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?:abstract
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Glaesserella parasuis is a fastidious pathogen that colonizes the respiratory tract of pigs and can lead to considerable economic losses in pig production Therefore, a rapid detection assay for the pathogen, preferably applicable in the field, is important In the current study, we developed a new and improved detection method using loop-mediated isothermal amplification (LAMP) This assay, which targets the infB gene, was tested on a collection of 60 field isolates of G parasuis comprising 14 different serovars In addition, 63 isolates from seven different closely related species of the family Pasteurellaceae, including A indolicus, A porcinus, and A minor, and a species frequently found in the respiratory tract of pigs were used for exclusivity experiments This assay showed an analytical specificity of 100% (both inclusivity and exclusivity) and an analytical sensitivity of 10 fg/µL In further steps, 36 clinical samples were tested with the LAMP assay An agreement of 77 1 (95% CI: 59 9, 89 6) and 91 4% (95% CI: 75 9, 98 2) to the culture-based and PCR results was achieved The mean limit of detection for the spiked bronchoalveolar lavage fluid was 2 58 × 102 CFU/mL A colorimetric assay with visual detection by the naked eye was tested to provide an alternative method in the field and showed the same sensitivity as the fluorescence-based LAMP assay Overall, the optimized LAMP assay represents a fast and reliable method and is suitable for detecting G parasuis in the laboratory environment or in the field
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