CAJ | 학술논문

基于急性早幼粒细胞白血病(APL)中PML/RARα融合基因的碱基序列,设计了新型的锁核酸(LNA)修饰寡核苷酸作为捕获探针和信号探针,研究出一种基于"三明治"传感模式的电化学生物传感器对PML/RARα融合相关基因进行检测.靶序列分别与捕获探针和信号探针杂交后形成"三明治"结构.将修饰电极置于含有底物3,3′,5,5′-四甲基联苯胺(TMB)和过氧化氢的测定溶液中,用计时电流法检测靶序列.结果表明,该传感器可定量识别和检测溶液中人工合成的短链APL PML/RARα融合基因片段.经过条件优化,杂交前后电流值与靶标链浓度在1.0×10~(-12) ～2.5×10~(-11) mol/L范围内呈良好的线性关系,检出限为8.5×10~(-13) mol/L.该方法简单、特异性好,有望用于实际样品的检测.
기우급성조유립세포백혈병(APL)중PML/RARα융합기인적감기서렬,설계료신형적쇄핵산(LNA)수식과핵감산작위포획탐침화신호탐침,연구출일충기우"삼명치"전감모식적전화학생물전감기대PML/RARα융합상관기인진행검측.파서렬분별여포획탐침화신호탐침잡교후형성"삼명치"결구.장수식전겁치우함유저물3,3′,5,5′-사갑기련분알(TMB)화과양화경적측정용액중,용계시전류법검측파서렬.결과표명,해전감기가정량식별화검측용액중인공합성적단련APL PML/RARα융합기인편단.경과조건우화,잡교전후전류치여파표련농도재1.0×10~(-12) ～2.5×10~(-11) mol/L범위내정량호적선성관계,검출한위8.5×10~(-13) mol/L.해방법간단、특이성호,유망용우실제양품적검측.
A new kind of sandwich-type DNA electrochemical biosensor is designed to detect PML/RARα fusion gene in acute promyelocytic leukemia.This biosensor is based on a "sandwich" detection strategy,in which involves a pair of locked nucleic acids(LNA) modified oligonucleotides as probes,including capture probe DNAs and reporter probe DNAs,both of which contain complementary sequences to one end of the target DNA.An enzymatically amplified electrochemical current signal was observed when the biosensor was used for detection of target DNA in sample containing 3,3′,5,5′-tetramethylbenzidine(TMB) and H_2O_2 substrate.The results indicate that,under the optimal conditions,the relationship between amperometric current and the concentration of synthetic target DNA complementary strand is linear in the range of 1.0×10~(-12)-2.5×10~~(-11) mol/L with a detection limit of 8.5×10~(-13) mol/L.The method show good simplicity and specificity,and is expected to be used in real sample.