霍乱弧菌核酸检测技术研究进展

Research progress on nucleic acid detection technology for Vibrio cholerae

中文关键词:  霍乱弧菌  核酸检测  聚合酶链式反应  等温扩增

英文关键词:Vibrio cholerae  nucleic acid detection  polymerase chain reaction  isothermal amplification

中文摘要:

      霍乱是由霍乱弧菌引起的烈性肠道传染病, 快速、准确识别病原体是疫情早期预警和应急处置的关键。本文系统综述近年来霍乱弧菌核酸检测技术的研究进展, 重点梳理聚合酶链式反应(polymerase chain reaction, PCR)技术和等温扩增技术的检测原理、技术特性与应用现状。通过构建核心性能综合对比表, 实现多维度横向评价; 深入剖析霍乱弧菌活菌不可培养状态检测、死菌DNA假阳性干扰、复杂基质抑制等当前领域关键共性问题, 并总结现有解决方案。结合临床诊断、现场应急、实验室精准检测等实际需求, 提出场景化技术选择建议: 实时荧光定量PCR技术为临床与实验室检测的金标准, 环介导等温扩增技术、重组酶聚合酶扩增技术适配现场快速检测, 微滴式数字PCR适用于精准定量及低载量样本确证。展望未来, 霍乱弧菌核酸检测将向“单氮化丙啶活菌鉴别+等温扩增+微流控集成+多靶标联检+智能判读”的一体化平台发展, 实现更快速、精准、便携的检测, 全面契合公共卫生防控实际需求。

英文摘要:

      Cholera is a severe intestinal infectious disease caused by Vibrio cholerae. Rapid and accurate identification of the pathogen is critical for early warning of outbreaks and emergency response. This review systematically examined recent advances in nucleic acid detection technologies for Vibrio cholerae, focused on the detection principles, technical characteristics and current applications of polymerase chain reaction (PCR) and isothermal amplification techniques, constructed a comprehensive performance comparison table to enable multi-dimensional comparative evaluation; thoroughly analyzed key common challenges in the field, including detection of viable but non-culturable Vibrio cholerae, false-positive interference from dead bacterial DNA, complex matrix inhibition and summarized existing solutions. Based on practical requirements in clinical diagnosis, on-site emergency testing and laboratory-based precision detection, this review proposed scenario-specific technical recommendations: real-time fluorescent quantitative PCR serves as the gold standard for clinical and laboratory testing; loop-mediated isothermal amplification and recombinase polymerase amplification were suitable for rapid on-site detection; and droplet digital PCR was applicable to precise quantification and confirmation of low-load samples. Looking ahead, Vibrio cholerae nucleic acid detection will evolve toward an integrated platform combining propidium monoazide-based viable bacteria discrimination, isothermal amplification, microfluidic integration, multi-target combined detection and intelligent interpretation, thereby enabling faster, more accurate and portable testing that fully meets the practical needs of public health prevention and control.