A primerless amplification suitable for enrichment of particular genotype cfDNA which is a one-dimensional material has been developed. This primerless amplification coordinated by two thermostable enzymes of endonuclease and proofreading polymerase, functions as a genotype switch in analyzing cfDNA. The endonuclease digests the wild-typed fragments into mega-primer and discriminately destroys the wild-type DNA alleles. The DNA polymerase proofreads the megaprimer and then extends the mega-primer using the mutant DNA as the template. The prototypes of this technology were applied to two hotspot mutations of APC and EGFR with confirmed by DNA sequencing analysis. Genotype switch was then employed to clinical cfDNA assay targeting PIK3CA. Data from the clinical application suggest its potential in early cancer diagnosis.

中文翻译：

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循环肿瘤DNA分析的无底漆扩增。

已经开发了适合于富集作为一维材料的特定基因型cfDNA的无引物扩增。这种无引物的扩增由核酸内切酶和校对聚合酶的两种热稳定酶协调，在分析cfDNA时起基因型转换的作用。核酸内切酶将野生型片段消化成大型引物，并区别破坏野生​​型DNA等位基因。DNA聚合酶对超级引物进行校对，然后使用突变的DNA作为模板扩展超级引物。经DNA测序分析证实，该技术的原型被应用于APC和EGFR的两个热点突变。然后将基因型转换用于靶向PIK3CA的临床cfDNA检测。来自临床应用的数据表明其在早期癌症诊断中的潜力。