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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 establish rapid detection of measles virus nucleic acid by Helicase-dependent Isothermal Deoxyribonucleic Acid (DNA) Amplification (HDA) technique. Methods Specific primers were designed on the hemagglutinin gene of measles virus, and were amplified by RT-tHDA reverse transcript-therm ophilic HDA. The reaction conditions optimize. Results The detection of measles virus nucleic acid by RT-tHDA did not cross-react with rubella, mumps, enterovirus 71 and respiratory syncytial virus, and had good specificity. The detection limit was 5.012 × 10-10 mol / L (mol / L), the sensitivity and ordinary RT-PCR method no significant difference. Conclusion The detection method of measles virus RT-tHDA has the characteristics of simple, specific, sensitive and low requirement for the instrument, which provides a new method for the detection of the nucleic acid of measles virus.