestfallen and she is doubtless coming to understand the huge gulf between the men's and women's games, particularly on longer courses where power matters so much.
She was one of the shortest players off the tee in Pennsylvania, averaging just 259 yards with her drives, which left her with many long and medium-iron second shots.
Her putting, not for the first time, let her down as she made just four of 14 putts in the five-to-10-foot range.
Wie, who turns 17 next month, is next scheduled to tee off with the boys in the Casio World Open, part of the Japan tour in November.
But many of her male colleagues believe a solid stint on the women's tour first would be a better strategy.
"I'm not going to give up. Everyone knows my ultimate goal is to play in The Masters"
Scott Verplank, a member of the US Ryder Cup team, told reporters: "Obviously she's some sort of phenomenon, being a 16-year-old girl who can play golf like she can but, honestly, there's not a male or female in this world who can compete out here at that age.
"If I was her adviser, I'd tell her to go kick all the ladies' tails around for four years and then if she wants to try again when she's 20 or 21, and grown up more and maybe a better player, come on back. She's certainly not scaring anybody around here."
Englishman Greg Owen agrees: "She's going to burn out if she's not careful. She needs to pick and choose more wisely and I don't think the PGA Tour is the place to do it."
But Wie remains defiant. "I'm not going to give up. Everyone knows my ultimate goal is to play in The Masters. My goal is to compete with the guys."
Elsewhere on Saturday, Shaun Micheel, the former US PGA champion and Englishman, Paul Casey advanced to the final of the World Match Play at Wentworth, England.
Micheel edged out Sweden's Robert Karlsson by finishing two up while Casey outplayed Colin Montgomerie to prevail six and five.