Carbapenem-resistance mechanisms and molecular typing of Escherichia coli producing New Delhi metal β-lactamase
FU Fen-rui1, 2, ZHANG Ya1, 3, PAN Yu-hong1, 4, CAO Ying-ping1, 4, ZHENG Pei-zheng1, 4
1. Fujian Medical University Union Hospital, Fuzhou 350001, China; 2. Ningde Municipal Hospital, Ningde 352000, China; 3. Jiujiang University Affiliated Hospital, Jiujiang 332000, China; 4. School of Medical Technology and Engineering, Fujian Medical University, Fuzhou 350004,China
Abstract:The aim of this study was to investigate the resistance mechanisms and molecular epidemiological features of Escherichia coli strains producing New Delhi metal β-lactamase (NDM) collected in a third class A hospital. NDM genes were screened with PCR in carbapenem-resistant Escherichia coli strains. Types of NDM genes were identified by sequencing. Other carbapenem-resistance related genes and extended-spectrum-β-lactamase genes were detected by PCR for NDM gene positive strains. An expression system was used to evaluate the imipenem-hydrolysis ability of NDM-1, NDM-6 and NDM-9. Molecular typing of Escherichia coli producing NDM was performed with the multilocus sequence typing (MLST) method. Eight strains of Escherichia coli producing NDM were identified, among which six were NDM-6 strains, and the remainder were NDM-9 strains. Four strains of Escherichia coli were found to simultaneously carry NDM-6, TEM and CTX-M-1 genes, and one strain was found to simultaneously carry NDM-6, TEM and CTX-M-9 genes. Recombinant Escherichia coli strains expressing NDM-1, NDM-6 and NDM-9 genes were resistant to imipenem, with minimum inhibitory concentrations of 32 μg/mL, 128 μg/mL and 64 μg/mL, respectively. MLST revealed four distinct sequence types (STs), among which five strains of ST101 were collected from the neurological unit, and the remaining three strains were ST226, ST648 and ST1284, respectively. This study reports the first identification of Escherichia coli simultaneously carrying NDM-6, TEM and CTX-M-1 genes, or NDM-6, TEM and CTX-M-9 genes. Notably, NDM-6 was associated with the highest imipenem hydrolysis ability, and was followed by NDM-9 and NDM-1. The MLST results indicated that a nosocomial infection of carbapenem-resistant Escherichia coli occurred locally in a third class A hospital.
傅芬蕊, 张娅, 潘玉红, 曹颖平, 郑培烝. 产新德里金属β-内酰胺酶大肠埃希菌耐碳青霉烯类药物相关机制研究和分子分型[J]. 中国人兽共患病学报, 2021, 37(1): 53-59.
FU Fen-rui, ZHANG Ya, PAN Yu-hong, CAO Ying-ping, ZHENG Pei-zheng. Carbapenem-resistance mechanisms and molecular typing of Escherichia coli producing New Delhi metal β-lactamase. Chinese Journal of Zoonoses, 2021, 37(1): 53-59.
[1] Tan K, Nguyen J, Nguyen K, et al.Prevalence of the carbapenem-heteroresistant phenotype among ESBL-producing Escherichia coli and Klebsiella pneumoniae clinical isolates[J]. J Antimicrob Chemother, 2020,75(6):1506-1512. DOI:10.1093/jac/dkaa048 [2] Li J, Yu T, Tao XY, et al.Emergence of an NDM-5-producing Escherichia coli sequence type 410 clone in infants in a children’s hospital in China[J]. Infect Drug Resist, 2020,13:703-710. DOI:10.2147/IDR.S244874 [3] Woodford N, Ellington MJ, Coelho JM, et al.Multiplex PCR for genes encoding prevalent OXA carbapenemases in Acinetobacter spp[J]. Int J Antimicrob Agents, 2006,27(4):351-353. DOI:10.1016/j.ijantimicag.2006.01.004 [4] Brown S, Young HK, Amyes SG.Characterisation of OXA-51, a novel class D carbapenemase found in genetically unrelated clinical strains of Acinetobacter baumannii from Argentina[J]. Clin Microbiol Infect, 2005,11(1):15-23. DOI:10.1111/j.1469-0691.2004.01016.x [5] Higgins PG, Lehmann M, Seifert H.Inclusion of OXA-143 primers in a multiplex polymerase chain reaction (PCR) for genes encoding prevalent OXA carbapenemases in Acinetobacter spp[J]. Int J Antimicrob Agents, 2010,35(3):305. DOI:10.1016/j.ijantimicag.2009.10.014 [6] Mendes RE, Kiyota KA, Monteiro J, et al.Rapid detection and identification of metallo-beta-lactamase-encoding genes by multiplex real-time PCR assay and melt curve analysis[J]. J Clin Microbiol, 2007,45(2):544-547. DOI:10.1128/JCM.01728-06 [7] Hornsey M, Phee L, Wareham DW.A novel variant, NDM-5, of the New Delhi metallo-β-lactamase in a multidrug-resistant Escherichia coli ST648 isolate recovered from a patient in the United Kingdom[J]. Antimicrob Agents Chemother, 2011,55(12):5952-5954. DOI:10.1128/AAC.05108-11 [8] Kaase M, Nordmann P, Wichelhaus TA, et al.NDM-2 carbapenemase in Acinetobacter baumannii from Egypt[J]. J Antimicrob Chemother, 2011,66(6):1260-1262. DOI:10.1093/jac/dkr135 [9] Bhattacharjee A, Sen MR, Prakash P, et al.Role of beta-lactamase inhibitors in enterobacterial isolates producing extended-spectrum beta-lactamases[J]. J Antimicrob Chemother, 2008,61(2):309-314. DOI:10.1093/jac/dkm494 [10] Tonki M, Mohar B, Šiško-Kraljevi K, et al.High prevalence and molecular characterization of extended-spectrum β-lactamase-producing Proteus mirabilis strains in southern Croatia[J]. J Med Microbiol, 2010,59(Pt 10):1185-1190. DOI:10.1099/jmm.0.016964-0 [11] Park YJ, Lee S, Kim YR, et al.Occurrence of extended-spectrum (beta)-lactamases and plasmid-mediated AmpC (beta)-lactamases among Korean isolates of Proteus mirabilis[J]. J Antimicrob Chemother, 2006,57(1):156-158. DOI:10.1093/jac/dki408 [12] Garrec H, Drieux-Rouzet L, Golmard JL, et al.Comparison of nine phenotypic methods for detection of extended-spectrum beta-lactamase production by Enterobacteriaceae[J]. J Clin Microbiol, 2011,49(3):1048-1057. DOI:10.1128/JCM.02130-10 [13] Navon-Venezia S, Chmelnitsky I, Leavitt A, et al.Dissemination of the CTX-M-25 family beta-lactamases among Klebsiella pneumoniae, Escherichia coli and Enterobacter cloacae and identification of the novel enzyme CTX-M-41 in Proteus mirabilis in Israel[J]. J Antimicrob Chemother, 2008,62(2):289-295. DOI:10.1093/jac/dkn182 [14] Goren MG, Navon-Venezia S, Chmelnitsky I, et al.Carbapenem-resistant KPC-2-producing Escherichia coli in a Tel Aviv Medical Center, 2005 to 2008[J]. Antimicrob Agents Chemother, 2010,54(6):2687-2691. DOI:10.1128/AAC.01359-09 [15] Zou H, Jia X, Liu H, et al.Emergence of NDM-5-producing Escherichia coli in a teaching hospital in Chongqing, China: IncF-type plasmids may contribute to the prevalence of blaNDM-5[J]. Front Microbiol, 2020,11:334. DOI:10.3389/fmicb.2020.00334 [16] Williamson DA, Sidjabat HE, Freeman JT, et al.Identification and molecular characterisation of New Delhi metallo-β-lactamase-1 (NDM-1)-and NDM-6-producing Enterobacteriaceae from New Zealand hospitals[J]. Int J Antimicrob Agents, 2012,39(6):529-533. DOI:10.1016/j.ijantimicag.2012.02.017 [17] Wang X, Li H, Zhao C, et al.Novel NDM-9 metallo-β-lactamase identified from a ST107 Klebsiella pneumoniae strain isolated in China[J]. Int J Antimicrob Agents, 2014,44:90-91. DOI:10.1016/j.ijantimicag.2014.04.010 [18] Lin YC, Kuroda M, Suzuki S, et al.Emergence of an Escherichia coli strain co-harbouring mcr-1 and blaNDM-9 from a urinary tract infection in Taiwan[J]. J Glob Antimicrob Resist, 2019,16:286-290. DOI:10.1016/j.jgar.2018.10.003 [19] Qamar MU, Walsh TR, Toleman MA, et al.Dissemination of genetically diverse NDM-1,-5,-7 producing-Gram-negative pathogens isolated from pediatric patients in Pakistan[J]. Future Microbiol, 2019,14(1):691-704. DOI:10.2217/fmb-2019-0012 [20] Peirano G, Pitout JDD.Extended-Spectrum β-Lactamase-Producing Enterobacteriaceae: update on molecular epidemiology and treatment options[J]. Drugs, 2019,79(14):1529-1541. DOI:10.1007/s40265-019-01180-3 [21] Mizuno Y, Yamaguchi T, Matsumoto T.A first case of New Delhi metallo-β-lactamase-7 in an Escherichia coli ST648 isolate in Japan[J]. J Infect Chemother, 2014,20(12):814-816. DOI:10.1016/j.jiac.2014.08.009