Characterization of an F1 deletion mutant of Yersinia pestis CO92, pathogenic role of F1 antigen in bubonic and pneumonic plague, and evaluation of sensitivity and specificity of F1 antigen capture-based dipsticks

J Clin Microbiol. 2011 May;49(5):1708-15. doi: 10.1128/JCM.00064-11. Epub 2011 Mar 2.

Abstract

We evaluated two commercial F1 antigen capture-based immunochromatographic dipsticks, Yersinia Pestis (F1) Smart II and Plague BioThreat Alert test strips, in detecting plague bacilli by using whole-blood samples from mice experimentally infected with Yersinia pestis CO92. To assess the specificities of these dipsticks, an in-frame F1-deficient mutant of CO92 (Δcaf) was generated by homologous recombination and used as a negative control. Based on genetic, antigenic/immunologic, and electron microscopic analyses, the Δcaf mutant was devoid of a capsule. The growth rate of the Δcaf mutant generally was similar to that of the wild-type (WT) bacterium at both 26 and 37 °C, although the mutant's growth dropped slightly during the late phase at 37 °C. The Δcaf mutant was as virulent as WT CO92 in the pneumonic plague mouse model; however, it was attenuated in developing bubonic plague. Both dipsticks had similar sensitivities, requiring a minimum of 0.5 μg/ml of purified F1 antigen or 1 × 10(5) to 5 × 10(5) CFU/ml of WT CO92 for positive results, while the blood samples were negative for up to 1 × 10(8) CFU/ml of the Δcaf mutant. Our studies demonstrated the diagnostic potential of two plague dipsticks in detecting capsular-positive strains of Y. pestis in bubonic and pneumonic plague.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bacterial Proteins / analysis*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Clinical Laboratory Techniques / methods*
  • Disease Models, Animal
  • Female
  • Gene Deletion*
  • Immunoassay
  • Mice
  • Mice, Inbred BALB C
  • Plague / diagnosis*
  • Plague / microbiology
  • Plague / mortality
  • Plague / pathology*
  • Reagent Kits, Diagnostic
  • Sensitivity and Specificity
  • Survival Analysis
  • Virulence
  • Virulence Factors / genetics*
  • Virulence Factors / metabolism
  • Yersinia pestis / genetics
  • Yersinia pestis / growth & development
  • Yersinia pestis / pathogenicity*

Substances

  • Bacterial Proteins
  • Reagent Kits, Diagnostic
  • Virulence Factors
  • caf1 protein, Yersinia pestis