依赖解旋酶恒温扩增技术快速检测麻疹病毒核酸

Quick Detection of Measles Virus by Helicase-dependent Isothermal Deoxyribonucleic Acid Amplification Assay

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 。

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 syneytial virus, which indicated the good specificity of the RT-tHDA assay. Sensitivity of the RT-tHDA, 5.012 × 10^-10 mol/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.