细菌活性检测方法研究进展及其应用探讨

刘丽; 魏强

doi:10.3784/jbjc.2021.000

细菌活性检测方法研究进展及其应用探讨

doi: 10.3784/jbjc.2021.000

刘丽,
魏强^,

中国疾病预防控制中心, 国家病原微生物资源库/国家病原微生物保藏中心北京 102206

基金项目: 国家重点研发计划 (2022YFC2602200)

详细信息

作者简介:
刘丽，女；山东省临沂市；研究生；病原微生物；Email：liuli12342022@163.com

魏强，男；吉林省延吉市；研究员；病原微生物；Email：weiqiang@chinacdc.cn

通讯作者:
魏强，Tel：010−58900338， Email：weiqiang@chinacdc.cn

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出版历程
- 收稿日期: 2023-09-22
- 网络出版日期: 2023-11-30

Research progress and application of bacterial activity detection methods

Liu Li,
Wei Qiang^,

National Pathogenic Microorganism Resource Bank / National Center for the Preservation of Pathogenic Microorganism, National Pathogen Resource Center Chinese Center for Diesae Control and Prevevtion, Beijing 102206, China

Funds: This study was supported by the National Key Research and Development Program of China (2022YFC2602200)

More Information

Corresponding author: Wei Qiang, Email: weiqiang@chinacdc.cn, Tel:0086-10-58900338

摘要

摘要: 细菌活性检测是检验、评价和保藏的关键，快速、灵敏、特异和准确的细菌活性检测方法是十分重要的。目前，细菌活性可以根据细菌生长繁殖能力、新陈代谢能力、细胞膜的完整性以及复制转录能力原理进行检测。不同的检测原理会有不同的检测方法，平板培养计数法是最为经典的活性检测方法，PMA-qPCR、荧光染料结合流式细胞仪、单细胞拉曼分析技术等是新应用的方法。随着科学技术的不断发展和细菌活性检测的需求，优化的细菌活性检测方法层出不穷，但不同的细菌活性检测方法具有不同的优缺点。本文就目前细菌活性检测原理、方法、应用以及优缺点等方面进行描述，并探讨了细菌活性检测方法在细菌检验和保藏等方面的应用。
- 细菌 /
- 活性 /
- 检测 /
- 评价 /
- 保藏
Abstract: Bacterial activity detection is the key to inspection, evaluation and preservation. Rapid, sensitive, specific and accurate bacterial activity detection methods are very important. At present, bacterial activity can be measured according to the principles of bacterial growth and reproduction ability, metabolic ability, membrane integrity, and replication and transcription ability. Different detection principles will have different detection methods, plate culture counting method is the most classic activity detection method, PMA-qPCR, fluorescent dye-combined flow cytometry, single-cell Raman analysis technology is the new application of instruments. With the continuous development of science and technology and the demand of bacterial activity detection, optimized bacterial activity detection methods are emerging in an endless stream, but different bacterial activity detection methods have different advantages and disadvantages. In this paper, the principle, method, application, advantages and disadvantages of bacterial activity detection were described, and the application of bacterial activity detection method in bacterial inspection and preservation was discussed.
- bacteria /
- Activity /
- Detection /
- Evaluate /
- preservation

HTML全文

表 1 细菌活性检测方法对比分析表

Table 1. Comparative analysis of bacterial activity detection methods

原理	方法	优点	不足
生长繁殖能力	平板培养计数法	1.低成本 2.可区分活菌和死菌 3.无需样本富集	1.所需时间较长（18-72h） 2.无法检测VBNC细菌，易造成假阴性结果
新陈代谢能力	ATP生物发光法	1.操作简单 2.检测时间短（1-5分钟）	1.提取ATP复杂 2.ATP易降解
	氧化还原法	1.操作简单	1.受浓度、温度等多种因素影响 2.指示剂有可能造成细胞毒性
	单细胞拉曼技术	1.无需培养细菌 2.无需标记细菌 3．快速、高效、自动化（整个过程只需要5h）	1.需建立数据库和专业分析软件 2.在细菌活性检测领域应用较少
	RT-PCR	1.灵敏度较高 2.特异性较高 3.自动化操作节省人力 4.可区分活菌和死菌	1.成本较高 2.mRNA的半衰期短，且易被降解，可造成假阴性结果
细胞膜的完整性和复制转录能力	PMA-qPCR	1.灵敏度较高 2.特异性较高 3.自动化操作节省人力 4.可区分活菌和死菌	1.无法完全去除死菌信号，可导致出现假阳性 2.无法区分具有完整细胞膜和无代谢活性的死菌
细胞膜的完整性	流式细胞仪	1.可计数VBNC细菌 2.可区分活菌和死菌 3.所需时间短（每秒达上万个单细胞或颗粒） 4.可提供详细细胞生理状态信息	1.需荧光染料标记，可能损害细菌活性 2.成本高 3.依赖大型仪器 4.需专业人员操作

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