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应用 PMA 辅助 qPCR 从饮用水和水源水中检测活的沙门氏菌。

Determination of viable Salmonellae from potable and source water through PMA assisted qPCR.

机构信息

Environmental Microbiology, Environmental Toxicology Group, CSIR-Indian Institute of Toxicology Research, Post Box 80, Mahatma Gandhi Marg, Lucknow-226001, Uttar Pradesh, India.

出版信息

Ecotoxicol Environ Saf. 2013 Jul;93:121-7. doi: 10.1016/j.ecoenv.2013.02.017. Epub 2013 Apr 26.

Abstract

Resource constrained countries identified as endemic zones for pathogenicity of Salmonella bear an economic burden due to recurring expenditure on medical treatment. qPCR used for Salmonella detection could not discriminate between viable and nonviable cells. Propidium monoazide (PMA) that selectively penetrates nonviable cells to cross-link their DNA, was coupled with ttr gene specific qPCR for quantifying viable salmonellae in source/potable waters collected from a north Indian city. Source water (raw water for urban potable water supply) and urban potable water exhibited viable salmonellae in the range of 2.1×10(4)-2.6×10(6) and 2-7160CFU/100mL, respectively. Potable water at water works exhibited DNA from dead cells but no viable cells were detected. PMA assisted qPCR could specifically detect low numbers of live salmonellae in Source and potable waters. This strategy can be used in surveillance of urban potable water distribution networks to map contamination points for better microbial risk management.

摘要

资源有限的国家被确定为沙门氏菌致病性的流行地区,由于需要经常支出医疗费用,因此承担着经济负担。用于沙门氏菌检测的 qPCR 无法区分活细胞和非活细胞。吖啶橙(PMA)选择性穿透非活细胞以交联其 DNA,与 ttr 基因特异性 qPCR 结合,用于定量来自印度北部城市的水源/饮用水中存活的沙门氏菌。水源(城市饮用水供应的原水)和城市饮用水中分别存在活的沙门氏菌,范围为 2.1×10(4)-2.6×10(6)和 2-7160CFU/100mL。水厂的饮用水仅显示死细胞的 DNA,但未检测到活细胞。PMA 辅助 qPCR 可以特异性检测水源和饮用水中低数量的活沙门氏菌。该策略可用于监测城市饮用水分配网络,以确定污染点,从而更好地进行微生物风险管理。

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