TY - JOUR
T1 - Reference strand conformational analysis of untyped HLA class I molecules in two American Indian populations
AU - Cavett, J.
AU - Bennett, T.
AU - Sparkman, J.
AU - Kalve, I.
AU - Vornsand, J.
AU - Parlow, M.
AU - Warden, M.
AU - Leffell, Mary S.
AU - Zachary, Andrea A.
AU - Hildebrand, W.
PY - 2001
Y1 - 2001
N2 - Reference Strand Conformational Analysis (RSCA™) represents a high resolution Human Leukocyte Antigen (HLA) typing alternative. Our laboratory has routinely performed high throughput sequence-based typing (SBT), and we initially applied RSCA™ for resolving ambiguous cis/trans allelic combinations. Having found RSCA7 to be highly discriminative, we designed a study whereby SBT acts as a quality control for samples typed by RSCA™. We used RSCA™ to type a total of 352 samples from 2 different populations of American Indians at both the -A and the -B locus. Fifty samples from each population were also typed at the -A and the -B locus using SBT. Of the samples typed using SBT, 98% produced the types found using RSCA™. The RCSA results of one sample led to the discovery that it had been mistyped using SBT. Previously undetected alleles present in the sample pool were also discerned by RSCA™. One combination, A*2601 and A*2602, could not be distinguished with RSCA™ due to overlapping migration values: 1.4% of the samples produced this type and were resolved by SBT. In conclusion, RSCA™ blindly resolved 14 individual A alleles and 17 B alleles, including previously uncharacterized alleles. These results demonstrate that HLA typing by RSCA™ and SBT on one hardware platform provides an extremely precise HLA typing methodology.
AB - Reference Strand Conformational Analysis (RSCA™) represents a high resolution Human Leukocyte Antigen (HLA) typing alternative. Our laboratory has routinely performed high throughput sequence-based typing (SBT), and we initially applied RSCA™ for resolving ambiguous cis/trans allelic combinations. Having found RSCA7 to be highly discriminative, we designed a study whereby SBT acts as a quality control for samples typed by RSCA™. We used RSCA™ to type a total of 352 samples from 2 different populations of American Indians at both the -A and the -B locus. Fifty samples from each population were also typed at the -A and the -B locus using SBT. Of the samples typed using SBT, 98% produced the types found using RSCA™. The RCSA results of one sample led to the discovery that it had been mistyped using SBT. Previously undetected alleles present in the sample pool were also discerned by RSCA™. One combination, A*2601 and A*2602, could not be distinguished with RSCA™ due to overlapping migration values: 1.4% of the samples produced this type and were resolved by SBT. In conclusion, RSCA™ blindly resolved 14 individual A alleles and 17 B alleles, including previously uncharacterized alleles. These results demonstrate that HLA typing by RSCA™ and SBT on one hardware platform provides an extremely precise HLA typing methodology.
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M3 - Article
AN - SCOPUS:33749106701
SN - 0960-7420
VL - 28
SP - 225
JO - European Journal of Immunogenetics
JF - European Journal of Immunogenetics
IS - 2
ER -