The present paper aims to study the sensitivity and specificity of loop-mediated isothermal amplification (LAMP) in detection of Yersinia pestis (Y. pestis) from all natural plague foci of China. The LAMP was established based on the specific primers targeting 3a gene of Y. pestis. Sixty-five Y. pestis strains were isolated from 11 natural plague foci of China, and Y. pseudotuberculosis, Y. enterocolitica and Escherichia coli (E. coli) were set as control strains. The results of LAMP detection were consistent with three standard diagnostic methods of plague (reverse indirect haemagglutination test, colloidal gold immunochromatography assay and conventional polymerase chain reaction method). The 65 Y. pestis strains were all positive, while Y. pseudotuberculosis, Y. enterocolitica and E. coli were negative, suggesting that the established LAMP method was highly specific for Y. pestis. It has the advantages of fast, simple, low requirements on equipment, with a potential application in field monitoring and detection in remote areas for plague prevention and control.