| 基于实时荧光环介导等温扩增的人乳头瘤病毒58型基因检测 |
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| 引用本文： | 黄月明,周骏,牟颖,毕瑩,张智琪,吴青青.基于实时荧光环介导等温扩增的人乳头瘤病毒58型基因检测[J].现代预防医学,2020,0(23):4326-4330. |
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| 作者姓名： | 黄月明 周骏 牟颖 毕瑩 张智琪 吴青青 |
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| 作者单位： | 1.贵州医科大学微生物与免疫学教研室，贵州 贵阳550004；2. 浙江大学工业控制技术国家重点实验室/智能系统与控制研究所/分析仪器研究中心；3.贵州医科大学附属医院临床检验中心 |
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| 摘 要： | 目的 建立基于实时荧光环介导等温扩增的人乳头瘤病58型基因检测体系，用于其快速检测和早期诊断。 方法 运用LAMP在线引物设计软件Primer Explorer V5, 针对人乳头瘤病毒58型的L1区域保守基因设计引物，在反应前加入荧光核酸染料SYBR Green Ⅰ，并对该反应体系中的dNTPs、Mg2+和内引物（FIP、BIP）浓度进行优化,优化结果通过“S”型扩增曲线判断，并对优化后的实时荧光环介导等温扩增体系进行特异性、灵敏度检测及临床标本检测。 结果 针对人乳头瘤病毒58型所建立的实时荧光环介导等温扩增检测体系具有良好的特异性及灵敏度，且与临床实验室检测结果（qPCR方法）的符合率达100%，该检测方法的灵敏度可达10拷贝/μL。 结论 本研究建立了人乳头瘤病毒58型的实时荧光环介导等温扩增检测体系，它是一种耗时少、操作简便、特异性强、灵敏度高的方法，适用于HPV临床样本的快速检测。
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| 关 键 词： | 实时荧光环介导等温扩增技术 人乳头瘤病毒58型 快速检测 早期诊断 |
Detection of human papillomavirus type 58 gene based on real-time fluorescence loop mediated isothermal amplification |
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| HUANG Yue-ming,ZHOU Jun,MU Ying,BI Ying,ZHANG Zhi-qi,WU Qing-qing.Detection of human papillomavirus type 58 gene based on real-time fluorescence loop mediated isothermal amplification[J].Modern Preventive Medicine,2020,0(23):4326-4330. |
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| Authors: | HUANG Yue-ming ZHOU Jun MU Ying BI Ying ZHANG Zhi-qi WU Qing-qing |
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| Affiliation: | * Department of Microbiology and Immunology, Guizhou Medical University, Guiyang, Guizhou 550004, China |
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| Abstract: | Objective To establish a human papilloma type 58 gene detection system based on real-time fluorescent ring-mediated isothermal amplification for its rapid detection and early diagnosis. WTHZ]Methods Primer Explorer V5 was used to design primers for the conserved genes in the L1 region of HPV58. The nucleic acid dye SYBR Green Ⅰ was added before the reaction and the concentration of dNTPs, Mg2+ and internal primers (FIP, BIP) in the reaction system were optimized. The optimized concentrations were determined by real-time fluorescence amplification curve. WTHZ]Results The real-time fluorescence loop mediated isothermal amplification detection system established for human papillomavirus type 58 had excellent sensitivity, which was 10 copies/μL. The specificity of the assay was identical to that of qPCR. WTHZ]Conclusion This study establishes a real-time fluorescence loop mediated isothermal amplification detection system for human papillomavirus type 58, which is a less time, simple operation, highly specific and sensitive method, so is suitable for rapid detection of clinical samples of HPV. |
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| Keywords: | Real-time fluorescence loop mediated isothermal amplification technology Human papillomavirus type 58 Rapid detection Early diagnosis |
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