摘要:

Abstract:We have studied restriction fragment length polymorphism (RFLP) in the region 300 kb centromeric to the HLA‐B locus. Four probes were used: one was genomic DNA derived from the tumor‐necrosis factor (TNF)‐β gene, one was a cDNA for the BAT3 gene, and two single‐copy genomic probes, R5A and M20A. The order of these markers from HLA‐B towards the centromere is M20A, R5A, TNF and BAT3. The BAT3 and TNF‐β probes each detected two allelic bands with Taq I and Nco I digestion, respectively; the R5A and M20A probes each detected three polymorphic allelic bands with BstEII digestion. To determine if these restriction polymorphisms are preferentially associated with certain HLA‐B and ‐DR haplotypes, a total of 153 HLA haplotypes was analyzed. The haplotypes Al, B8, DR3 and A3, B7, DR2 were each associated with a distinct combination of polymorphisms identified at these four sites, thereby demonstrating that the strong linkage disequilibrium characteristic of these haplotypes extends also to this segment of the class III region. In contrast, haplotypes that are not in positive linkage disequilibrium, such as A1,B8,DR4 and A2,B7,DR3, showed ho preferential association with any of these polymorphisms. The antigens HLA‐B27 and B35 were also found to be in positive linkage disequilibrium with RFLP patterns at three of these sites, and HLA‐B14,B35,B44,Bw57 and Bw62 were found preferentially associated with polymorphisms at one or two of these sites, independent of the DR antigen present. These data further demonstrate that genetic linkage disequilibrium in the HLA class III region is complex and variable among dif

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01871.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Abstract:The two DR1‐associated cellular specificities Dwl and Dw20, as well as DR'Br’(Dw'BON'), cannot be unequivocally assigned by serological typing or restriction fragment length polymorphism (RFLP) analysis. We have developed and compared two polymerase chain reaction‐based (PCR) typing methods for distinguishing these DRB1 alleles; allele‐specific amplification of DRB1*01 alleles followed by an agarose gel electrophoresis detection step and group‐specific DRBI*01 amplification followed by hybridization with sequence‐specific oligonucleotide probes. The two typing strategies gave completely concordant results in the 33 DRB1*01‐positive and the 46 DRB1*01‐negative individuals and cell lines studied. No false‐negative or false‐positive typing results were obtained. All possible heterozygous combinations of the DRB1*0101‐0103 alleles could be distinguished by both typing methods. DRB1*01 subtyping by allele‐specific PCR amplification was performed in less than 3 hours, including PCR amplification, detection and interpretation steps. The technique will be a valuable complement to DR typing by serology and RFLP analysis. Allele‐specific DRB1 amplifications or group‐specific amplifications followed by directed allele‐specific amplifications of DRB1 alleles, typing based on the absence or presence of amplified products, may well prove to be the technical innovation that will firmly establish PCR‐based DR typing in

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01872.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Abstract:Locus HLA‐DRB3 codes for the serologically defined supertypic specificity DRw52 in HLA‐DR3, ‐5 and ‐w6 haplotypes. Three specificities of DRw52 (DRw52a, ‐b and ‐c) can further be distinguished by cellular techniques or by DNA typing with allele‐specific oligonucleotide probes. These specificities were recently reported to have significant importance in antigen presentation. To avoid a time‐consuming hybridization procedure, we have developed a simple typing system using PCR and subsequent digestion by allele‐specific restriction endonucleases. A system was established with locus‐specific amplification of HLA‐DRB3 and digestion by the enzymes KpnI, Seal and HinfI which recognize unique restriction sites within the amplified region. This allowed HLA‐DRB3 typing on agarose gel by determining whether the amplification product has been digested or not. This typing system was compared to conventional oligotyping by analyzing 145 RFLP‐typed individuals for their DRw52 specificity using both methods. Agarose typing correlated well with oligotyping and was shown to be more simple and practical even in

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01873.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Abstract:In spite of a number of investigations, the concept of human leukocyte antigen (HLA) sharing in recurrent spontaneous abortion (RSA) couples remains controversial. We introduced the basal antigen sharing rate (BSR) by the original mathematical approach using the gene frequency of all HLA specificities derived from our regional control population and applied this parameter for the comparison with RSA couples. RSA couples were classified into three subgroups; primary (3 or more consecutive abortions), secondary (3 or more consecutive abortions after 1 live birth), and potential (2 consecutive abortions) aborters. No significant differences between the HLA class I sharing rates (one or more antigens shared on a single locus) of the all RSA subgroups and the calculated BSRs were observed. In the HLA‐DR and DQ loci, on the other hand, the antigen sharing rates of primary aborters were significantly higher than BSRs (p<0.01/DR, p<0.05/DQ). While potential aborters showed a result similar to that of the primary aborters, no significant antigen sharing of HLA class II was observed in secondary aborters. Our data suggest that BSR is a useful parameter for detection of significant HLA sharing in regional populations and the consecutive abortions that occurred primarily are certainly relevant to HLA class II sharin

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01874.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Abstract:Eighteen unrelated Chinese patients with insulin‐dependent diabetes mellitus (IDDM) were analyzed for HLA Class II genes using a variety of molecular biological techniques including restriction fragment length polymorphism (RFLP), polymerase chain reaction with allele‐specific oligonucleotides (PCR‐ASO) and direct DNA sequencing. The high frequency of DR3/DR4 heterozygotes found in the Chinese with IDDM strengthens the importance of this combination of haplotypes in IDDM susceptibility since it is present in two genetically distant populations ‐Chinese and Caucasians. The frequency of DRw9, a rare allele in the Caucasian population, is much higher in the Chinese. Moreover, the DQβ chain linkage of DRw9 was different in IDDM patients compared with control subjects. In contrast with previous results, codon 57 of the DQβ chain was aspartic acid in DRw9 Chinese IDDM patients. Furthermore, one particular DRw9–DQw9 haplotype may be associated with IDDM susceptibility in the Chinese

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01875.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

摘要:

Abstract:The human recombinant HLA‐DRB1 gene probe was used for histocompatibility typing of two families of beagles for the DLA‐D equivalent by using restriction fragment length polymorphisms (RFLP). This method was able to determine the segregation of these genes from the parental animals to the individual Fl offspring. Mixed lymphocyte culture (MLC) reactivity as well as serological typing for class I histocompatibility antigens were also performed for comparison. It was found that there was a high correlation between these three methods. We therefore conclude that RFLP typing is an effective procedure for predicting MLC reactivity in dogs and propose that it is a suitable genotyping method for assignment of class II antigen compatibility for donor‐recipient pairs in conjunction with organ transplant st

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01876.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01877.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01878.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01879.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1991.tb01880.x

出版商:Blackwell Publishing Ltd    

年代:1991

数据来源: WILEY