Grimaldo Miguel A, Mendell Nicole L, Bopp Nathen E, Bouyer Donald H, Mafra Claudio
Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brasil.
Department of Pathology, University of Texas Medical Branch, Galveston, Texas, USA.
Appl Biosaf. 2021 Sep 1;26(3):130-138. doi: 10.1089/apb.20.0060. Epub 2021 Sep 13.
Ionized hydrogen peroxide (iHP) is a new technology used for the decontamination of surfaces or laboratory areas. It utilizes a low concentration of hydrogen peroxide (HO) mixed with air and ionized through a cold plasma arc. This technology generates reactive oxygen species as a means of decontamination. The purpose of this study is to review the effects of iHP on the structure of the spores of by observing its effects using transmission electron microscopy (TEM) and also by evaluating the existence of DNA damage by fluorescence-based quantitative polymerase chain reaction (qPCR). Spore samples of decontaminated using iHP at different exposure times (Control, 1, 2, 6, and 12 h) were fixed for TEM. In addition, DNA was extracted for evaluation of DNA damages using fluorescence-based qPCR assays. Damages to the spore structures of caused by the decontamination process with iHP at different exposure times (Control, 1, 2, 6, and 12 h) can be observed in micrographs. The effects of the decontamination to short DNA segment (132 base pairs [bp]) of the gene using qPCR present a linear degradation, and for the long DNA segment (680 bp), it presents a biphasic mode. The results of the qPCR analysis show two initial stages of damage to DNA with very noticeable damage at 12 h contact time, which confirms the observations of the TEM micrographs for the spores. The study demonstrates damage to the spore core DNA.
离子化过氧化氢(iHP)是一种用于表面或实验室区域去污的新技术。它利用低浓度的过氧化氢(HO)与空气混合,并通过冷等离子体弧进行离子化。该技术通过产生活性氧物质来实现去污目的。本研究的目的是通过透射电子显微镜(TEM)观察iHP对[具体菌种]孢子结构的影响,并通过基于荧光的定量聚合酶链反应(qPCR)评估DNA损伤的存在情况。使用iHP在不同暴露时间(对照、1、2、6和12小时)对[具体菌种]的孢子样本进行去污处理后,固定用于TEM观察。此外,提取DNA,使用基于荧光的qPCR分析评估DNA损伤情况。在显微照片中可以观察到,在不同暴露时间(对照、1、2、6和12小时)下,iHP去污过程对[具体菌种]孢子结构造成的损伤。使用qPCR对[具体菌种]基因的短DNA片段(132个碱基对[bp])进行去污处理的效果呈现线性降解,而对于长DNA片段(680 bp),则呈现双相模式。qPCR分析结果显示,DNA损伤有两个初始阶段,在接触12小时时损伤非常明显,这证实了对[具体菌种]孢子TEM显微照片的观察结果。该研究证明了孢子核心DNA受到损伤。