Detection of MNS hybrid molecules in the Thai population using PCR-SSP technique

We developed a polymerase chain reaction-sequence-specific primer (PCR-SSP) technique to screen for hybrid molecules in the MNS blood group in the Thai population using two sets of newly designed primers specific for four GYP(B-A-B) hybrids, GP.Mur, GP.Hop, GP.Bun and GP.HF, and two GYP(A-B-A) hybrids, GP.Vw and GP.Hut. One thousand and forty-one blood samples were tested with human anti-Mi(a) by conventional tube technique, and 598 samples of these were tested by the PCR-SSP technique. Ninety-four samples (9.03%) were strongly positive with human antisera by conventional tube technique. For PCR-SSP test results, the GP.Hut, GP.Mur, GP.Hop, GP.Bun and GP.HF genotypes were amplified with the first set of primers, whereas GP.Vw genotype was amplified with a second set of primers. The GYP(A-B) hybrids (GP. Hil and GP.JL), GYP(A-B-A) hybrids (GP.Nob, GP.Joh and GP.Dane), GYPA, GYPB and GYPE were not amplified by either set of primers. Results of testing 94 Mi(a+) and 504 Mi(a-) by conventional tube technique and PCR-SSP were concordant. This study shows that analysis by PCR-SSP is simple and convenient; therefore, it can be used as an alternative to conventional tube technique for mass screening for MNS hybrids, especially when specific antisera are not available.