Abstract
BackgroundCoxiella burnetii is an obligate intracellular Gram-negative bacterium that causes a zoonotic disease commonly called Q fever globally. In this study, an up-converting phosphor technology-based lateral flow (UPT-LF) assay was established for the rapid and specific detection of phase I strains of C. burnetii.ResultsSpecific monoclonal antibodies (10B5 and 10G7) against C. burnetii phase I strains were prepared and selected for use in the UPT-LF assay by the double-antibody-sandwich method. The detection sensitivity of the Coxiella-UPT-LF was 5 × 104 GE/ml for a purified C. burnetii phase I strain and 10 ng/ml for LPS of C. burnetii Nine Mile phase I (NMI). Good linearity was observed for C. burnetii phase I and NMI LPS quantification (R2 ≥ 0.989). The UPT-LF assay also exhibited a high specificity to C. burnetii, without false-positive results even at 108 GE/ml of non-specific bacteria, and good inclusivity for detecting different phase I strains of C. burnetii. Moreover, the performance of the Coxiella-UPT-LF assay was further confirmed using experimentally and naturally infected samples.ConclusionsOur results indicate that Coxiella-UPT-LF is a sensitive and reliable method for rapid screening of C. burnetii, suitable for on-site detection in the field.
Highlights
Coxiella burnetii is an obligate intracellular Gram-negative bacterium that causes a zoonotic disease commonly called Q fever globally
Development of Coxiella-up-converting phosphor technology-based lateral flow (UPT-LF) The monoclonal antibodies against C. burnetii were prepared in mice that were immunised with purified C. burnetii Xinqiao strain (PI)
Three cloned hybridomas (10B5, 10G7, and 13D6) that produced C. burnetii Phase I (PI)-specific monoclonal antibodies (mAbs) and two cloned hybridomas (6D8 and 8A1) that produced both PI- and Phase II (PII)-specific mAb were identified by ELISA analysis of hybridoma supernatants with PI and PII antigens
Summary
Coxiella burnetii is an obligate intracellular Gram-negative bacterium that causes a zoonotic disease commonly called Q fever globally. Coxiella burnetii is an intracellular Gram-negative bacterium that causes a zoonotic disease known as Q fever globally. It can undergo a phase transition that is correlated with some of the biological characteristics of the “smooth-to-rough” lipopolysaccharide (LPS) variation observed for Gram-negative Enterobacteriaceae [1]. Upon serially passaging in embryonic cells, tissue culture, or synthetic medium, a smooth-torough (truncated) LPS transition occurs, which results in avirulence (phase II, PII) [3]. Diagnosis of Q fever is difficult due to the lack of distinct clinical features that distinguish it from other febrile diseases [5]. The diagnosis of Q fever mainly depends on the detection of antibodies or nucleic
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