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A novel biomarker for acute kidney injury using TaqMan-based unmethylated DNA-specific polymerase chain reaction

机译：基于TaqMan的未甲基化DNA特异性聚合酶链反应的急性肾脏损伤的新型生物标记

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摘要

There has been increasing interest in the use of circulating DNA as biomarkers for various tissue injuries, cancers, and fetal conditions. DNA methylation is a well-characterized mechanism underlying the epigenetic regulation of gene expression, and many diagnostic tests based on DNA methylation patterns have been developed. We developed a novel TaqMan-based assay for the detection of acute kidney injury using a hypomethylated promoter region of Slc22a12, a urate transporter specifically expressed in proximal tubular cells. Bisulfite sequencing analysis confirmed that the CpG islands in the promoter region of mouse Slc22a12 were preferentially hypomethylated in the kidney cortex. TaqMan minor groove binder(MGB) probes reliably discriminated the DNA fragments corresponding to the unmethylated and methylated promoter regions of Slc22a12. Plasma levels of unmethylated DNA corresponding to the Slc22a12 promoter region were undetectable at baseline and were significantly elevated after acute kidney cortex necrosis. This study showed the usefulness of the TaqMan system in discriminating methylated and unmethylated DNA fragments, and the similar strategy can be applied for establishing biomarkers for various cellular injuries or pathological conditions.
机译：使用循环DNA作为各种组织损伤，癌症和胎儿疾病的生物标志物的兴趣日益浓厚。 DNA甲基化是基因表达的表观遗传学调控基础上的一个公认的机制，并且已经开发了许多基于DNA甲基化模式的诊断测试。我们开发了一种新颖的基于TaqMan的检测方法，用于检测Slc22a12的低甲基化启动子区域（一种在近端肾小管细胞中特异性表达的尿酸盐转运蛋白）的低甲基化启动子区域。亚硫酸氢盐测序分析证实，小鼠Slc22a12启动子区域中的CpG岛优先在肾皮质中被低甲基化。 TaqMan小沟结合剂（MGB）探针可可靠地区分与Slc22a12的未甲基化和甲基化启动子区域相对应的DNA片段。对应于Slc22a12启动子区域的未甲基化DNA的血浆水平在基线时未检测到，并在急性肾皮质坏死后显着升高。这项研究表明TaqMan系统在区分甲基化和未甲基化的DNA片段中的有用性，并且类似的策略可以用于建立针对各种细胞损伤或病理状况的生物标记。

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