化学发光核酸传感器的研制

Chemiluminesencent Biosensor For Nucleic Acid

用N-乙酰半胱氨酸金表面自组装技术，以及用1-乙基3-（3-二甲氨基）碳二亚胺盐酸盐（EDC）、N-羟基硫代琥珀酰亚胺（NHS）偶联剂把亲合素固定于金表面，再将生物素标记深外固定在亲合素上，来制备核酸传感器探头。然后将探头与目标DNA和第二生物素标记探针进行夹心式杂交，再利用生物素-亲合素的作用联人亲合素标记的碱性磷酸酯酶，用酶催化底物AMPPD发光来达到测定DNA的目的。测定了乙型肝炎病毒（HBV）DNA（566bp）的片段，实验测得HBV-DNA灵敏度为3×10^(-14)mol/L。

A chemiluminescent biosensor for nucleic acid was descrital. Cold surface was modfied with N-Acetylcysteine to produce a self-assembled monolayer. A single layer film of biotinylated olignucleotide probes was constructed by binding to a precursor layer of streptavidin which had been immobilized on modified gold surface covalently using carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide(NHS). A sandwich solution hybridization was executed with immobilized deoxyribo nucleic acid (DNA), target DNA and second biotinylated DNA probes. Then extravidin alkaline phosphatase which can dephosphorylate substrates AMPPD to produce stable intermediates that produce a measurable yield of light output was bound with second hybrided biotinylated DNA probes. And hepatitis B virus (HBV) DNA fragment was meased. The sensitivity of HBV DNA was obtained as 3 ×10^(-14) mol/L.