CAJ | 학술논문

利用纳米颗粒对目标DNA的富集、分离作用以及阳离子荧光共轭聚合物良好的荧光特性,建立了一种特异性检测DNA的新方法.首先将标记有猝灭基团的DNA捕获探针修饰到纳米颗粒上,捕获互补的DNA分子;然后加入S1核酸酶,除去未捕获到互补DNA的捕获探针;最后用Dnase Ⅰ将颗粒上的双链切断,使猝灭基团从纳米颗粒上解离下来,与阳离子荧光共轭聚合物结合并猝灭其荧光.结果表明,目标核酸的浓度与该聚合物的荧光猝灭程度正相关,且具有良好的特异性,线性响应范围为5.0～40 nmol/L; 检出限为3.7 nmol/L(S/N=3).
이용납미과립대목표DNA적부집、분리작용이급양리자형광공액취합물량호적형광특성,건립료일충특이성검측DNA적신방법.수선장표기유졸멸기단적DNA포획탐침수식도납미과립상,포획호보적DNA분자;연후가입S1핵산매,제거미포획도호보DNA적포획탐침;최후용Dnase Ⅰ장과립상적쌍련절단,사졸멸기단종납미과립상해리하래,여양리자형광공액취합물결합병졸멸기형광.결과표명,목표핵산적농도여해취합물적형광졸멸정도정상관,차구유량호적특이성,선성향응범위위5.0～40 nmol/L; 검출한위3.7 nmol/L(S/N=3).
A novel method for DNA detection was developed based on the excellent fluorescence properties of cationic conjugated polymer ( CCP)and the target DNA enrichment,separation function of nanoparticles. First,the quencher-labeled DNA capture probes were modified on the surface of Au nanoparticles,and complementary DNA strands were captured. Second,S1 nuclease was added,and the capture probes that had not captured the complementary DNA were removed from the nanoparticles.Finally,the complementary double-stranded DNA was cut by Dnase I,the quenchers were dissociated from nanoparticle and the fluorescence of CCP was quenched by means of combination of quenchers and CCP.The results showed that this method is specific.In the range of 5. 0 -40 nmol/L,the concentration of target DNA was proportional to the fluorescence quenching and the detection limit was 3.7 nmol/L(S/N = 3).