Evaluation of Lewis genotyping by four PCR-based methods

Abstract: We developed PCR-based methods, confronting two pair primers (PCR-CTPP) and sequence-specific primers with a PCR-positive control (PCR-SSPPC) to analyze SNPs at nucleotide position 59 to reflect α-1, 4-fucosyltransferase (Le transferase) activity, which were analyzed by ABI PRISM® 3100 genetic analyzer. We compared 4 kinds of PCR-based methods, PCR restriction fragment-length polymorphism (PCR-RFLP), PCR sequence-specific-primers (PCR-SSP), PCR-CTPP and PCR-SSPPC. We found that all of these methods could determine Lewis genotyping correctly. The frequencies of Le and le alleles were 67.2% and 32.8% respectively. Both PCR-CTPP and PCR-SSPPC for Lewis genotyping are simple, reliable and applicable for forensic and clinical investigation. [Copyright &y& Elsevier]