Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center, Kurume, Fukuoka, Japan.
Kahotechno Co., Ltd., Iiduka, Fukuoka, Japan.
PLoS One. 2023 Sep 21;18(9):e0286229. doi: 10.1371/journal.pone.0286229. eCollection 2023.
For early detection of canine urothelial and prostatic carcinoma, we intend to develop and commercialize a simple and rapid detection method for the BRAF V595E mutation, a known mutation in this cancer. Detection of the single-nucleotide substitution in cancer cells contained in urine sediments is effective for early cancer diagnosis. However, urine sediment also contains many normal cells, and when there is a small relative composition of cancer cells, the mutation is difficult to detect by conventional methods other than next-generation sequencing. Our new detection method enables reliable discrimination with the same labor and cost as the PCR method. We compared the results of our new method with the results of the conventional Sanger method for 38 canine urine sediment samples, and the results of 34 samples were consistent between both methods. The remaining four results were all determined to be negative by the Sanger method and positive by our new method. For these four samples, the ratio of the mutated gene to the wild-type gene was estimated using a third-generation sequencer, and the ratio of the mutated gene was 0.1%-1.4%. We postulate that the Sanger method gave a negative result because of the low abundance of the mutated gene in these samples, proving the high sensitivity of our new method.
为了早期检测犬类的尿路上皮癌和前列腺癌，我们旨在开发和商业化一种简单、快速的 BRAF V595E 突变检测方法，这是此类癌症中的一种已知突变。检测尿液沉淀物中癌细胞中的单核苷酸取代对于早期癌症诊断是有效的。然而，尿液沉淀物中也含有许多正常细胞，当癌细胞的相对组成较少时，除了下一代测序之外，传统方法很难检测到突变。我们的新检测方法可以与 PCR 方法一样可靠地进行区分，且具有相同的劳动和成本。我们将我们的新方法的结果与 38 个犬尿液沉淀物样本的常规 Sanger 方法的结果进行了比较，两种方法的结果在 34 个样本中是一致的。其余四个结果在 Sanger 方法中均为阴性，而在我们的新方法中为阳性。对于这四个样本，使用第三代测序仪估计了突变基因与野生型基因的比值，且突变基因的比值为 0.1%-1.4%。我们推测 Sanger 方法给出了阴性结果，是因为这些样本中突变基因的丰度较低，证明了我们的新方法具有较高的灵敏度。
