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Blood, Vol. 95 No. 4 (February 15), 2000:
pp. 1487-1492
Single-tube multiplex PCR-SSCP analysis distinguishes 7 common
ABO alleles and readily identifies new alleles
Shea Ping Yip
From the Department of Nursing and Health Sciences, the Hong Kong
Polytechnic University, Hong Kong SAR, China.
The ABO blood group is clinically the most important blood group
system. Elucidation of the molecular basis of the ABO polymorphism allows genotype determination without family studies. Described here is
a new method based on the simultaneous amplification by polymerase
chain reaction (PCR) of 3 fragments from exon 6, and 5' and
3' ends of exon 7 of the ABO gene, followed by single-strand conformation polymorphism (SSCP) analysis. This multiplex PCR-SSCP protocol allows the well-established base changes at 9 nucleotide positions 261, 297, 467, 526, 646, 657, 681, 1059, and 1096 to be
assayed simultaneously so that 7 common alleles (A1,
A1v, A2, B, O1,
O1v, and O2) can be distinguished in a
single-tube single-lane format. Each allele was characterized by a set
of 3 haplotype-specific SSCP patterns. Chinese (n = 125) and white
European (n = 98) samples were analyzed, and their
genotypes were found consistent with the serologic phenotypes or could
be deduced unambiguously. Fifteen samples (2 Chinese and 13 white
European) were each found carrying at least 1 rare allele. Most
of these alleles were new and some might be generated by intragenic
recombination. This technique is the simplest, quickest, and most
informative method reported to date and also readily identifies new alleles.
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