A rapid and quantitative DNA sex test: fluorescence-based PCR analysis of X-Y homologous gene amelogenin.

A rapid, simple and reliable sex test that entails PCR amplification of a segment of the X-Y homologous gene amelogenin has been developed. We used a single pair of primers spanning part of the first intron which generated 106-bp and 112-bp PCR products from the X and Y homologues, respectively, that can be analyzed simply by agarose gel electrophoresis. Less than 1 ng of template DNA is required for gender assignment, and the test has been automated by the fluorescent tagging of the PCR products that are then quantitated during electrophoresis by automated fluorescence-detection technology. Quantitation enables sex chromosome aneuploidy to be determined, and the amelogenin intron sequence can also be co-amplified with several highly polymorphic microsatellite loci, thereby providing a combined gender/identity DNA test.