Rapid detection of GYPA, LDLR, HBGG, D7S8 and GC alleles by real-time fluorescence PCR

Abstract

We developed the allele-specific TaqMan polymerase chain reaction (AS-TaqMan PCR) and SYBR Green PCR assays for detecting glycophorin A (GYPA), low-density lipoprotein receptor (LDLR), hemoglobin G (HBGG), D7S8 and group-specific component (GC) alleles. We improved the specificity of detecting a nucleotide substitution by introducing the additional mismatches at position 2 (3 in GYPA). The differences between threshold cycle (Ct) values of different genotypes on each of the loci were statistically significantly different. All the genotypes agreed with the results using the AmpliType PM+DQA1 PCR Amplification and Typing kit. The AS-TaqMan PCR and SYBR Green PCR assays are simple, rapid, and accurate, as well as suitable for high-throughput applications in a forensic investigation.