Related Experiment Video

Updated: Jul 3, 2026

Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis
10:33

Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis

Published on: June 17, 2019

Development of a real-time quantitative PCR assay to enumerate Yersinia pestis in fleas

Elizabeth S Gabitzsch1, Rommelle Vera-Tudela, Rebecca J Eisen

  • 1Centers for Disease Control and Prevention, Division of Vector-Borne Infectious Diseases, Fort Collins, Colorado 80521, USA.

The American Journal of Tropical Medicine and Hygiene
|July 9, 2008
PubMed

Insights

A new real-time quantitative polymerase chain reaction (qPCR) assay accurately detects Yersinia pestis. This rapid method allows for quick identification and counting of Yersinia pestis bacteria in fleas.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Infectious Diseases

Background:

  • Yersinia pestis is the causative agent of plague.
  • Accurate and rapid detection methods for Yersinia pestis are crucial for disease control.
  • Conventional methods for bacterial enumeration can be time-consuming.

Purpose of the Study:

  • To develop and optimize a real-time quantitative polymerase chain reaction (qPCR) assay for the detection and enumeration of Yersinia pestis.
  • To validate the assay's performance using cultured strains and infected flea samples.

Main Methods:

  • Design of primers and a TaqMan probe targeting a conserved region of the Yersinia pestis ferric iron uptake regulator (fur) gene.
  • Optimization of the qPCR assay using cultured Yersinia pestis.
  • Validation of the assay with multiple Yersinia pestis biovars and in infected flea triturates.

Main Results:

  • The developed qPCR assay successfully detected Yersinia pestis.
  • The assay demonstrated high sensitivity, detecting as few as 300 bacteria in infected flea triturates.
  • The assay was confirmed to be effective across three Yersinia pestis biovars.

Conclusions:

  • A sensitive and specific real-time qPCR assay for Yersinia pestis has been established.
  • This assay enables rapid and accurate enumeration of Yersinia pestis in laboratory settings, particularly in infected fleas.
  • The qPCR method offers a significant advantage over traditional culture-based techniques for Yersinia pestis quantification.

More Related Videos

'Bioluminescent' Reporter Phage for the Detection of Category A Bacterial Pathogens11:31

'Bioluminescent' Reporter Phage for the Detection of Category A Bacterial Pathogens

Published on: July 8, 2011

Detecting the Lyme Disease Spirochete, Borrelia Burgdorferi, in Ticks Using Nested PCR07:20

Detecting the Lyme Disease Spirochete, Borrelia Burgdorferi, in Ticks Using Nested PCR

Published on: February 4, 2018

Related Concept Videos

Plague01:24

Plague

Plague is a highly virulent zoonotic disease caused by Yersinia pestis, a Gram-negative, facultatively anaerobic coccobacillus. This pathogen primarily circulates among rodent populations and is transmitted to humans through the bite of infected fleas. Additional transmission routes include direct contact with infected animal tissue or inhalation of respiratory droplets from individuals with pneumonic plague. These multiple transmission pathways highlight the bacterium’s potential for rapid...
Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...