Rapid, Sensitive Detection of Bartonella quintana by Loop-Mediated Isothermal Amplification of the groEL Gene

Int J Mol Sci. 2016 Dec 1;17(12):1902. doi: 10.3390/ijms17121902.

Abstract

Trench fever, caused by Bartonella quintana, is recognized as a re-emerging and neglected disease. Rapid and sensitive detection approaches are urgently required to monitor and help control B. quintana infections. Here, loop-mediated isothermal amplification (LAMP), which amplifies target DNA at a fixed temperature with high sensitivity, specificity and rapidity, was employed to detect B. quintana. Thirty-six strains, including 10 B. quintana, 13 other Bartonella spp., and 13 other common pathogens, were applied to verify and evaluate the LAMP assay. The specificity of the LAMP assay was 100%, and the limit of detection was 125 fg/reaction. The LAMP assay was compared with qPCR in the examination of 100 rhesus and 20 rhesus-feeder blood samples; the diagnostic accuracy was found to be 100% when LAMP was compared to qPCR, but the LAMP assay was significantly more sensitive (p < 0.05). Thus, LAMP methodology is a useful for diagnosis of trench fever in humans and primates, especially in low-resource settings, because of its rapid, sensitive detection that does not require sophisticated equipment.

Keywords: Bartonella; detection; re-emerging pathogen; trench fever.

MeSH terms

  • Animals
  • Bartonella quintana / genetics
  • Bartonella quintana / isolation & purification*
  • Bartonella quintana / pathogenicity
  • Chaperonin 60 / blood*
  • Chaperonin 60 / genetics
  • Humans
  • Macaca mulatta / blood
  • Macaca mulatta / microbiology
  • Real-Time Polymerase Chain Reaction / methods
  • Trench Fever / blood*
  • Trench Fever / genetics
  • Trench Fever / microbiology

Substances

  • Chaperonin 60