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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 .    Electronic supplementary material  The online version of this article (10.1186/s12866-018-1241-5) contains supplementary material, which is available to authorized users.

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