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Examining the vector-host-pathogen interface with quantitative molecular tools

Jason E Comer1, Ellen A Lorange, B Joseph Hinnebusch

  • 1Plague Section, Laboratory of Zoonotic Pathogens, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 22, 2008
PubMed

Insights

New PCR assays accurately detect and quantify Yersinia pestis, the plague bacterium, in fleas and animal tissues. These methods aid in understanding plague transmission dynamics in arthropod-borne diseases.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Epidemiology

Background:

  • Plague is a serious zoonotic disease caused by Yersinia pestis.
  • Effective detection and quantification methods are crucial for understanding plague transmission dynamics.

Purpose of the Study:

  • To develop and validate Polymerase Chain Reaction (PCR) assays for detecting and quantifying Yersinia pestis.
  • To apply these assays to study plague transmission in vector and host tissues.

Main Methods:

  • Developed real-time PCR assays using primers and probes specific to a multi-copy plasmid of Y. pestis.
  • Quantified bacterial numbers in fleas, fleabite sites, and lymph nodes.
  • Utilized tissue-matched standard curves for accurate absolute bacterial quantification.

Main Results:

  • The PCR assays demonstrated high specificity and sensitivity for Yersinia pestis detection.
  • Linear quantification was achieved over at least five orders of magnitude.
  • The methods were successfully applied to studies involving Xenopsylla cheopis flea transmission.

Conclusions:

  • The developed PCR assays provide a reliable tool for detecting and quantifying Yersinia pestis.
  • These methods are valuable for investigating the transmission dynamics of plague and potentially other arthropod-borne diseases.

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