SUMMARY: BACKGROUND: In a previous publication we described a method for Jk(a)/Jk(b), Fy(a)/Fy(b), S/s, K/k, Kp(a)/Kp(b), Js(a)/Js(b), Co(a)/Co(b), and Lu(a)/Lu(b) genotyping based on a microsphere suspension array. Here, an improved version of the assay is presented. METHODS: TWO MULTIPLEX POLYMERASE CHAIN REACTIONS (PCR) WERE DEVELOPED: one for amplification of samples routinely tested and the other for those systems that are tested less frequently.
View Article and Find Full Text PDFTransfus Med Hemother
May 2009
We have developed a high-throughput system for single nucleotide polymorphism (SNP) genotyping of alleles of diverse blood group systems exploiting Luminex technology. The method uses specific oligonucleotide probes coupled to a specific array of fluorescent microspheres and is designed for typing Jk(a)/Jk(b), Fy(a)/Fy(b), S/s, K/k, Kp(a)/Kp(b), Js(a)/Js(b), Co(a)/Co(b) and Lu(a)/Lu(b) alleles. Briefly, two multiplex PCR reactions (PCR I and PCR II) according to the laboratory specific needs are set up.
View Article and Find Full Text PDFWe describe the isolation and characterization of a novel HLA-A null allele, officially named A*0127N. The A*0127N exon 2, 3, and 4 nucleotide sequence is identical to that of A*010101 except at position 553, where a G is substituted by a T, resulting in a coding change in exon 3 (GAG>TAG) from Glu to the stop codon AMB. The mutation described is responsible for the premature ending of the translation.
View Article and Find Full Text PDFBackground: Traditionally, blood group typing has been performed with serologic techniques, the classical method being the hemagglutination test. Serotyping, however, may present important limitations such as scarce availability of rare antisera, typing of recently transfused patients, and those with a positive direct antiglobulin test. Consequently, serologic tests are being complemented with molecular methods.
View Article and Find Full Text PDFHum Immunol
December 2004
The major purpose of the present study was to investigate the frequency of human leukocyte antigen (HLA)-B27 alleles in healthy controls and in patients with ankylosing spondylitis (AS) and other HLA-B27-related diseases in the Greek Cypriot population. We selected 102 HLA-B27-positive individuals (60 controls and 42 patients). Typing of the HLA-B27 alleles was performed by polymerase chain reaction amplification with sequence-specific primers.
View Article and Find Full Text PDF