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Background   Yersinia pestis  is a contributing agent to the epidemic disease, plague, which killed an estimated 200 million people during historical times. In this study, a rapid, cheap, sensitive, and specific technique, the lateral flow assay (F1 strips), has been successfully developed to detect this pathogen, by using paired monoclonal antibodies (MAbs) against  Y. pestis  capsule like fraction 1 (F1) protein. Compared with the polyclonal antibody (PAb) based F1 strips, the Mab-based F1 strips have a remarkable increased detection limitation (10 to 100 folds). Furthermore, besides the limitation and specificity evaluation, the application of this F1 strip on simulated clinical samples indicate the LFA can be a good candidate to detect plague.    Methods  Recombinant F1 antigen was expressed and purified from a series of works. The various anti-F1 monoclonal antibodies generated from hybridoma cells were screened with the ELISA technique. To evaluate the feasibility of this  Y. pestis  F1 test strip, the F1 protein/ Y. pestis  was spiked into simulated clinical samples such as human serum, mouse bronchoalveolar lavage fluids, and mouse blood to mimic natural infection status. Additionally, this technique was applied to detect the  Y. pestis  in the environment-captured rats, to evaluate the practical usefulness of the strips.    Results  By using this MAb-based-LFA technique, 4 ng/ml of recombinant F1-protein and 10 3  CFU/ml of  Y. pestis  could be detected in less than 10 mins, which is at least 10-folds than that of the PAb format. On the other hand, although various  Yersinia  strains were applied to the strips, only  Y. pestis  strain showed a positive result; all other  Yersinia  species did not produce a positive signal, indicating the high efficiency and specificity of the MAb-based F1-strips.    Conclusion  Based on our findings, we suggest that the MAb-format-LFA will be valuable as a diagnostic tool for the detection of  Y. pestis . This report shows that the F1 strip is sufficient to support not only the detection of plague in simulated clinical samples, but also it may be a good candidate to meet the epidemiological surveillance during an outbreak of the biological warfare.

Rapid and sensitive detection of Yersinia pestis by lateral-flow assay in simulated clinical samples