ABO blood group genotyping by quenching probe method.
We developed a multiplex ABO genotyping method with quenching probes (Q-probe). In this method, it is possible to discriminate the mutations, not only frequently used positions 261 and 796 but also position 703 in a single PCR. Each probe was designed to have cytosine residue at 5' or 3' end and labeled with three different fluorescence dyes, enabling the triplex detections of these polymorphisms. All polymorphisms were successfully detected by using fluorescence labeled Q-probe in a specifically amplified PCR product. Each Q-probe showed unique dissociation patterns depending on the polymorphism types. All of the results obtained with Q-probe were compared with standard serotyping and TaqMan PCR method and resulted in complete match with each other. Consequently, these results indicated that multiplex ABO genotyping method is quite accurate and convenient method for the determination of ABO genotype.
Nihon University School of Dentistry, Department of Legal Medicine and Dental Research Center, Division of Social Dentistry, 1-8-13 Kanda Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
Hirofumi
H
Tsutsumi
Masatake
M
Asano
Yoshiyuki
Y
Hagiwara
Hiroaki
H
Nogami
Hikaru
H
Izawa
Sayaka
S
Maruyama
Toshinobu
T
Komuro
D000017:ABO Blood-Group System / Q000235:genetics*; D003596:Cytosine / Q000737:chemistry; D005456:Fluorescent Dyes; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D015335:Molecular Probes / Q000737:chemistry / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
D000017:ABO Blood-Group System; D005456:Fluorescent Dyes; D015335:Molecular Probes; D003596:Cytosine