Soejima Mikiko, Koda Yoshiro
Department of Forensic Medicine, Kurume University School of Medicine, Kurume, Japan.
Hum Mutat. 2025 Jul 23;2025:3468799. doi: 10.1155/humu/3468799. eCollection 2025.
Three nonsynonymous single nucleotide variations (SNVs), rs1042602 in TYR (p.S192Y), rs1426654 in SLC24A5 (p.A111T), and rs16891982 in SLC45A2 (p.L374F), were associated with human skin pigmentation variation and may have recently undergone positive natural selection. Furthermore, these three SNVs have been reported to correlate with the risk and prognosis of melanoma. To simultaneously determine these three SNVs, a triplex fluorescent probe-based melting curve assay (FMCA) was developed. The method was validated by analyzing genomic DNA from subjects with known genotypes. For rs16891982, triplex FMCA did not allow good separation of genotypes with the initial polymerase enzyme mix used, but by changing the enzyme mix used, the three genotypes could be clearly distinguished. Using this method, we definitively genotyped these three SNVs in 93 European, 58 Tamil, 54 Sinhalese, and 52 Bangladeshi subjects. This method allows genotyping of rs1042602, rs1426654, and rs16891982 in a relatively large number of samples to perform association studies on skin pigmentation variation or melanoma risk.
三个非同义单核苷酸变异（SNV），即TYR基因中的rs1042602（p.S192Y）、SLC24A5基因中的rs1426654（p.A111T）以及SLC45A2基因中的rs16891982（p.L374F），与人类皮肤色素沉着变异相关，且可能最近经历了正向自然选择。此外，据报道这三个SNV与黑色素瘤的风险和预后相关。为了同时检测这三个SNV，开发了一种基于三重荧光探针的熔解曲线分析方法（FMCA）。通过分析已知基因型受试者的基因组DNA对该方法进行了验证。对于rs16891982，最初使用的聚合酶混合物进行三重FMCA时无法很好地分离基因型，但通过改变使用的酶混合物，可以清晰区分三种基因型。使用该方法，我们对93名欧洲人、58名泰米尔人、54名僧伽罗人和52名孟加拉国受试者的这三个SNV进行了明确的基因分型。该方法能够对相对大量样本中的rs1042602、rs1426654和rs16891982进行基因分型，以开展关于皮肤色素沉着变异或黑色素瘤风险的关联研究。
