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| 基于微流控核酸等温扩增的
登革病毒现场快速检测技术研究 |
| Rapid and spot detection of dengue virus using microfluidic nucleic acid isothermal amplification |
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| DOI：10.3969/j.issn.1007-8134.2022.06.003 |
| 中文关键词: 环介导等温扩增技术 微流控 登革病毒 快速检测 |
| 英文关键词: LAMP microfluidics dengue virus rapid detection |
| 基金项目:军队青年培育孵化项目（16QNP149） |
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| 中文摘要: |
| [摘要]?目的?结合环介导等温扩增技术（loop-mediated isothermal amplification, LAMP）和微流控芯片技术，建立一种适合现场快速检测登革病毒的方法。方法?利用RT-LAMP，针对登革病毒基因组中3’非编码区中特异性序列进行扩增，建立基于微流控芯片技术的LAMP检测方法，优化检测体系，并对该方法的灵敏性和特异性进行评估。结果?基于LAMP 和微流控芯片技术的登革病毒检测方法，通过对病毒模板进行扩增，发现与其他病毒无交叉反应，特异性良好；同时，结果显示该方法对登革病毒检测灵敏性可达61.2 pg/μl，与实时荧光定量PCR仪所达到的检测灵敏性一致。结论?基于LAMP和微流控芯片技术的登革病毒检测方法具有操作简单、快速、对设备要求低等优势,并且灵敏性、特异性均较好，是一种便于开展现场快速检测的方法。 |
| 英文摘要: |
| [Abstract] Objective?Establish and evaluate a technology of rapid and spot molecular detection of dengue virus using loop-mediated isothermal amplification detection and microfluidic chip. Methods?Amplifyingication of the 3’ non-coding region (NCR) of the dengue virus by integrating RT-LAM reserve transcription loop-mediated isothermal amplification (RT-LAMP), with microfluidic chip technology and optimizing the reaction condition, a detection method based on RT-LAMP and microfluidic chip technology was established, which was evaluated sensitivity and specialty. Results?Based RT-LAMP and microfluidic chip technology, we have established a method to detect dengue virus, which had a greatspecialty and had no cross amplification with other RNA virus. Meanwhile, the results suggested the sensitivity of the RT-LAMP assay in microfluidic chip was 61.2 pg/μl, which was same as the sensitivity of RT-PCR. Conclusions?The established method which was based on RT-LAMP and microfluidic chip technology had simple operations and low equipment requirements as well as great sensitivity andspecialty, it could be used for on-site detection. |
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