ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03126.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

Abstract: We have developed a single DNA typing method which uses 144 sequence‐specific primer (SSP) reactions to simultaneously detect all known HLA‐A, B, C, DRB1, DRB3, DRB4, DRB5 and DQB1 specificities in an allele specific or group specific manner using the same method, reagents, PCR parameters and protocols for all loci. The results from this integrated class I&II method can be visualized on a single photographic or electronic image and hence is described as “Phototyping”. Phototyping has an overall resolution greater than or equivalent to good serology and results can be obtained in under 3 hours making the method suitable for genotyping potential cadaver donor peripheral blood without serological backup. This in turn produces the potential for reducing cold ischaemia times in renal transplantation as well as the application of prospective matching to cardiac and liver transplantation. The method has capacity to detect new alleles, for example, novel amplification patterns suggestive of 4 new HLA‐B alleles have been detected. The Phototyping set has been used as the sole method of HLA typing for over 1010 individuals. Phototyping is not problem‐free; deviations from the standard protocol, poor quality DNA and unsuitable PCR machines can result in individual PCR failures or in incorrect assignment of antigens. Approximately 5% of genotypes were repeated (either partially or fully) because of incomplete or equivo

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03127.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

Abstract: Current typing technology for class I HLA antigens uses serological and/or isoelectric focusing gel electrophoresis. DNA typing for the HLA class I antigens can accurately identify the class I genotype of individuals and cell lines. Here, we report correlation of DNA typing results with serological and IEF results for the B17 group. The B17 antigens are relatively common, being carried by almost 9% of Caucasians and 28% of blacks. In this study, five 10th International Histocompatibility Workshop cell lines carrying B17 and 106 individuals in 61 families carrying B17 were DNA typed for B17 using B17‐allele‐specific amplification and sequence specific oligonucleotide probe hybridization pattern analysis. 38 (55.07%) out of 69 unrelated haplotypes had B*5701, 23 (33.33%) had B*5801, 6 (8.70%) had B*5702, and 2 (2.90%) had B*5802. DNA typing results correlated well with serological and isoelectric focusing results. In general, there was high degree of agreement between all three methods, although heterozygosity for B17 poses a particular problem for serological and IEF methodology. Both B*5701 and B*5801 have the same electrophoretic mobility on IEF gel, corresponding to B17.2, B*5702 corresponds to B17.1, while B*5802 corresponds to B1

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03128.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

Abstract:Neonatal alloimmune thrombocytopenia (NATP) and post‐transfusion purpura (PTP) are acquired bleeding disorders caused by alloimmune thrombocytopenia. In most cases, the thrombocytopenia is due to an alloantibody directed against the platelet glycoprotein IIb‐IIIa (GPIIb‐IIIa) complex. During the course of routine studies on the role of GPIIb‐IIIa in inherited and acquired bleeding and thrombotic disorders, we unexpectedly identified an individual whose platelets reacted by non‐reduced Western blot analysis with anti‐GPIIIa polyclonal antisera, but did not react with a commercially available monoclonal antibody (SZ21) specific for GPIIIa. We screened all 14 GPIIIa exons for possible nucleotide changes which might alter amino acids and found variations in only exons 3 and 10. Nucleotide sequencing revealed that only the exon 3 alteration changed the predicted amino acid sequence. This variation was caused by homozygosity for the uncommon PlA2allele of the GPIIIa gene. Platelets from two additional unrelated normal individuals known to be homozygous for PlA2also lacked reactivity with SZ21 by Western blot. Using flow cytometry with intact platelets, we observed a markedly reduced binding of SZ21 to platelets with the PlA2genotype. Scatchard analyses indicated that SZ21 bound to P1A1/A1platelets with a Kd of 8.26times 10‐10M, and to P1A2/A2platelets with a Kd of 5.58times10‐9M. Thus, we have characterized a readily available monoclonal antibody able to distinguish between the two P1Aalleles of the GPIIIa gene. Because incompatibility for this platelet polymorphism is the most common cause of neonatal alloimmune thrombocytopenia and posttransfusion purpura, and because platelet immunophenotyping reagents lack specificity and are not easily available, this monoclonal antibody could facilitate the management of patients with

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03129.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

Abstract:It was the purpose of this study to better define the frequency of HLA‐B27 subtypes and HLA class II alleles among indigenous populations from the eastern tip of the Chukotka Peninsula of Siberia, Russia, which have higher frequencies of HLA‐B27 (40%) and spondyloarthropathies (2%) than Caucasian populations and test the hypothesis that these populations are more closely related to Orientals. Siberian Eskimos and Chukchi residing in four coastal villages on the Chukotka Peninsula inhabited by Siberian Eskimos and Chukchi people were examined using oligotyping of the polymerase‐chain reaction‐amplified second and third exons of the HLA‐B27 gene. HLA‐class II alleles (DRB1, DQA1 and DQB1) were similarly determined. Of 88 HLA‐B27 positive individuals from these villages, all had HLA‐B*2705, including the four patients with Reiter's syndrome and the five ankylosing spondylitis, except one Eskimo control who had HLA‐B*2702. None had HLA‐B*2704, a frequent subtype in Orientals. HLA‐class II typing in 70 Siberian Eskimos and 71 Siberian coastal Chukchi revealed HLA‐DRB1*0401, DRB1*0802, *0901 and *1402 to account for nearly all the DRB1 alleles found in this population, similar to what has been described in Eskimos in Alaska, but different from Chinese or native Americans in the U.S. The overwhelming majority of the individuals examined had HLA‐DQB1*0301, similar to what has been observed in Native Americans. The Siberian Eskimos differed from the coastal Chukchi only in the occurrence of HLA‐DRB1*0701, DQA1*0201, DQB1*0201 alleles, which occurred only in the former group. These data suggest that the Chukotka population is genetically more closely related to Caucasians and native Americans and less to

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03130.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

Abstract:Some combinations of HLA‐A,‐B and ‐DR antigens occur more frequently than would be expected from their gene frequencies in the population. This phenomenon, referred to as Linkage Disequilibrium (LD) has been the origin of many speculations. One hypothesis to explain LD is that some haplotypes are protected from recombination. A second hypothesis is that these HLA antigens preferentially recombine after cross‐over, to create an LD haplotype. We tested these 2 hypotheses: from a pool of over 10,000 families typed in our department, we analyzed 126 families in which HLA‐A:B or B:DR recombinant offspring was documented. To overcome a possible bias in our material, we used the non‐recombined haplotypes from the same 126 families as a control group. Our results show that the number of cross‐overs through LD haplotypes is not significantly lower then would be expected if recombination occurred randomly. Also the number of LD haplotypes created upon recombination was not significan

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03131.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03132.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03133.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03134.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

ISSN:0001-2815    

DOI:10.1111/j.1399-0039.1995.tb03135.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY