PCR-SSP技术检测乳腺癌易感基因

[目的]建立简易的PCR方法检测BRCA1及其等位基因.[方法]根据BRCA1基因(EMBL/GenBank/DDBJ U14680)和在中国人乳腺癌患者中高频存在的BRCA1-2201T2430C2731T3232G3667G等位基因(AY304547)序列,设计二对特异性引物,分别针对正常BRCA1和上述等位基因,同时引入一对内对照引物,建立简易的序列特异性引物-pCR技术(PCR-SSP),并用于检测30份已知BRCA1序列的乳腺癌患者DNA样本,以及67名未知的健康女性捐血者样本.[结果]30份已知的患者DNA样本的PCR-SSP检测结果与样本的序列完全相符.67名健康女性检测结果显示6名个体(9.0%)为等位基因纯合体,32人(47.8%)携带正常BRCA1,其余29名个体(43.3%)为杂合体.[结论]PCR-SSP方法能特异性检测BRCA1基因和BRCA1-2201T2430C2731T3232G3667G等位基因,可作为检测该等位基因以及判断个体在这一突变热点位点是否发生变异的简易手段.

A PCR-SSP Method for Determination of BRCA1

To establish an easy method for the determination of BRCA1 and its allele.Based on the sequences of BRCA1 (EMBL/GenBank/DDBJ U14680) and the allele of BRCA1-2201T2430C2731T3232G3667G(AY304547),which is highly frequent in the Chinese patients with breast cancer,two pairs of primers were designed specific for the allele and BRCA1.With a pair of internal control primer,a simple sequence-specific primer PCR (PCR-SSP) method was established.And it was evaluated with sequenced samples from 30 breast cancer patients and unknown samples from 67 healthy female blood donors.The results of the PCR-SSP for 30 sequenced samples were absolutely concordant with each sample's sequence.The results of the 67 healthy female donors showed that 6 (9.0%) were homozygote of the allele, 32 (47.8%) carried normal BRCA1 gene, and the rest 29 (43.3%) were heterozygote.[Conclusions]PCR-SSP was specific for BRCA1 and the BRCA1-2201T2430C2731T3232G3667 allele, it might be a simple test for the allele and for BRCA1 mutation detection in breast cancer patient.