FY*X real‐time polymerase chain reaction with melting curve analysis associated with a complete one‐step real‐time FY genotyping

Abstract

The real-time technology method is rapid and accurate for FY genotyping. From now, we are able to detect the FY*X allele in all the alleles combinations studied. Regarding its significant frequency, the detection of the FY*X allele is useful for the correct typing of blood donors and recipients considering the therapeutic use of blood units and the preparation of test red blood cells for antibody screening.

Future of Molecular Testing for Red Blood Cell Antigens

“…Detection of red cell blood group SNPs by melting-curve analysis has been used to genotype for KEL1/2, JK1/2, FY1/ 2, FY0, FYX, MNS1-4, DO1/2, CO1/2, LU1/2, YTA/B, and DIA/DIB. [6][7][8][9] Pyrosequencing is a method of DNA sequencing based on the sequencing-bysynthesis principle. It differs from the common Sanger sequencing relying on the detection of pyrophosphate release on nucleotide incorporation, rather than chain termination with dideoxynucleotides.…”

Blood group genotyping by high‐throughput DNA analysis applied to 356 reagent red blood cell samples

“…Real‐time PCR with FRET method (LightCycler 2.0 and 1.2 Roche Diagnostics GmbH, Mannheim, Germany) was used to analyze the three polymorphisms FY * A/FY * B , FY * X/nonFY * X , and FY * 0/non‐FY * 0 ( FY * Fy/nonFY * Fy ). The primer and hybridization probe design as well as the PCR conditions were as described by Araujo and coworkers 6 and Ansart‐Pirenne and coworkers 5 . The genotyping of KEL * 1/KEL * 2 polymorphism by real‐time PCR on the LightCycler was described in 2002 by Araujo and coworkers 7 .…”

“…The main genetic event responsible for blood group polymorphisms is a single-nucleotide polymorphism 2-4 that can be analyzed by different polymerase chain reaction (PCR) types. [5][6][7][8][9][10][11] PCR-restriction fragment length polymorphism and allele-specific PCR were the first methods used for DNA analysis of blood group systems; real-time PCR is at present a commonly used method. More recently high-throughput genotyping systems have been developed to allow rapid determination of major and minor blood group polymorphisms.…”

Duffy Blood Group System

“…If separate tests are employed for the two SNPs, the results can be interpreted as shown in Table 8.5 and will give a high level of accuracy, though a false result will occur if the rare allele with − 67C and 125G is present. Further tests of the SNP at position 265 can be added to detect the presence of FY * X [34,39,47,48,120] . Further tests of the SNP at position 265 can be added to detect the presence of FY * X [34,39,47,48,120] .…”