Determination of ABO Variants Using ASP/SSCP Analysis
Jessica German · Cornerstone (Minnesota State University, Mankato) · 2010
The ABO blood system is the most commonly known of the blood type systems. It is composed of four different blood phenotypes: A, B, AB, and O. Each of these is made from two different genes. The different genes vary in just a few base pairs, and these differences are called single nucleotide polymorphisms (SNPs). Minor variants are not detected by normal blood typing, but they may be significant for the acceptance of blood transfusions or organ transplants. This study uses allele specific polymerase chain reaction (ASP) combined with single stranded conformation polymorphism (SSCP) to create gel patterns that distinguish among variant alleles. ASP is a method of amplifying DNA of a chosen sequence in the region containing one or more SNPs. SSCP uses denaturation, followed by quick cooling to create different foldings due to nucleotide changes. This combined method has successfully been used to detect common variants (A, B, O101, O201 and O303). These were verified by the development of a quantitative PCR (qPCR) SNP genotyping method. Sequencing will be used to determine the SNPs of the unique patterns representing the uncommon variants. These methods will allow us to screen for the ABO blood types variants by quick and inexpensive methods.