基于PDANPs和T4 PNK、λexo生物传感技术建立

Establishment of Biosensors Based on PDANPs and T4 PNK,λ exo

为找到一种快速灵敏检测miRNA-199的方法,分析纳米材料PDANPs表征,构建基于聚多巴胺纳米粒子(PDANPs)和T4多聚核苷酸激酶(T4 PNK)、λ核酸外切酶(λexo)的新型纳米生物传感技术检测miRNA-199平台,对影响生物传感技术主要因素进行优化,分析其线性范围和检测限;与茎环引物设计荧光定量PCR方法进行比对。结果显示:合成PDANPs直径300.0±31.7 nm,表面含有丰富的π电子;反应体系荧光强度与miRNA-199浓度的对数成线性关系,F=65.183 lg C+157.3、R 2=0.973;线性范围为10 pmol/mL^100 nmol/mL;检出限值为4.62 pmol/mL;与荧光定量PCR法比对结果偏倚为-0.22105 pmol/mL。结果表明,基于PDANPs和T4 PNK、λexo新型纳米生物传感技术可用于miRNA-199检测。

In order to find a fast and sensitive method to detect mirna-199,the features of PDANPs surface were analyzed.A new nano biosensor platform based on PDANPs,T4 PNK andλexo were constructed to detect mirna-199.The main factors affecting the biosensor technology were optimized,and the linear range and detection limit were analyzed.The platform was compared with the fluorescence quantitative PCR method designed by stem ring primers.The results showed that the diameter of synthesized PDANPs was 300.0±31.7 nm,and there were abundantπelectrons on the surface;The fluorescence intensity of the reaction system was linear with the logarithm of mirna-199 concentration,F=65.183lg C+157.3,R 2=0.973;The linear range was 10 pmol/mL^100 nmol/mL;The detection limit was 4.62 pmol/mL;The bias of the results was-0.22105 pmol/mL.The new nano biosensors based on PDANPs,T4 PNK andλexo can be used to detect mirna-199.