Concordance of disk diffusion, broth microdilution, and whole-genome sequencing for determination of in vitro antimicrobial susceptibility of Mannheimia haemolytica

纸片扩散法、肉汤微量稀释法和全基因组测序法测定溶血性曼氏杆菌体外抗菌药物敏感性的一致性

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Abstract

BACKGROUND: Extensive drug resistance (XDR) is an emerging concern with Mannheimia haemolytica, and a variety of testing methods are available for characterizing in vitro antimicrobial susceptibility. OBJECTIVES: To compare the concordance among disk diffusion, broth microdilution, and whole genome sequencing (WGS) for susceptibility testing of M. haemolytica before and after mass treatment using tulathromycin. ANIMALS: Forty-eight M. haemolytica isolates collected from high-risk beef stocker calves before and after mass treatment (metaphylaxis) using tulathromycin (Draxxin, Zoetis, Parsippany, NJ) given at the label dosage of 2.5 mg/kg body weight SC in the neck. METHODS: In vitro antimicrobial susceptibility was determined for all 48 isolates using disk diffusion, broth microdilution, and WGS. Concordance was calculated between pairs of susceptibility testing methods as follows: number of isolates classified identically by the 2 testing methods for each timepoint, divided by the number of isolates tested at that timepoint. Discordance was calculated as follows: number of isolates classified differently by the 2 testing methods for each timepoint, divided by the number of isolates tested at that timepoint. RESULTS: Concordance between testing methods ranged from 42.3% to 100%, depending on antimicrobial evaluated, timing of sample collection, and testing method used. Very major errors were identified in up to 7.7% of classifications whereas minor errors were seen in up to 50% of classifications depending on antimicrobial evaluated, timing of sample collection, and testing method used. CONCLUSIONS AND CLINICAL IMPORTANCE: Our results show that discrepancies in the results of different susceptibility testing methods occur and suggest a need for greater harmonization of susceptibility testing methods.

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