Rapid detection of tetracycline resistance in bovine Pasteurella multocida isolates by MALDI Biotyper antibiotic susceptibility test rapid assay (MBT-ASTRA)

Abstract

Pasteurella multocida is notorious for its role as an opportunistic pathogen in infectious bronchopneumonia, the economically most important disease facing cattle industry and leading indication for antimicrobial therapy. To rationalize antimicrobial use, avoiding imprudent use of highly and critically important antimicrobials for human medicine, availability of a rapid antimicrobial susceptibility test is crucial. The objective of the present study was to design a MALDI Biotyper antibiotic susceptibility test rapid assay (MBT-ASTRA) procedure for tetracycline resistance detection in P. multocida. This procedure was validated on 100 clinical isolates with MIC-gradient strip test, and a comparison with disk diffusion was made. Sensitivity and specificity of the MBT-ASTRA procedure were 95.7% (95% confidence interval (CI) = 89.8–101.5) and 100% (95% CI = 100–100), respectively, classifying 98% of the isolates correctly after only three hours of incubation. Sensitivity and specificity of disk diffusion were 93.5% (95% CI = 86.3–100.6) and 96.3% (95% CI = 91.3–101.3) respectively, classifying 95% of the isolates correctly. In conclusion, this MBT-ASTRA procedure has all the potential to fulfil the need for a rapid and highly accurate tetracycline susceptibility testing in P. multocida to rationalize antimicrobial use in outbreaks of bronchopneumonia in cattle or other clinical presentations across species.

Agreement of Quantitative and Qualitative Antimicrobial Susceptibility Testing Methodologies: The Case of Enrofloxacin and Avian Pathogenic Escherichia coli

“…The procedure was carried out as described previously (DeMars et al, 2016;Van Driessche et al, 2018). In brief, APEC strains were grown overnight on McConkey agar (Oxoid, Thermo Fisher Scientific, Merelbeke, Belgium).…”

“…Antimicrobial susceptibility testing (AST) is essential for rational antimicrobial drug usage and a mandatory condition to continue employing fluoroquinolones as treatment option in veterinary medicine in some countries (Royal, 2016;Van Driessche et al, 2018). Antimicrobial susceptibility testing can be performed either quantitatively or qualitatively.…”

“…Quantitative testing methods provide numerical MIC values, which are more accurate descriptors of bacterial resistance levels. At present, the agar and microbroth dilution tests are regarded as the gold standard for quantitatively determining MIC values of different bacteria (Liu et al, 2014;Lallemand et al, 2016;Clinical and Laboratory Standards Institute, 2018;Van Driessche et al, 2018;Miftahussurur et al, 2020). However, these techniques are elaborate and require specialized equipment.…”

“…However, these techniques are elaborate and require specialized equipment. The MIC-gradient strip test has gained acceptance as another quantitative method for susceptibility testing (Kelly et al, 1999), although it is not held in the same regard as the established agar and microbroth dilution methodologies (Van Driessche et al, 2018). However, there is consensus on the overall agreement between the strip test and the microbroth and agar dilution techniques for different "bug-drug" combinations (Jones et al, 1996;Hoogkamp-Korstanje et al, 1997;Kelly et al, 1999;Glupczynski et al, 2002;Liu et al, 2014;Deak et al, 2015).…”

Rationele antimicrobiële therapie voor sepsis bij runderen in het licht van de nieuwe wetgeving over kritisch belangrijke antibiotica

“…Fast diagnostics are crucial to guide rational sepsis treatment. With MBT-ASTRA, a MALDI TOF procedure, an antibiogram could be reached as fast as one to three hours post culture initiation, also for veterinary pathogens like Pasteurella multocida (Sparbier et al, 2016;Van Driessche et al, 2017). This technology completely meets the demand for fast diagnostics, which is also crucial in food animal medicine if prudent antimicrobial use needs to be combined with maximization of patient survival chances.…”