HLA TYPING FOR DR3 AND DR4 USING ARTIFICIAL RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM PCR FROM ARCHIVAL DNA

被引:8
作者
HORTON, VA
BUNCE, M
DAVIES, DR
TURNER, RC
LO, YMD
机构
[1] CHURCHILL HOSP,OXFORD TRANSPLANT CTR,OXFORD OX3 7LJ,ENGLAND
[2] JOHN RADCLIFFE HOSP,DEPT CELLULAR PATHOL,OXFORD OX3 9DU,ENGLAND
[3] JOHN RADCLIFFE HOSP,NUFFIELD DEPT CLIN MED,OXFORD OX3 9DU,ENGLAND
基金
英国惠康基金;
关键词
HLA TYPING; ARTIFICIAL RFLP PCR; DIABETES;
D O I
10.1136/jcp.48.1.33
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
Aim - To develop polymerase chain reaction based artificial restriction fragment length polymorphism (artificial RFLP PCR) assays for DR3 and DR4 alleles of the multiallelic DRB1 locus and to apply them to paraffin wax embedded archival material. Methods - Sixty five samples from DRB1 typed cell lines were analysed using the artificial RFLP PCR method to determine the specificity and sensitivity of the system. Results - The artificial RFLP method for typing the DRB1 showed 100% accuracy in the 65 samples previously typed using allele specific PCR and serology. The samples included 18 combinations of alleles that included DR3, 18 that included DR4, four that were DR3/DR4 heterozygotes, and 10 samples that were neither DR3 nor DR4. Typing of 10 paraffin wax embedded samples using artificial RFLP PCR was in complete agreement with previous typing at the DRB1 locus. Conclusion - The application of artificial RFLP PCR for the analysis of multiallelic loci, such as those of the HLA system, in archival DNA samples achieved. Artificial RFLP robust, easily implemented, non-isotopic system and may be useful for large retrospective studies.
引用
收藏
页码:33 / 36
页数:4
相关论文
共 19 条
  • [1] GENERATION OF DIGOXIGENIN-LABELED DOUBLE-STRANDED AND SINGLE-STRANDED PROBES USING THE POLYMERASE CHAIN-REACTION
    AN, SF
    FRANKLIN, D
    FLEMING, KA
    [J]. MOLECULAR AND CELLULAR PROBES, 1992, 6 (03) : 193 - 200
  • [2] RAPID HLA-DRB1 GENOTYPING BY NESTED PCR-AMPLIFICATION
    BEIN, G
    GLASER, R
    KIRCHNER, H
    [J]. TISSUE ANTIGENS, 1992, 39 (02): : 68 - 73
  • [3] A DNA-RFLP TYPING SYSTEM THAT POSITIVELY IDENTIFIES SEROLOGICALLY WELL-DEFINED AND ILL-DEFINED HLA-DR AND DQ ALLELES, INCLUDING DRW10
    BIDWELL, JL
    BIDWELL, EA
    SAVAGE, DA
    MIDDLETON, D
    KLOUDA, PT
    BRADLEY, BA
    [J]. TRANSPLANTATION, 1988, 45 (03) : 640 - 646
  • [4] RAPID HLA-DQB TYPING BY 8 POLYMERASE CHAIN-REACTION AMPLIFICATIONS WITH SEQUENCE-SPECIFIC PRIMERS (PCR-SSP)
    BUNCE, M
    TAYLOR, CJ
    WELSH, KI
    [J]. HUMAN IMMUNOLOGY, 1993, 37 (04) : 201 - 206
  • [6] DNA TYPING OF HLA CLASS-II GENES IN B-LYMPHOBLASTOID CELL-LINES HOMOZYGOUS FOR HLA
    KIMURA, A
    DONG, RP
    HARADA, H
    SASAZUKI, T
    [J]. TISSUE ANTIGENS, 1992, 40 (01): : 5 - 12
  • [7] DETECTION OF HEPATITIS-B PRECORE MUTANT BY ALLELE SPECIFIC POLYMERASE CHAIN-REACTION
    LO, ESF
    LO, YMD
    TSE, CH
    FLEMING, KA
    [J]. JOURNAL OF CLINICAL PATHOLOGY, 1992, 45 (08) : 689 - 692
  • [8] INVITRO AMPLIFICATION OF HEPATITIS-B VIRUS SEQUENCES FROM LIVER-TUMOR DNA AND FROM PARAFFIN WAX EMBEDDED TISSUES USING THE POLYMERASE CHAIN-REACTION
    LO, YM
    MEHAL, WZ
    FLEMING, KA
    [J]. JOURNAL OF CLINICAL PATHOLOGY, 1989, 42 (08) : 840 - 846
  • [9] GEOGRAPHICAL VARIATION IN PREVALENCE OF HEPATITIS-B VIRUS-DNA IN HBSAG NEGATIVE PATIENTS
    LO, YMD
    LO, ESF
    MEHAL, WZ
    SAMPIETRO, M
    FIORELLI, G
    RONCHI, G
    TSE, CH
    FLEMING, KA
    [J]. JOURNAL OF CLINICAL PATHOLOGY, 1993, 46 (04) : 304 - 308
  • [10] HLA TYPING BY DOUBLE ARMS
    LO, YMD
    MEHAL, WZ
    WORDSWORTH, BP
    CHAPMAN, RW
    FLEMING, KA
    BELL, JI
    WAINSCOAT, JS
    [J]. LANCET, 1991, 338 (8758) : 65 - 66