目的：建立一种简便、快速并且准确地检测出 BRCA1基因突变的检测技术，以了解 BRCA1的遗传突变与乳腺癌发病的关系。方法采用高分辨率熔解曲线（HRM）技术，对确诊的30例乳腺癌患者以及10例正常女性的全血样本，进行 BRCA1基因的三个重要的遗传学突变（185 delAG、4956 A > G 以及1100 delAT）的 HRM 检测。并且同时采用传统的测序法，对其进行进一步的验证。结果30例确证乳腺癌患者中185 delAG 纯合突变2例，杂合突变1例；4956 A > G 纯合突变3例，杂合突变5例，1100 delAT 纯合突变2例，杂合突变3例。将所有40例样本进行 BRCA1基因直接测序验证，所有经过 HRM 检测纯合突变确实产生了碱基的缺失，杂合突变的样本中突变或缺失导致的移码突变所形成的双峰。测序获得的比对结果与 HRM 结果完全一致。结论 HRM 可以快速有效地检测出已知的 BRCA1基因突变，与测序结果完全一致，可应用于 BRCA1的临床快速检测中。%Objective Genetic mutations in the BRCA1 and incidence of breast cancer is often closely related. We need conventional de-tection techniques to getting simple,fast and accurate detection of mutations in the BRCA1 gene. Methods We use a high - resolution melting curve(HRM)technology in whole blood samples of 30 breast cancer patients and 10 healthy patients with definite diagnosis. Three important ge-netic mutations in the BRCA1 gene(185 delAG,4956 A > G and 1 100delAT)were tested by HRM testing. And at the same time using the tradi-tional sequencing method further verification was carried out. Results 30 cases of confirmed breast cancer patients homozygous mutation in 185 delAG two cases,one case of heterozygous mutation;4956 A > G homozygous mutation in 3 cases,5 cases of heterozygous mutations,1 100 delAT homozygous mutation in 2 cases,heterozygous mutation in 3 cases. All samples were 40 cases of direct sequencing of the BRCA1 gene,all through HRM homozygous mutation does have a base deletion,heterozygous mutations in the sample cause mutation or deletion bimodal frameshift mutation formation. Sequencing results obtained compared with the HRM were completely consistent. Conclusion It showed that HRM can quickly and ef-ficiently detect known BRCA1 gene mutation,fully consistent with the sequencing results. HRM technology can be applied to rapid clinical detec-tion of BRCA1.
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