Use it as the turning point in a short fail edit, after the setup and just before the consequence lands. Follow it with the Sad Trombone Sound Effect when the failure is complete, or start with the Nope Avi Sound Effect to signal doubt before the bad choice, then let this question close the sequence.
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This is a short, dry spoken reaction in which a single voice asks "what are you doing", with no music or other sound behind the words. The delivery is blunt and incredulous, giving listeners a flash of confusion, disbelief and mild panic. It works because the question sounds like a direct challenge to a baffling decision, catching the exact second someone realises a mistake has already been made.