Current Situation and Prospect of New Techniques for Rapid Clinical Microbiological Testing
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摘要: 感染性疾病起病急、进展快,早期精准识别和监测病原体耐药性对患者预后及遏制耐药至关重要。临床微生物常规技术已无法满足快速诊疗的需求,因此快速检测技术成为检验与临床关注的焦点。本文论述快速鉴定与药物敏感性检测的最新技术研究现状、问题及未来发展要点,旨在为临床微生物实验室未来新技术的引入提供参考。
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关键词:
- 临床微生物学 /
- 快速检测 /
- 鉴定 /
- 基质辅助激光解吸电离飞行时间质谱 /
- 光谱技术 /
- 电化学生物传感器 /
- 宏基因组测序 /
- 体外抗菌药物敏感性试验
Abstract: Infectious diseases begin and progress rapidly, so early accurate identification and detection of pathogen resistance is crucial for the prognosis of patients and the curb on drug resistance. Conventional clinical microbiological technology has been unable to meet the needs of rapid diagnosis and treatment. Therefore, rapid detection technology becomes the focus of clinic and clinical laboratory. This paper reviews the research status of the latest technology for rapid identification and detection of drug sensitivity, and discusses their problems and the key points of future development, providing reference to the introduction of new technologies in the clinical microbiological laboratory in the future.作者贡献:宁雅婷负责撰写、修订文章;杨启文、陈新飞、郁谨菡、李雪负责收集并整理文献;徐英春负责审校文章。利益冲突:无 -
表 1 不同光谱技术原理及其在临床微生物检测领域的应用
名称 原理 用途 现状或应用前景 主要优缺点 拉曼光谱 (1)对散射光谱进行分析,得到生物大分子结构特征振动信息图谱;记录大分子结构组成变化,结合多元统计分析区分不同种属的细菌 分类鉴定 鉴定系统仍然处于开发阶段,亟需搭建光谱数据库
可在十几秒内完成对葡萄球菌、单核细胞增生性李斯特菌快速区分检测[11];40 min内完成血清样本中念珠菌种水平上的鉴定[12];结合化学计量学方法实现链球菌快速区分[13]等优点:不受水分子干扰,非常适合生物医学体系;不依赖增殖培养,实现准确、高通量、对微生物无损伤、超快检[14];实时动态监测生物大分子含量,获得丰富的生物信息
缺点:易受荧光信号干扰,重现性不佳,信号弱,设备价格高昂(2)结合重水同位素标记检测细胞代谢活性[15];检测细胞运动,以判断细菌活力 耐药性检测 用于发现潜在耐药性;监测医院内病原菌的流行病学;区分抗菌药物处理和未经处理的细菌,并在3.5 h内精确识别肠球菌中耐万古霉素菌株[16] (3)结合其他单细胞分离技术分离菌株,如光镊技术 菌株分离 对于混合菌群、苛养菌、异质性细菌研究具有深远意义
微流控芯片结合光镊技术可分离出单细胞大肠埃希菌[16]近红外光谱 借助红外光和生物大分子化学键作用所产生的合频及振动倍频,获得微生物含氢基团的特征信息 快速鉴定 尚未用于临床,近红外光穿透性高,在深层组织分析应用方面极具前景[17] 优点:抗荧光干扰力强,可定性和定量分析
缺点:灵敏度稍低,建模难度大,易受水分子干扰高光谱图像 遥感技术,精确识别化学组分的微小变化(光谱信息),反映微生物外部多层次的变化(图像信息)[18] 快速鉴定 一种新型的、非接触式的光学诊断技术,为临床提供有效的辅助诊断手段,具有巨大的发展潜力 优点:可将影像与光谱信息密切结合,解决光谱无法成像的瓶颈
缺点:空间分辨率、信噪比稍差激光诱导击穿光谱 利用激光照射微生物表面产生等离子体,探测等离子体中的原子和离子谱线 快速鉴定 处于研究起步阶段
真菌方面研究及深度远不及细菌
采用纳米颗粒与横向流带结合构成新的传感平台激光诱导击穿光谱横向流带,在10 min内可对金黄色葡萄球菌进行检测,检出下限达1.6 CFU/mL[19]优点:实时在线、非接触、多种元素同时探测
缺点:在检测灵敏度、消除基质效应及便携等方面存在挑战 -
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