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微生物灭活以安全快速诊断感染性样本。

Microbial inactivation for safe and rapid diagnostics of infectious samples.

机构信息

Wehrwissenschaftliches Institut für Schutztechnologien ABC-Schutz, Munster, Germany.

出版信息

Appl Environ Microbiol. 2011 Oct;77(20):7289-95. doi: 10.1128/AEM.05553-11. Epub 2011 Aug 19.

Abstract

The high risk associated with biological threat agents dictates that any suspicious sample be handled under strict surety and safety controls and processed under high-level containment in specialized laboratories. This study attempted to find a rapid, reliable, and simple method for the complete inactivation of a wide range of pathogens, including spores, vegetative bacteria, and viruses, while preserving microbial nucleic acid fragments suitable for PCRs and proteinaceous epitopes for detection by immunoassays. Formaldehyde, hydrogen peroxide, and guanidium thiocyanate did not completely inactivate high titers of bacterial spores or viruses after 30 min at 21°C. Glutaraldehyde and sodium hypochlorite showed high microbicidal activity but obliterated the PCR or enzyme-linked immunosorbent assay (ELISA) detection of bacterial spores or viruses. High-level inactivation (more than 6 log(10)) of bacterial spores (Bacillus atrophaeus), vegetative bacteria (Pseudomonas aeruginosa), an RNA virus (the alphavirus Pixuna virus), or a DNA virus (the orthopoxvirus vaccinia virus) was attained within 30 min at 21°C by treatment with either peracetic acid or cupric ascorbate with minimal hindrance of subsequent PCR tests and immunoassays. The data described here should provide the basis for quickly rendering field samples noninfectious for further analysis under lower-level containment and considerably lower cost.

摘要

与生物威胁剂相关的高风险决定了任何可疑样本都必须在严格的安全保障和控制下进行处理,并在专门的实验室中进行高水平隔离处理。本研究试图找到一种快速、可靠且简单的方法,以完全灭活广泛的病原体,包括孢子、营养细菌和病毒,同时保留适合 PCR 的微生物核酸片段和用于免疫测定的蛋白质表位。甲醛、过氧化氢和硫氰酸胍在 21°C 下 30 分钟后不能完全灭活高滴度的细菌孢子或病毒。戊二醛和次氯酸钠显示出很高的杀菌活性,但会破坏细菌孢子或病毒的 PCR 或酶联免疫吸附测定(ELISA)检测。在 21°C 下 30 分钟内,通过用过氧乙酸或抗坏血酸铜处理,可以实现细菌孢子(萎缩芽孢杆菌)、营养细菌(铜绿假单胞菌)、RNA 病毒(阿尔法病毒皮库纳病毒)或 DNA 病毒(正痘病毒牛痘病毒)的高水平灭活(超过 6 个对数),对随后的 PCR 检测和免疫测定几乎没有阻碍。这里描述的数据应该为快速使现场样本失去传染性以便在较低水平的隔离和成本下进行进一步分析提供基础。

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