【摘要】 目的建立依赖解旋酶恒温扩增[Helicase-dependent Isothermal Deoxyribonucleic Acid(DNA)Amplification,HDA]技术快速检测麻疹病毒核酸。方法 在麻疹病毒的血凝素基因上设计特异性引物,采用逆转录-依赖耐热解旋酶恒温扩增(Reverse Transcription-therm ophilic HDA,RT-tHDA)技术进行恒温扩增,并对反应条件进行优化。结果采用RT-tHDA技术检测麻疹病毒核酸,对风疹病毒、流行性腮腺炎病毒、肠道病毒71型、呼吸道合胞病毒均无交叉反应,具有良好的特异性;且最低检测限可达5.012×10-10摩尔/升(mol/L),敏感度与普通RT-聚合酶链反应方法 无明显差别。结论麻疹病毒RT-tHDA检测方法 具有简便、特异、敏感以及对仪器要求低的特点,为麻疹病毒的核酸检测,提供了一种新方法 。更多还原

【Abstract】 Objective To develope a helicase dependent isothermal deoxyribonucleic acid (DNA) amplification assay for quick detection of measles virus.Method A rapid one step reverse transcription thermophilic helicase dependent isothermal DNA amplification (RT tHDA) assay was developed using primers pairs designed within hemagglutinin gene of measles virus and the reaction systems were optimized.Results There were no crossing reactions with mumps virus,rubella virus,human enterovirus type 71 and espiratory syncytial virus,which indicated the good specificity of the RT tHDA assay.Sensitivity of the RT tHDA,5.012×10-10mol/L,has no significant difference compared with conventional RT Polymerase Chain Reaction assay.Conclusions RT tHDA assay is simple,specific,sensitive and no extra requirement for instrument,which provide a new approach for quick detection of measles virus.更多还原