VALIDATION AND POPULATION STUDIES OF THE LOCI LDLR, GYPA, HBGG, D7S8,AND GC (PM LOCI), AND HLA-DQ-ALPHA USING A MULTIPLEX AMPLIFICATION AND TYPING PROCEDURE

Studies were performed to evaluate the forensic applicability of multi plex amplification of the loci low density lipoprotein receptor, glyco phorin A, hemoglobin G gammaglobin, D7S8, and group-specific component (PM loci) and simultaneous typing of these loci using a reverse dot b lot approach where allele specific oligonucleotide probes are immobili zed on a nylon membrane strip. These results were obtained by using th e AmpliType(R) PM PCR Amplification and Typing Kit. The experiments in cluded: mixed body fluid studies; chemical contaminant effects on the DNA in body fluid samples; the effect of typing DNA from body fluid sa mples deposited on various substrates; the effect of microorganism con tamination on typing DNA derived from blood and semen; the effect of s unlight and storage conditions on DNA typing; determination of the sen sitivity of detection of the PM test kit; determination of cross-react ivity of DNA from species other than human; typing DNA derived from va rious tissues from an individual; and an evaluation of the hybridizati on temperature of the assay. The data demonstrate that DNA exposed to a variety of environmental insults yields reliable PM typing results. Allele and genotype frequencies for six loci (PM loci and HLA-DQalpha) were determined in African Americans, Caucasians, southeastern Hispan ics, and southwestern Hispanics. All loci meet Hardy-Weinberg expectat ions and there is little evidence for association of alleles between t he loci. The frequency data can be used in forensic analyses and pater nity tests to estimate the frequency of a multiple locus DNA profile i n various general United States populations.