High-throughput DNA typing of HLA-A, -B, -C, and -DRB1 loci by a PCR-SSOP-Luminex method in the Japanese population

被引:239
作者
Itoh, Y
Mizuki, N
Shimada, T
Azuma, F
Itakura, M
Kashiwase, K
Kikkawa, E
Kulski, JK
Satake, M
Inoko, H [1 ]
机构
[1] Tokai Univ, Sch Med, Dept Mol Life Sci, Div Mol Med Sci & Mol Med, Kanagawa 2591193, Japan
[2] Yokohama City Univ, Grad Sch Med, Dept Ophthalmol & Visual Sci, Kanazawa Ku, Kanagawa 2360004, Japan
[3] G&G Sci Co Ltd, Matsukawa, Fukushima 9601242, Japan
[4] Univ Tokushima, Inst Genome Res, Div Genet Informat, Tokushima 7708503, Japan
[5] Japanese Red Cross Tokyo Metropolitan Blood Ctr, Dept Lab, Minato Ku, Tokyo 1050011, Japan
关键词
HLA genotyping; PCR; SSOP; Luminex method; Japanese;
D O I
10.1007/s00251-005-0048-3
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
We have developed a new high-throughput, high-resolution genotyping method for the detection of alleles at the human leukocyte antigen (HLA)-A, -B, -C, and -DRB1 loci by combining polymerase chain reaction (PCR) and sequence-specific oligonucleotide probes (SSOPs) protocols with the Luminex 100 xMAP flow cytometry dual-laser system to quantitate fluorescently labeled oligonucleotides attached to color-coded microbeads. In order to detect the HLA alleles with a frequency of more than 0.1% in the Japanese population, we created 48 oligonucleotide probes for the HLA-A locus, 61 for HLA-B, 34 for HLA-C, and 51 for HLA-DRB1. The accuracy of the PCR-SSOP-Luminex method was determined by comparing it to the nucleotide sequencing method after subcloning into the plasmid vector using 150 multinational control samples obtained from the International HLA DNA Exchange University of California Los Angeles. In addition, we performed the PCR-SSOP-Luminex method for HLA allele typing on DNA samples collected from 1,018 Japanese volunteers. Overall, the genotyping method exhibited an accuracy of 85.91% for HLA-A, 85.03% for HLA-B, 97.32% for HLA-C, and 90.67% for HLA-DRB1 using 150 control samples, and 100% for HLA-A and -C, 99.90% for HLA-B, and 99.95% for HLA-DRB1 in 1,018 Japanese samples. The PCR-SSOP-Luminex method provides a simple, accurate, and rapid approach toward multiplex genotyping of HLA alleles to the four-digit or higher level of resolution in the Japanese population. It takes only approximately 5 h from DNA extraction to the definition of HLA four-digit alleles at the HLA-A, HLA-B, HLA-C, and HLA-DRB1 loci for 96 samples when handled by a single typist.
引用
收藏
页码:717 / 729
页数:13
相关论文
共 39 条
[1]   HLA-C genotyping in the Japanese population by the PCR-SSP method [J].
Ando, H ;
Mizuki, N ;
Ando, R ;
Miyata, Y ;
Miyata, S ;
Wakisaka, K ;
Inoko, H .
TISSUE ANTIGENS, 1996, 48 (01) :55-58
[2]   HLA-B40, B18, B27, and B37 allele discrimination using group-specific amplification and SSCP method [J].
Bannai, M ;
Tokunaga, K ;
Lin, L ;
Ogawa, A ;
Fujisawa, K ;
Juji, T .
HUMAN IMMUNOLOGY, 1996, 46 (02) :107-113
[3]   Graft-versus-host disease following living donor liver transplantation: high risk when the donor is HLA-homozygous [J].
Calmus, Y .
JOURNAL OF HEPATOLOGY, 2004, 41 (03) :505-507
[4]   INTERACTION BETWEEN DES-TYR1-GAMMA-ENDORPHIN AND HLA CLASS-I MOLECULES - SEROLOGICAL DETECTION OF AN HLA-A2 SUBTYPE [J].
CLAAS, FHJ ;
VANDERPOEL, JJ ;
CASTELLIVISSER, R ;
POOL, J ;
CHEN, RB ;
XU, KY ;
VANROOD, JJ .
IMMUNOGENETICS, 1985, 22 (04) :309-314
[5]   HLA class I and class II DNA typing and the origin of Basques [J].
Comas, D ;
Mateu, E ;
Calafell, F ;
Pérez-Lezaun, A ;
Bosch, E ;
Martínez-Arias, R ;
Bertranpetit, J .
TISSUE ANTIGENS, 1998, 51 (01) :30-40
[6]  
COMEY CT, 1993, J FORENSIC SCI, V38, P239
[7]   Quantitative, multiplexed detection of bacterial pathogens:: DNA and protein applications of the Luminex LabMAP™ system [J].
Dunbar, SA ;
Vander Zee, CA ;
Oliver, KG ;
Karem, KL ;
Jacobson, JW .
JOURNAL OF MICROBIOLOGICAL METHODS, 2003, 53 (02) :245-252
[8]  
Fulton RJ, 1997, CLIN CHEM, V43, P1749
[9]   GENE-FREQUENCIES AND HAPLOTYPIC ASSOCIATIONS WITHIN THE HLA REGION IN 916 UNRELATED JAPANESE INDIVIDUALS [J].
HASHIMOTO, M ;
KINOSHITA, T ;
YAMASAKI, M ;
TANAKA, H ;
IMANISHI, T ;
IHARA, H ;
ICHIKAWA, Y ;
FUKUNISHI, T .
TISSUE ANTIGENS, 1994, 44 (03) :166-173
[10]   HLA TYPING IN THE HONG-KONG CHEST-SERVICE BRITISH-MEDICAL-RESEARCH-COUNCIL STUDY OF FACTORS ASSOCIATED WITH THE BREAKDOWN TO ACTIVE TUBERCULOSIS OF INACTIVE PULMONARY-LESIONS [J].
HAWKINS, BR ;
HIGGINS, DA ;
CHAN, SL ;
LOWRIE, DB ;
MITCHISON, DA ;
GIRLING, DJ .
AMERICAN REVIEW OF RESPIRATORY DISEASE, 1988, 138 (06) :1616-1621