Ch. Yang et al., HLA-DQB1 genotyping with simple automated DNA sequencing and single-strandconformation polymorphism analysis, J FORMOS ME, 99(9), 2000, pp. 698-703
Background and purpose: Accurate human leukocyte antigen (HLA) typing is im
portant for matching donors and recipients of bone marrow transplantation.
Because HLA is highly polymorphic, HLA genotyping is also a valuable tool i
n forensic identification of humans. The primary objective of this study wa
s to establish a simple, rapid, and economic HLA analysis system suitable f
or use in forensic applications.
Method: We used the primer pair DB130 and GH29 to amplify the HLA class II
DQB1 gene by polymerase chain reaction (PCR). The nucleotide sequences of t
he PCR products were analyzed with an ABI Prism 377 automatic DNA sequencer
, with the aid of its systematic analytical procedure. Some genotypes were
confirmed by single-strand conformation polymorphism (SSCP) analysis.
Results: We identified 15 alleles and 37 genotypes from 86 Taiwanese subjec
ts. The most frequent allele was 03011 (27.9%) and the most frequent genoty
pe was 03011/03011 (15.1%). Statistical analysis showed that the allelic di
versity was 0.862 and the power of discrimination was 0.948.
Conclusions: The results of this study indicate that the combined use of au
tomated sequencing with only one primer pair and SSCP provides a simple, ra
pid, and economic tool for analyzing the DQB1 gene. Compared with other seq
uencing methods that use a set of multiple primers, this method has two adv
antages. First, it is simpler and faster, because the HLAB1 genotype can be
determined in a single PCR reaction. Second, the use of only one primer se
t obviates the need for preferential annealing of any one primer set.