Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, P.R. China, Key Laboratory of Livestock and Poultry Multi-omics of MARA.
Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, P.R. China, Key Laboratory of Livestock and Poultry Multi-omics of MARA; Department of Genetic and Genomes Biology, University of Leicester.
J Vis Exp. 2023 Jul 21(197). doi: 10.3791/64785.
To improve the efficiency of antimicrobial susceptibility testing (AST) and phage high-throughput screening for resistant bacteria and to reduce the detection cost, an intelligent high-throughput AST/phage screening system, including a 96-dot matrix inoculator, image acquisition converter, and corresponding software, was developed according to AST criteria and the breakpoints of resistance (R) formulated by the Clinical & Laboratory Standards Institute (CLSI). AST and statistics of minimum inhibitory concentration (MIC) distributions (from R/8 to 8R) of 1,500 Salmonella strains isolated from poultry in Shandong, China, against 10 antimicrobial agents were carried out by the intelligent high-throughput AST/phage screening system. The Lar index, meaning "less antibiosis, less resistance and residual until little antibiosis", was obtained by calculating the weighted average of each MIC and dividing by R. This approach improves accuracy in comparison with using the prevalence of resistance to characterize the antimicrobial resistance (AMR) degree of highly resistant strains. For the strains of Salmonella with high AMR, lytic phages were efficiently screened from the phage library by this system, and the lysis spectrum was computed and analyzed. The results showed that the intelligent high-throughput AST/phage screening system was operable, accurate, highly efficient, inexpensive, and easy to maintain. Combined with the Shandong veterinary antimicrobial resistance monitoring system, the system was suitable for scientific research and clinical detection related to AMR.
为提高抗菌药物敏感性试验（AST）和噬菌体高通量筛选耐药菌的效率，降低检测成本，根据 AST 标准和临床实验室标准化研究所（CLSI）制定的耐药（R）临界点，我们开发了一种智能高通量 AST/噬菌体筛选系统，包括 96 孔矩阵接种器、图像采集转换器和相应的软件。利用智能高通量 AST/噬菌体筛选系统，对中国山东家禽分离的 1500 株沙门氏菌对 10 种抗菌药物的最小抑菌浓度（MIC）分布（从 R/8 到 8R）进行了 AST 和统计。Lar 指数是通过计算每个 MIC 的加权平均值并除以 R 得到的，其含义是“较少的抗生素，较少的耐药性和残留抗生素直至耐药性较低”。与使用耐药率来描述高度耐药菌株的抗菌药物耐药性（AMR）程度相比，该方法提高了准确性。对于 AMR 水平较高的沙门氏菌菌株，该系统能够从噬菌体文库中有效筛选出溶菌噬菌体，并计算和分析其裂解谱。结果表明，智能高通量 AST/噬菌体筛选系统操作方便、准确高效、价格低廉、易于维护。结合山东兽医抗菌药物耐药性监测系统，该系统适用于与 AMR 相关的科研和临床检测。
