叠氮溴化丙锭染色技术在水体活性微生物监测中的应用

Application of PMA staining technique in monitoring viable microorganisms in aquatic ecosystems

摘要

【目的】叠氮溴化丙锭(PMA)染色技术作为一种选择性抑制死亡微生物DNA扩增的分子工具，近年来在水体活性微生物监测中展现出独特优势。传统分子检测方法无法区分死菌与活菌，导致结果不能反映出微生物真实的活性状态。PMA可以通过死亡细胞破损的细胞膜，与细胞内DNA结合，并阻断其被扩增，从而精准聚焦活性微生物群落的分析。【方法】系统总结了PMA染色技术相关的中英文文献，对比分析了其在水体活性微生物监测中优缺点。【结果】通过调节PMA浓度、优化光活化条件及联用qPCR或流式细胞术，显著提升了检测灵敏度和特异性。目前，该技术已成功应用于饮用水病原菌活体检测、污水处理、功能微生物追踪以及藻华活性微生物群落评估等领域。【结论】PMA技术为水体微生物研究提供了“生命状态”的观测工具，但其应用需结合具体场景优化，以期为饮用水安全、城市供水水质保障、水体生态修复及蓝藻治理提供更精准的技术支撑。

Abstract

[Objective] Propidium monoazide(PMA) staining technique, a molecular tool that selectively inhibits DNA amplification from nonviable microorganisms, has recently shown unique advantages in monitoring viable microorganisms in aquatic ecosystems. Conventional molecular detection method fail to distinguish between viable and nonviable bacteria, leading to result that cannot reflect the viability of microorganisms. PMA can permeate the compromised membranes of dead cells, bind to intracellular DNA, and block its amplification, thereby enabling targeted analysis of viable microbial communities.[Methods] A systematic review of Chinese and English literature related to PMA staining technique is conducted to comparatively analyze its advantages and disadvantages in monitoring viable microorganisms in aquatic ecosystems.[Results] By adjusting PMA concentration, optimizing photoactivation conditions, and integrating with qPCR or flow cytometry, the detection sensitivity and specificity are significantly improved. Currently, this technology has been successfully applied across diverse fields such as the detection of viable pathogens in drinking water, wastewater treatment, functional microorganism tracking, and the assessment of viable microbial communities in algal blooms.[Conclusion] PMA technology provides an effective tool for observing the “vital status” of aquatic microorganisms. However, its application requires optimization tailored to specific scenarios to provide more precise technical support for drinking water safety, urban water supply quality assurance, aquatic ecosystem restoration, and cyanobacterial bloom management.

关键词

叠氮溴化丙锭 / 水生态 / 活性微生物 / 环境DNA / 染色技术 / 微生物监测 / 饮用水安全 / 水生态修复

Key words

propidium monoazide / aquatic ecosystem / viable microorganisms / environmental DNA / staining techniques / restoration microbiac monitoning / drinking water safety / water ecological

基金资助

国家自然科学基金项目(32201352)

温州市科学技术局基础性公益科研项目(S20240011)