Molecular and Functional Characterization of a Novel Plasmid-Borne -Like Gene, , in a Clinical Strain of .
Xi Lin, Junwan Lu, Changrui Qian, Hailong Lin, Qiaoling Li, Xueya Zhang, Hongmao Liu, Zhewei Sun, Danying Zhou, Wei Lu, Mei Zhu, Hailin Zhang, Teng Xu, Kewei Li, Qiyu Bao, Li Lin
Author Information
Xi Lin: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Junwan Lu: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Changrui Qian: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Hailong Lin: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Qiaoling Li: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Xueya Zhang: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Hongmao Liu: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Zhewei Sun: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Danying Zhou: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Wei Lu: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Mei Zhu: Department of Clinical Laboratory, Zhejiang Hospital, Hangzhou, Zhejiang, 310013, People's Republic of China.
Hailin Zhang: Institute of Biomedical Informatics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Teng Xu: Institute of Translational Medicine, Baotou Central Hospital, Baotou, 014040, People's Republic of China.
Kewei Li: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Qiyu Bao: Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education of China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
Li Lin: Institute of Biomedical Informatics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People's Republic of China.
PURPOSE: An increasing frequency of antibiotic resistance has been observed in both clinical and environmental isolates in recent years. However, there are still very few in-depth studies regarding the role of plasmids in the antibiotic resistance of . Hence, we investigated the molecular and functional characterization of a multidrug-resistant plasmid encoding an NDM-like metallo-β-lactamase, AFM-1, in the clinical isolate SS332. METHODS: The minimum inhibitory concentrations (MICs) of 24 antibiotics against SS332 were measured by the agar dilution method. The genome of SS332 was sequenced with PacBio and Illumina platforms. Six plasmid-borne antimicrobial resistance genes were chosen for cloning, including , , (E), (E), , and . Phylogenetic analysis, amino acid sequence alignment, and comparative genomic analysis were performed to elucidate the active site requirements and genetic context of the gene. RESULTS: SS332 showed high levels of resistance to 15 antibiotics, especially those with MIC levels at or above 1024 μg/mL, including ampicillin, cefazolin, ceftriaxone, aztreonam, spectinomycin, and roxithromycin. Six plasmid-borne resistance genes from were verified to be functional in DH5α. AFM-1 shared 86% amino acid identity with NDM-1 and showed resistance to ampicillin, cefazolin, cefoxitin, and ceftazidime. In addition, the gene was associated with three different novel IS-like elements, designated IS, IS and ∆IS, which may be involved in the acquisition and mobilization of the gene. CONCLUSION: Our investigation showed that plasmid-borne resistance genes can contribute to antibiotic resistance in SS332. A novel -like gene, , was verified to be functional and associated with novel IS-like elements. This fact indicated the risk of spread of genes and IS-like elements.
