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 。
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.
出处
《中国疫苗和免疫》
CAS
2012年第6期493-495,540,共4页
Chinese Journal of Vaccines and Immunization
基金
浙江省重大科学技术专项重点社会发展项目"疫苗时代麻疹流行因素与预防控制技术研究"课题(编号:2009C03001-4)
浙江省公共卫生检验检测重点学科群(编号:XKQ-009-003)
浙江省自然科学基金"麻疹流行株血凝素基因变异与病毒抗原性改变的关系研究"(编号:Y2091178)
关键词
麻疹病毒
逆转录-依赖耐热解旋酶恒温扩增技术
检测
Measles virus
Reverse transcription-thermophilic helicase-dependent isothermal deoxyribonucleic acid amplification
Detection