A rapid and highly sensitive protocol for the detection of Escherichia coli O157:H7 based on immunochromatography assay combined with the enrichment technique of immunomagnetic nanoparticles

基于免疫层析法结合免疫磁性纳米粒子富集技术的大肠杆菌O157:H7快速高灵敏检测方法

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作者:Hui Qi, Zhen Zhong, Han-Xin Zhou, Chun-Yan Deng, Hai Zhu, Jin-Feng Li, Xi-Li Wang, Fu-Rong Li

Background

Escherichia coli O157:H7 (E. coli O157:H7) is an important pathogenic bacterium that threatens human health. A rapid, simple, highly sensitive, and specific method for the detection of E. coli O157:H7 is necessary.

Conclusion

The kits have the potential to offer important social and economic benefits in the screening, monitoring, and control of food safety.

Methods

In the present study, immunomagnetic nanoparticles (IMPs) were prepared with nanopure iron as the core, coated with E. coli O157:H7 polyclonal antibodies. These IMPs were used in combination with immunochromatographic assay (ICA) and used to establish highly sensitive and rapid kits (IMPs+ICA) to detect E. coli O157:H7. The kits were then used to detect E. coli O157:H7 in 150 food samples and were compared with conventional ICA to evaluate their efficacy.

Results

The average diameter of IMPs was 56 nm and the amount of adsorbed antibodies was 106.0 μg/mg. The sensitivity of ICA and IMPs+ICA was 10(5) colony-forming units/mL and 10(3) CFUs/mL, respectively, for purified E. coli O157:H7 solution. The sensitivity of IMPs+ICA was increased by two orders, and its specificity was similar to ICA.

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