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Rapid and sensitive detection of Yersinia pestis by lateral-flow assay in simulated clinical samples.
Hsu, Hui-Ling; Chuang, Chuan-Chang; Liang, Chung-Chih; Chiao, Der-Jiang; Wu, Hsueh-Ling; Wu, Yu-Ping; Lin, Feng-Ping; Shyu, Rong-Hwa.
Afiliación
  • Hsu HL; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan.
  • Chuang CC; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan.
  • Liang CC; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan.
  • Chiao DJ; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan.
  • Wu HL; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan.
  • Wu YP; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan.
  • Lin FP; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan.
  • Shyu RH; Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, Taipei, Taiwan. shyu11@yahoo.com.tw.
BMC Infect Dis ; 18(1): 402, 2018 08 14.
Article en En | MEDLINE | ID: mdl-30107826
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 103 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.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteínas Bacterianas / Yersinia pestis / Inmunoensayo Tipo de estudio: Diagnostic_studies Límite: Animals / Humans Idioma: En Revista: BMC Infect Dis Asunto de la revista: DOENCAS TRANSMISSIVEIS Año: 2018 Tipo del documento: Article País de afiliación: Taiwán

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteínas Bacterianas / Yersinia pestis / Inmunoensayo Tipo de estudio: Diagnostic_studies Límite: Animals / Humans Idioma: En Revista: BMC Infect Dis Asunto de la revista: DOENCAS TRANSMISSIVEIS Año: 2018 Tipo del documento: Article País de afiliación: Taiwán