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

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.

A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting the pathogen of maize rough dwarf disease in China

“…The results of the reaction can be observed by adding a fluorescent indicator calcein or through the direct visualization of precipitation in the tubes (Tomita et al 2008;Mori et al 2001). RT-LAMP was widely applied for the detection of pathogens (Keizerweerd et al 2015;Hu et al 2016;Yoshikawa et al 2014;Gao et al 2015;Nair et al 2016). Here, the RT-LAMP assay was developed for rapid and sensitive detection of the MRDD pathogen of maize and the small brown planthopper.…”

groEL is a suitable genetic marker for detecting Vibrio parahaemolyticus by loop-mediated isothermal amplification assay

“…b), similar to the findings of Hu et al . (), who reported a LoD of 125 fg per LAMP reaction for detecting B. quintana using the groEL gene. Zhao et al .…”

“…) and Bartonella quintana (Hu et al . ), and the groESL operon for Ehrlichia canis (Faggion et al . ).…”