PNA与引物的连接方法以及探讨潜在的价值  

Linking methods of PNA to primers and exploring their potential value

作　　者：农琛 刘德文 赖凤明 王太重[2] 唐玉莲[2] Nong Chen;Liu Dewen;Lai Fengming;Wang Taizhong;Tang Yulian(Graduate School,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China;School of Laboratory Science,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院研究生学院,广西百色533000 [2]右江民族医学院检验学院,广西百色533000

出　　处：《右江民族医学院学报》2023年第2期263-270,共8页Journal of Youjiang Medical University for Nationalities

基　　金：广西自然科学基金项目(2019XNSFBA245101);广西高校中青年教师科研基础能力提升项目(2019KY0575)。

摘　　要：目的探讨一种基于点击化学反应将PNA和引物进行连接的新方法,挖掘PNA-引物具有选择性扩增cffDNA的潜力。方法采用点击化学将PNA与引物进行连接,通过变性聚丙烯酰胺凝胶电泳、高效液相色谱仪以及质谱分析对PNA-引物进行表征。最后,以DNA短片段和长片段分别模拟cffDNA和母源DNA,将包含相同摩尔质量的长短片段混合溶液通过聚合酶链式反应确定连接后的产物性能。结果PNA与引物成功连接,且连接物同时具备二者的特性。PNA-引物特异性富集DNA短片段的效率高达83.20%。结论本研究构建的新方法有望从高母源性DNA背景下,选择性地富集cffDNA,该方法同时具有通量高、成本低的特点,为cffDNA的分离及纯化提供新的思路。Objective To explore a new method of linking PNA to primers based on click chemical reaction and to explore the potential of PNA-primers for selective amplification of cffDNA.Methods Click chemistry was used to link the PNA to the primers,and the PNA-primers were characterized by denaturing polyacrylamide gel electrophoresis,high performance liquid chromatography,and mass spectrometry analysis.Finally,short and long fragments of DNA were used to simulate cffDNA and parental DNA,respectively.A solution containing a mixture of short and long fragments of the same molar mass was subjected to a polymerase chain reaction to determine the properties of the ligated product.Results PNA was successfully linked to the primer,and the linker had the properties of both.The PNA-primer specifically enriched short DNA fragments with an efficiency of 83.20%.Conclusion The new method constructed in this study is expected to selectively enrich cffDNA from a high maternal DNA background,which is also characterized by high throughput and low cost,and provides new ideas for the isolation and purification of cffDNA.

关 键 词：非损伤性产前诊断 肽核酸 胎儿游离DNA 选择性富集 点击化学 

分 类 号：R342.3[医药卫生—基础医学]