A COMPARATIVE-STUDY OF HLA-DRB1 TYPING BY STANDARD SEROLOGY AND HYBRIDIZATION OF NONRADIOACTIVE SEQUENCE-SPECIFIC OLIGONUCLEOTIDE PROBES TO PCR-AMPLIFIED DNA

被引：53

作者：

MICKELSON, E ^{[1
]}

SMITH, A ^{[1
]}

MCKINNEY, S ^{[1
]}

ANDERSON, G ^{[1
]}

HANSEN, JA ^{[1
]}

机构：

[1] UNIV WASHINGTON,SCH MED,DEPT MED,SEATTLE,WA 98195

来源：

TISSUE ANTIGENS | 1993年 / 41卷 / 02期

关键词：

HLA-DR SEROLOGY; OLIGOTYPING; POLYMERASE CHAIN REACTION;

D O I：

10.1111/j.1399-0039.1993.tb01984.x

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

A double-blind study was carried out to evaluate the relative performance and reliability of the PCR/SSOP assay compared to conventional serological typing in identifying HLA-DR alleles. A total of 268 consecutive samples were entered into the study. In 14 (5.2%) of the cases, HLA-DR serology could not be performed due to poor cell viability, while in seven (2.6%) of the cases, PCR/SSOP typing could not be performed due to poor amplification or to contamination with exogenous DNA. Among samples that were successfully typed by both methods, serologic typing correctly identified 455/465 (97.9%) DR antigens, while PCR/SSOP correctly identified 464/465 (99.8%) DR alleles (p=0.0117, McNemar's test). The majority of discrepancies in serologic typing resulted from a lack of discriminative alloantisera to identify DR6 or DR103. For the overall sample set (N=268), serology provided accurate results in 244 (91.0%) cases, while PCR/SSOP provided accurate results in 260 (97.0%) cases (p=0.0037). The results of this study demonstrate that PCR/SSOP typing for HLA-DRB1 alleles provides results that are equal to or surpass serological typing for HLA-DR antigens. In addition, the PCR/SSOP approach offers the advantages of better reagent availability, lower cost, more rapid turn-around time, and greater accuracy, all of which would warrant its use as an HLA typing method of choice.

引用

页码：86 / 93

页数：8

共 19 条

[11]

MARCADET A, 1989, IMMUNOBIOLOGY HLA, V1, P553

[12]

MICKELSON EM, 1991, MARROW TRANSPLANTATI, V1, P8

[13]

PETERSDORF EW, 1991, IMMUNOGENETICS, V33, P267

[14] POLYMORPHISM OF HLA-DRW52-ASSOCIATED DRB1 GENES AS DEFINED BY SEQUENCE-SPECIFIC OLIGONUCLEOTIDE PROBE HYBRIDIZATION AND SEQUENCING [J].

PETERSDORF, EW ;

SMITH, AG ;

HAASE, AM ;

MARTIN, PJ ;

HANSEN, JA .

TISSUE ANTIGENS, 1991, 38 (04) :169-177

[15] PRIMER-DIRECTED ENZYMATIC AMPLIFICATION OF DNA WITH A THERMOSTABLE DNA-POLYMERASE [J].

SAIKI, RK ;

GELFAND, DH ;

STOFFEL, S ;

SCHARF, SJ ;

HIGUCHI, R ;

HORN, GT ;

MULLIS, KB ;

ERLICH, HA .

SCIENCE, 1988, 239 (4839) :487-491

[16]

SASAZUKI T, IN PRESS HISTOCOMPAT

[17]

VANLEEUWEN A, 1990, ASHI LABORATORY MANU, P201

[18] THE APPLICATION OF OLIGONUCLEOTIDE PROBES TO HLA CLASS-II TYPING OF THE DRB SUB-REGION [J].

VAUGHAN, RW ;

LANCHBURY, JSS ;

MARSH, SGE ;

HALL, MA ;

BODMER, JG ;

WELSH, KI .

TISSUE ANTIGENS, 1990, 36 (04) :149-155

[19]

WORDSWORTH BP, 1990, IMMUNOGENETICS, V32, P413

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