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Notes -
There is a non-invasive test for having a Y chromosome (you just need a DNA sample, and before DNA testing there was a histological test for the presence of multiple X chromosomes which could be done with a cheek swab). The IOC used karyotype testing from 1958 to 1999 - it was abandoned after a small number of false positives of which the famous ones were cases of Complete Androgen Insensitivity Syndrome - CAIS women have XY chromosomes and testicles which produce testosterone, but their body doesn't respond to the testosterone so they develop female external genitalia in utero and female physiology after puberty. Everyone agreed that in principle CAIS women should be allowed to compete as women because they don't have an unfair advantage, and people who opposed sex testing used this as a lever.
But the practical protocol of "include a karyotype test in the standard battery of medical tests that elite athletes undergo, and discretely make further enquiries about XY women" works if you let it. The problem is that you have to adjudicate borderline cases like Caster Semeneya, and in the rare genuine intersex cases people will hate you for good reasons whichever way you rule. If you say "no gatekeeping - anyone AFAB can compete as a woman" then you are giving the competition away to countries willing to modify birth certificates.
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