ABO genotyping by polymerase chain reaction-restriction fragment length polymorphism

N. A. Mifsud, A. R. Haddad, J. A. Condon, R. L. Sparrow

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Abstract

Genotyping enables the identification of both maternally and paternally derived alleles. A number of protocols have been described for the genotyping of the ABO blood group system. Generally, these methods have a number of disadvantages including the use of hazardous reagents, being technically demanding, and the excessive use of materials. In this study, a relatively simple polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method is described. Four different amplifications were used that were specific for nucleotides sites 261, 526, 703, and 796 to distinguish the A, B, O 1 and O 2 alleles. The ABO genotypes of 294 random individuals were determined and were found to completely correlate with the serologic phenotypes. The protocol is applicable for investigations of weak or nonexpression of ABO alleles, paternity determinations, and population analysis. 143-148 ABO genotyping - identification of O 1, O(1*), and O 2 alleles using the polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) technique ABO polymorphism at the gene level has been investigated by molecular methods, predominantly sequencing and restriction fragment length polymorphism (RFLP). We describe the application of the polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) method, which is considered to be more versatile for large sample numbers, compared with conventional ABO genotyping by PCR-RFLP. PCR-SSO, while maintaining accurate and reliable results, reduces costs and labor. A population of 155 random individuals was investigated for the three O alleles, O 1 O(1*), and O 2. The allelic frequencies were 35 percent, 26 percent, and 5 percent, respectively. PCR-SSO results correlated completely with both serologic and PCR-RFLP results.

Original languageEnglish
Pages (from-to)143-148
Number of pages6
JournalImmunohematology
Volume12
Issue number4
Publication statusPublished - 1996
Externally publishedYes

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