PCR for rapid detection of nine loci of Y-chromosomal DNA including SRY sequences in Turner syndrome patients

Detection of Y-chromosomal sequences is crucial for certain cases of phenotypic females since prophylactic gonadectomy is recommended to prevent germ cell tumor development in the future. This cross-sectional study aimed to develop a simple, rapid, and accurate method for determination of 9 loci of Y-chromosomal DNA spanning Yp, centromere, and Yq, including SRY sequences by using 2 polymerase chain reaction (PCR) conditions. The first PCR condition was used for Assay A, which yielded pentaplex PCR products consisting of loci ZFX/ZFY, SRY, AZFz, AZFb, and AZFc. The second PCR condition was used for Assay B, which yielded PCR products consisting loci PABY, AMGLXX/AMGLXY, Y-centromere, and YD. DNA from 20 Turner syndrome (TS) patients with different chromosomal aberrations were assessed. Normal female DNA and normal male DNA were used as controls. The pentaplex PCR together with 4 other simultaneous PCR reactions showed 10 positive PCR products of 9 loci in normal male controls and 3 mosaic TS patients (45,X/46,XY and 45,X/46,X,+marker). Five positive PCR products representing the regions PABY, ZFX/ZFY, and AMGLXX/AMGLXY and SRY were identified in 1 of 9 TS (monosomy; 45,X) patients. All female controls, 8 TS patients with monosomy X, and 8 mosaic TS patients with only X chromosome aberrations showed positive results for loci ZFX/ZFY and AMGLXX. The developed PCR conditions were demonstrated to accurately and rapidly detect 9 Y-chromosomal fragments including SRY sequences in TS patients.