Center of Molecular Diagnostics and Personalized Medicine, Department of Basic Sciences, Division of Health Sciences, University of Monterrey, San Pedro Garza Garcia, Nuevo Leon 66238, Mexico.
Breast Cancer Center, Monterrey Institute of Technology and Higher Education, Monterrey, Nuevo Leon 64710, Mexico.
Mol Med Rep. 2018 Aug;18(2):1531-1537. doi: 10.3892/mmr.2018.9141. Epub 2018 Jun 6.
In 2015, according to the National Institute of Statistics and Geography (INEGI), malignant breast tumors were the first cause of cancer fatality in women (6,273 fatalities) in Mexico, whereas 2,793 fatalities in women were due to ovarian cancer. A total of 5‑10% of breast cancer and 10‑15% of ovarian cancer cases are caused by a hereditary breast‑ovarian cancer syndrome, with mutations predominantly identified in the BRCA1 and BRCA2 genes. Recently, the Mexican founder mutation BRCA1 ex9‑12del was identified (deletion of exons 9‑12 with recombination between introns 8‑12). This is the most frequently reported mutation in hereditary breast/ovarian cancer in Mexico. Current detection methods include end‑point polymerase chain reaction (PCR) and Multiplex Ligation‑dependent Probe Amplification (MLPA). In the present study a cheap, sensitive and fast detection method was developed based on quantitative PCR and two TaqMan® probes, one to detect the deletion (recombination region between introns 8 and 12), and the other one a region from exon 11. With this assay, 90 samples were able to be analyzed in 2 h using 2.5 ng of DNA/reaction at a cost of ~2‑3 USD. This method is capable of detecting positive samples for DNA deletion and excluding negative ones. Therefore, the method proposed may be a useful high‑throughput diagnostic option that could be useful in future association or prevalence studies that use large populations.
2015 年，据国家统计和地理研究所（INEGI）称，恶性乳腺肿瘤是导致墨西哥女性癌症死亡的首要原因（6273 例死亡），而 2793 例女性因卵巢癌死亡。总计 5%-10%的乳腺癌和 10%-15%的卵巢癌是由遗传性乳腺癌-卵巢癌综合征引起的，其突变主要发生在 BRCA1 和 BRCA2 基因中。最近，确定了墨西哥的一个创始性突变 BRCA1 ex9-12del（外显子 9-12 的缺失与内含子 8-12 之间的重组）。这是墨西哥遗传性乳腺癌/卵巢癌中最常报道的突变。目前的检测方法包括终点聚合酶链反应（PCR）和多重连接依赖性探针扩增（MLPA）。在本研究中，基于定量 PCR 和两个 TaqMan®探针开发了一种廉价、敏感和快速的检测方法，一个探针用于检测缺失（内含子 8 和 12 之间的重组区域），另一个探针用于检测外显子 11 的一个区域。使用该检测方法，在 2 小时内使用 2.5ng 的 DNA/反应，可对 90 个样本进行分析，成本约为 2-3 美元。该方法能够检测到 DNA 缺失的阳性样本，并排除阴性样本。因此，所提出的方法可能是一种有用的高通量诊断选择，可用于未来使用大人群进行的关联或流行率研究。
