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利用暗场显微镜下的磁性纳米颗粒探针实现大肠杆菌的超灵敏快速计数。

Ultrasensitive and rapid count of Escherichia coli using magnetic nanoparticle probe under dark-field microscope.

机构信息

Institute of Comparative Medicine, Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, Yangzhou, 225009, China.

Key Laboratory of Animal Bacteriology, Ministry of Agriculture, College of veterinary medicine, Nanjing Agricultural University, Nanjing, 210095, China.

出版信息

BMC Microbiol. 2018 Sep 3;18(1):100. doi: 10.1186/s12866-018-1241-5.

Abstract

BACKGROUND

Escherichia coli (E. coli) is one of the best-known zoonotic bacterial species, which pathogenic strain can cause infections in humans and animals. However, existing technologies or methods are deficient for quickly on-site identifying infection of E. coli before they breakout. Herein, we present an ultrasensitive and on-site method for counting E. coli using magnetic nanoparticle (MNP) probe under a dark-field in 30 min.

RESULTS

The antibodies functionalized MNP, binding to E. coli to form a golden ring-like structure under a dark-field microscope, allowing for counting E. coli. This method via counting MNP-conjugated E. coli under dark-field microscope demonstrated the sensitivity of 6 CFU/μL for E. coli detection. Importantly, due to the advantages such as time-saving (only 30 min) and almost free of instrument (only require a portable microscope), our MNP-labeled dark-field counting strategy has the potential of being a universal tool for on-site quantifying a variety of pathogens with size ranges from a few hundreds of nanometers to a few micrometers.

CONCLUSION

In summary, the MNP-labeled dark-field counting strategy is a rapid, simple, sensitive as well as low-cost assay strategy, which has the potential of being a universal tool for on-site quantification of micrometer-size pathogens like E. coli.

摘要

背景

大肠杆菌(E. coli)是最知名的人畜共患病细菌物种之一,其致病菌株可引起人类和动物的感染。然而,现有的技术或方法在其爆发之前无法快速现场识别大肠杆菌的感染。在此,我们提出了一种在 30 分钟内使用暗场下磁性纳米颗粒(MNP)探针超灵敏和现场计数大肠杆菌的方法。

结果

抗体功能化的 MNP 在暗场显微镜下与大肠杆菌结合形成金环状结构,从而可以计数大肠杆菌。该方法通过在暗场显微镜下计数 MNP 结合的大肠杆菌,检测大肠杆菌的灵敏度为 6 CFU/μL。重要的是,由于节省时间(仅 30 分钟)和几乎无需仪器(仅需要便携式显微镜)等优点,我们的 MNP 标记暗场计数策略有可能成为现场定量分析从几百纳米到几微米大小的各种病原体的通用工具。

结论

总之,MNP 标记暗场计数策略是一种快速、简单、灵敏且低成本的检测策略,有可能成为现场定量分析大肠杆菌等微米大小病原体的通用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00e3/6122661/8c0b6ea90a07/12866_2018_1241_Fig1_HTML.jpg

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