Duplex dual‐labeled fluorescence probe‐based melting curve and endpoint genotyping assays for genotyping of rs2000999 and haptoglobin gene deletion
Duplex dual‐labeled fluorescence probe‐based melting curve and endpoint genotyping assays for genotyping of rs2000999 and haptoglobin gene deletion
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DOI：
10.1002/elps.202200202
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发表时间：
2022-10
期刊：
影响因子：
2.9
通讯作者：
M. Soejima;Y. Koda
中科院分区：
文献类型：
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作者：
M. Soejima;Y. Koda
Haptoglobin （Hp）是一种结合血红蛋白的血清糖蛋白。Hp基因（Hp）或Hp相关基因（HPR）的一些变异，包括HRP内含子2的单核苷酸多态性（rs2000999）和Hp基因（HPdel）的完全缺失（HPdel是Hp的一种罕见变异），已被报道与血清胆固醇浓度和血清Hp浓度相关。在本研究中，我们建立了一种基于双标记荧光探针的方法，通过熔融曲线基因分型同时检测rs2000999 G > a多态性，通过终点基因分型同时检测HPdel的合子性。然后使用94名日本受试者的基因组DNA验证了该方法，这些受试者的rs2000999和HPdel合子性基因型已经确定。所得结果与前人的结果完全吻合。因此，目前的方法使我们能够在相对大规模的受试者群体中估计这两种多态性，特别是在分布HPdel的亚洲人群中。
Haptoglobin (Hp) is a hemoglobin‐binding serum glycoprotein. Some variations in the Hp gene (HP) or Hp‐related gene (HPR), including a single‐nucleotide polymorphism in intron 2 of HRP, rs2000999, and a complete deletion of the HP gene (HPdel), one of the rare variants of HP, have been reported to correlate with the serum cholesterol concentration as well as the serum Hp concentration. In this study, we developed a duplex dual‐labeled fluorescence probe‐based method to simultaneously determine the rs2000999 G > A polymorphism by melting curve genotyping and the zygosity of HPdel by endpoint genotyping. This method was then validated by using the genomic DNA from 94 Japanese subjects for whom genotypes of rs2000999 and HPdel zygosity had already been determined. The results obtained with this method were in perfect agreement with the previous ones. Thus, the present method enables us to estimate these two polymorphisms in relatively large‐scale groups of subjects, especially in Asian populations where the HPdel is distributed.