USE OF POOLED DNA SAMPLES TO DETECT LINKAGE DISEQUILIBRIUM OF POLYMORPHIC RESTRICTION FRAGMENTS AND HUMAN-DISEASE - STUDIES OF THE HLA CLASS-II LOCI
USE OF POOLED DNA SAMPLES TO DETECT LINKAGE DISEQUILIBRIUM OF POLYMORPHIC RESTRICTION FRAGMENTS AND HUMAN-DISEASE - STUDIES OF THE HLA CLASS-II LOCI
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DOI：
10.1073/pnas.82.20.6970
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
ERLICH, H
中科院分区：
文献类型：
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作者：
ARNHEIM, N;STRANGE, C;ERLICH, H
一种快速的方法已经开发出来，并用于搜索限制性片段长度多态性（RFLPs）的连锁不平衡与疾病相关的基因座。通过使用基因组印迹杂交分析DQ β-链和DR β-链cDNA探针，我们研究了与胰岛素依赖型糖尿病（IDDM）易感性相关的HLA II类基因座内的DNA多态性。为了便于寻找信息丰富的RFLP，我们比较了来自IDDM患者的合并DNA样本与来自随机选择的对照个体的合并DNA样本，而不是使用传统的方法检查两组个体的DNA样本。(The附录中从理论上讨论了这种方法的适用条件。）利用这种经济、快速的方法，获得了与IDDM相关的几个特异性多态性限制性片段。然后，使用在该筛选中鉴定为信息性的限制性酶和探针分析HLA-DR分型的IDDM家族、纯合分型细胞和无关个体，以确定特异性限制性片段与HLA-DR血清学分型的关联以及在对照和IDDM人群中的频率。一些个别的多态性片段，胰岛素依赖型糖尿病人群富集与HLA-DR 3强相关，而其他人与HLA-DR 4强相关。一些片段（例如，用DR β检测的10-腺苷酸酶Taq I片段。探针）在IDDM人群中更普遍细分了血清学定义的HLA-DR类型，并可能是IDDM易感性的信息标记。
A rapid method has been developed and used to search for restriction fragment length polymorphisms (RFLPs) that are in linkage disequilibrium with disease-associated loci. By using genomic blot-hybridization analysis with DQ .beta.-chain and DR .beta.-chain cDNA probes, we examined DNA polymorphisms within the HLA class II loci associated with susceptibility to insulin-dependent mellitus (IDDM). To facilitate the search for informative RFLPs, we compared pooled DNA samples from IDDM patients with pooled DNA samples from randomly selected control individuals, instead of using the conventional approach of examining DNA samples from individuals in two groups. (The conditions under which this approach is useful are treated theoretically in the Appendix.) Several specific polymorphic restriction fragments associated with IDDM were revealed by using this economical and rapid approach. The restriction enzymes and probes identified as informative in this screening were then used to analyze HLA-DR-typed IDDM families, homozygous typing cells, and unrelated individuals to determine the association of the specific restriction fragments with HLA-DR serological type and the frequency in control and IDDM populations. Some individual polymorphic fragments for which the IDDM population was enriched correlated strongly with HLA-DR3, whereas others correlated strongly with HLA-DR4. Some fragments (e.g., a 10-kilobase Taq I fragment detected with the DR.beta. probe) that were more prevalent in the IDDM population subdivided the serologically defined HLA-DR type and may be informative markers for IDDM susceptibility.