Development and evaluation of an up-converting phosphor technology-based lateral flow assay for rapid and quantitative detection of Coxiella burnetii phase I strains

Abstract

Background: Coxiella 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. Results: Specific 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 × 10 4 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 (R 2 ≥ 0.989). The UPT-LF assay also exhibited a high specificity to C. burnetii, without false-positive results even at 10 8 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. Conclusions: Our 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.

A protein–protein interaction map reveals that the Coxiella burnetii effector CirB inhibits host proteasome activity

“…Because virulent C . burnetii phase I strains have full-length LPS and can successfully infect and propagate in immunocompetent mice [ 50 ], C57BL/6J-wild-type (WT) and C57BL/6J- psmb5 +/- mice ( S8D Fig ) were used to study the relationship between PSMB5 function and the replication of C . burnetii virulent strains.…”

An up-converting phosphor technology-based lateral flow assay for rapid detection of major mycotoxins in feed: Comparison with enzyme-linked immunosorbent assay and high-performance liquid chromatography-tandem mass spectrometry

“…Compared with the HPLC and ELISA methods, the UPT-LF method is quicker and easier to operate. Rapid detection is one of the obvious advantages of UPT-LF method which has been reported in numerous researches [ 21 , 31 , 32 ].…”

“…Combining UCP with a lateral flow immunoassay exhibits little background interference from complex matrices, which ensures its high sensitivity and stability. To date, the UPT-LF assay has been widely used for the sensitive detection of drugs of abuse [ 18 ], nucleic acids [ 19 ], interferon-γ [ 20 ], and various infectious pathogens and foodborne pathogens, such as Escherichia coli [ 18 ], Coxiella burnetii [ 21 ] and Yersinia pestis [ 22 ].…”

Minimizing Cross-Reactivity for the Chemiluminescent Lateral Flow Immunoassay of Cardiac Troponin I Based on PEGylation of Gold Nanoparticles

“…Great efforts have been made to diminish the false-positive errors and improve the detection accuracy. − One of the effective methods is to remove the Fc region of the antibody, which is the main reason for the nonspecificity . In addition to it, biotinylated antibodies can also be applied .…”

“…The hydrophobic cadmium-based quantum dots with enhanced dispersibility were studied as the probe for detecting SARS-CoV-2 that can minimize the cross-reactivity and increase the accuracy . Other nanoparticles, such AuNPs, up-converting NPs, and magnetic iron-oxide NPs, , have also been used to detect the biomarkers with high accuracy and sensitivity. For example, Peterson et al utilized antibody-functionalized magnetic iron-oxide nanoparticles to decrease the nonspecific signals and detect the biomarkers with high accuracy.…”