MULTIPLEX PCR ANALYSIS AND GENOTYPE-PHENOTYPE CORRELATIONS OF FREQUENT APC MUTATIONS
MULTIPLEX PCR ANALYSIS AND GENOTYPE-PHENOTYPE CORRELATIONS OF FREQUENT APC MUTATIONS
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DOI：
10.1002/humu.1380050208
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发表时间：
1995-01-01
期刊：
影响因子：
3.9
通讯作者：
MARIANICOSTANTINI, R
中科院分区：
文献类型：
--
作者：
CAMA, A;PALMIROTTA, R;MARIANICOSTANTINI, R
大肠腺瘤性息肉病（APC）基因的种系突变倾向于聚集在离散的区域。其中一些突变经常发生在家族性大肠腺瘤性息肉病（FAP）患者中，该疾病的遗传诊断策略应该包括简单的检测方法。我们研究了来自31个不相关的FAP家系的48名FAP患者或“高危”成员。采用琼脂糖微凝胶异双工分析（HAAM）和多重等位基因特异性PCR对无亲缘关系的患者进行分析。这种新策略容易且可靠地检测到密码子1061、1068和1309处三种常见的APC缺失，从而在9名不相关的患者中鉴定出突变等位基因。基于HAAM和扩增难解突变系统（ARMS）的靶向突变分析，可以在多重PCR和HAAM检测到突变的患者亲属中快速鉴定出另外11名生殖系缺失的受试者。使用两种独立的基于pcr的测试，采用不同的引物，减少了在DNA扩增过程中出现的伪影可能干扰诊断评估的可能性。基因型-表型相关性的分析提供了证据，表明在具有相同突变的受试者中，结肠和结肠外表现的程度存在异质性。然而，与密码子1061和1068突变的患者相比，密码子1309缺失与更严重的结肠疾病的一致关联，支持突变位点与FAP外显率之间的相关性。(C) 1995 Wiley-Liss, Inc。
Germline mutations of the adenomatous polyposis coli (APC) gene tend to cluster in discrete regions. Some of these mutations occur frequently in familial adenomatous polyposis coli (FAP) patients, and strategies for genetic diagnosis of the disease should include simple methods for their detection, We studied a total of 48 FAP affected or ''at-risk'' members from 31 unrelated FAP pedigrees. Unrelated patients were analyzed using heteroduplex analysis on agarose minigels (HAAM) and multiplex allele-specific PCR. This novel strategy readily and reliably detected the three frequently occurring APC deletions at codons 1061, 1068, and 1309, allowing identification of mutant alleles in nine unrelated patients. A targeted mutational analysis, based on HAAM and amplification refractory mutation system (ARMS), allowed the rapid identification of 11 additional subjects with germline deletions, among relatives of the patients in whom mutations had been detected by multiplex PCR and HAAM. The use of two independent PCR-based tests, employing distinct sets of primers, reduces the possibility that artifacts occurring during DNA amplification may interfere with the diagnostic evaluation, The analysis of genotype-phenotype correlations provided evidence for heterogeneity with regard to the extent of colonic and extracolonic manifestations of the disease in subjects bearing identical mutations. However, the consistent association of the deletion at codon 1309 with more severe colonic disease than that observed in patients with mutations at codons 1061 and 1068, supports a correlation between mutation site and penetrance of FAP. (C) 1995 Wiley-Liss, Inc.