A study on the resistance mechanism of Morganella morganii to carbapenems

Abstract

Abstract: Objective To investigate resistance mechanisms to carbapenems and transmission modes of 3 Morganella morganii isolates (M1， M2 and M3) isolated from patients in surgical intensive care unit (SICU). Methods The susceptibility of antimicrobial agents was detected by agar dilution method. The molecular epidemiological characteristics of the isolates were analyzed by enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR). The encoding genes of beta-lactamases were analyzed by specific PCR amplification. The transitivity and homology of plasmids were detected by plasmid conjugation and restriction endonuclease analysis. Genetic environments around bla_KPC were analyzed by sequencing. The changes of outer membrane permeability were analyzed by the electrophoresis of outer membrane protein. Results The bla_KPC-2 was detected by PCR in all 3 isolates and their transconjugants. M1 and M2 were the same clonal type. Carbapenem-resistance was successfully transfered by conjugation experiments. The bla_KPC-2 was located on dissimilar plasmids with quite different restriction profiles. The genetic environments around bla_KPC-2 were the same. The 3 original Morganella morganii isolates showed a loss of 38 000 outer membrane proteins, and additional 36 000 outer membrane proteins appeared compared with the Morganella morganii isolates susceptible to carbapenems. Conclusions The bla_KPC-2is detected in the 3 Morganella morganii isolates belonging to 2 clonal types . This gene encoded by dissimilar plasmids is likely located on the same mobile genetic elements by which bla_KPC-2 is possibly transferred between different plasmids. The KPC-2-producing and lack of outer membrane proteins together cause these 3 Morganella morganii isolates resistant to carbapenems possibly.

Key words: Morganella morganii, Carbapenem, Beta-lactamase, Drug resistance

References

［1］ Queenan AM，Bush K. Carbapenemases: the versatile beta-lactamases［J］. Clin Microbiol Rev， 2007， 20(3):440-458.

［2］ Miriagou V，Cornaglia G，Edelstein M， et al. Acquired carbapenemases in Gram-negative bacterial patho-gens: detection and surveillance issues［J］. Clin Microbiol Infect， 2010， 16(2):112-122.

［3］ Nordmann P，Cuzon G，Naas T. The real threat of Klebsiella pneumoniae carbapenemase-producing bacteria［J］. Lancet Infect Dis， 2009， 9(4):228-236.

［4］ Wei G，Pan L，Du H， et al. ERIC-PCR fingerprinting-based community DNA hybridization to pinpoint genome-specific fragments as molecular markers to identify and track populations common to healthy human guts［J］. J Microbiol Methods， 2004， 59(1):91-108.

［5］ Smith Moland E，Hanson ND，Herrera VL， et al. Plasmid-mediated， carbapenem-hydrolysing beta-lactamase， KPC-2， in Klebsiella pneumoniae isolates［J］. J Antimicrob Chemother， 2003， 51(3):711-714.

［6］ Senda K，Arakawa Y，Ichiyama S， et al. PCR detection of metallo-beta-lactamase gene (blaIMP) in gram-negative rods resistant to broad-spectrum beta-lactams［J］. J Clin Microbiol， 1996， 34(12):2909-2913.

［7］ Tsakris A，Pournaras S，Woodford N， et al. Outbreak of infections caused by Pseudomonas aeruginosa producing VIM-1 carbapenemase in Greece［J］. J Clin Microbiol， 2000， 38(3):1290-1292.

［8］ Queenan AM， Torres-Viera C， Gold HS， et al. SME-type carbapenem-hydrolyzing class A beta-lactamases from geographically diverse Serratia marcescens strains［J］. Antimicrob Agents Chemother，2000，44(11):3035-3039.

［9］ Poirel L， Heritier C， Tolun V， et al. Emergence of oxacillinase-mediated resistance to imipenem in Klebsiella pneumoniae［J］. Antimicrob Agents Chemother，2004，48(1):15-22.

［10］ Woodford N， Tierno PM， Young K， et al. Outbreak of Klebsiella pneumoniae producing a new carbapenem hydrolyzing class A beta-lactamase，KPC-3，in a New York Medical Center［J］. Antimicrob Agents Chemother，2004，48(12):4793-4799.

［11］ Poirel L，Le Thomas I，Naas T，et al. Biochemical sequence analyses of GES-1， a novel class A extended-spectrum beta-lactamase，and the class 1 integron In52 from Klebsiella pneumoniae ［J］. Antimicrob Agents Chemother，2000，44(3):622-632.

［12］ Rasheed JK，Jay C，Metchock B， et al. Evolution of extended-spectrum beta-lactam resistance (SHV-8) in a strain of Escherichia coli during multiple episodes of bacteremia［J］. Antimicrob Agents Chemother，1997，41(3):647-653.

［13］〖JP4〗Nuesch-Inderbinen MT，Hachler H，Kayser FH. Detection of genes coding for extended-spectrum SHV beta-lactamases in clinical isolates by a molecular genetic method， and comparison with the E test［J］. Eur J Clin Microbiol Infect Dis， 1996， 15(5):398-402.

［14］ Yu Y，Ji S，Chen Y，et al.Resistance of strains producing extended-spectrum beta-lactamases and genotype distribution in China［J］. J Infect， 2007， 54(1):53-57.

［15］ Hernandez-Alles S，Alberti S，Alvarez D， et al. Porin expression in clinical isolates of Klebsiella pneumoniae ［J］. Microbiology，1999，145 (Pt 3):673-679.

［16］〖JP4〗Shen P，Wei Z，Jiang Y， et al. Novel genetic environment of the carbapenem-hydrolyzing beta-lactamase KPC-2 among Enterobacteriaceae in China［J］. Antimicrob Agents Chemother，2009，53(10):4333-4338.

［17］ Del Mar Tavio M，Vila J，Ruiz J， et al. Decreased permeability and enhanced proton-dependent active efflux in the development of resistance to quinolones in Morganella morganii ［J］. Int J Antimicrob Agents， 2000， 14(2):157-160.

［18］ Naas T，Cuzon G，Villegas MV， et al. Genetic structures at the origin of acquisition of the beta-lactamase blaKPC gene［J］. Antimicrob Agents Chemother， 2008， 52(4):1257-1263.

［19］ Munoz-Price LS，Quinn JP. The spread of Klebsiella pneumoniae carbapenemases: a tale of strains， plasmids， and transposons［J］. Clin Infect Dis， 2009， 49(11):1739-1741.

［20］〖JP4〗Mitsuyama J，Hiruma R，Yamaguchi A，et al.Identification of porins in outer membrane of Proteus，Morganella， and Providencia spp. and their role in outer membrane permeation of beta-lactams［J］. Antimicrob Agents Chemother，1987，31(3):379-384.

[1]	WANG Chao, ZHAO Yu. Research progress on formation mechanism and eradication strategy of bacterial persisters [J]. Laboratory Medicine, 2023, 38(8): 790-795.
[2]	ZHU Yurong, ZHANG Dan, HE Yaxing, LI Jingjing, HUANG Jing, LI Ting, LIU Peng, LIU Ronghua. Classification and epidemiology characteristics of Klebsiella pneumoniae isolated from clinic [J]. Laboratory Medicine, 2023, 38(5): 435-440.
[3]	FANG Yongmei, ZHANG Yan, XU Xueying, ZHONG Feng. Correlation of Staphylococcus aureus drug resistance with drug resistance genes and virulence genes [J]. Laboratory Medicine, 2023, 38(4): 357-361.
[4]	WANG Jiawei, ZHU Weinan, CHEN Yingying, JI Ping, WANG Ying. Clinical characteristics，molecular typing and drug resistance genes of carbapenem-resistant Klebsiella pneumoniae isolated from blood culturing [J]. Laboratory Medicine, 2023, 38(4): 362-367.
[5]	REN Yanfei, ZHANG Min, YANG Tao, LI Rongkai, LIANG Xin. Analysis of pathogenic epidemiology of patients with lower respiratory tract infection in respiratory intensive care unit [J]. Laboratory Medicine, 2023, 38(2): 157-162.
[6]	DENG Chen, DAI Jiaze, QIU Lihong, SHEN Weiting, CAI Mufa, LUO Wenying. Molecular epidemic characteristics of hypervirulent Klebsiella pneumoniae in Zhanjiang [J]. Laboratory Medicine, 2023, 38(11): 1026-1031.
[7]	RU Haohao, CHEN Lianyong, YANG Xing, CHEN Tao, YAN Shuangqun, XU Lin. Analysis of rifampicin resistance in Mycobacterium tuberculosis determined by Xpert MTB/RIF and phenotypic drug susceptibility test [J]. Laboratory Medicine, 2023, 38(11): 1032-1035.
[8]	WANG Su, DING Li, JIANG Wenrong, MIAO Yingxin, ZHANG Yanmei, ZHAO Hu. Research progress on drug resistance and treatment strategies of hypervirulent Klebsiella pneumoniae [J]. Laboratory Medicine, 2023, 38(1): 81-86.
[9]	HAO Jiahui, YANG Zehua. Research progress of bacterial integron determination methods [J]. Laboratory Medicine, 2022, 37(9): 887-891.
[10]	XU Qi, PAN Fen, SUN Yan, SHI Yingying, YU Fangyuan, ZHANG Hong. Methodology evaluation of screening techniques for CRE colonized in intestinal tract [J]. Laboratory Medicine, 2022, 37(8): 761-765.
[11]	LI Hua, WANG Weiliang, XIE Bei, YANG Yu, MENG Fanrong, WANG Nan, LIU Zhihui, ZHANG Yanbin. Feasibility of the determination of heterogeneous drug resistance to rifampicin in Mycobacterium tuberculosis by flow cytometry [J]. Laboratory Medicine, 2022, 37(6): 577-582.
[12]	SUN Xiaodan, LIU Yiwen, HE Yiqing, DU Yan, ZHANG Guoliang, GAO Feng, YANG Cuixia. Research progress on the role of estrogen receptor in osteosarcoma [J]. Laboratory Medicine, 2022, 37(4): 313-318.
[13]	WANG Jianying, WANG Shanshan, ZHU Jun, HUANG Xiaochun, WAN Yuxiang, LIU Yun. Distribution and drug resistance of respiratory infection pathogens in patients with lung cancer after chemotherapy [J]. Laboratory Medicine, 2022, 37(4): 356-359.
[14]	WEN Qinghui, LI Fengying, LI Yanchang, HUANG Jinyin, WANG Fengping. Drug susceptibility of multidrug-resistant ESBL-producing Escherichia coli to antimicrobial agents restored by different concentrations of EDTA [J]. Laboratory Medicine, 2022, 37(3): 205-208.
[15]	SUN Ying, LI Yi, YAN Wenjuan, JING Nan, MA Bing. Analysis of drug resistance of rare CRE and clinical application of carbapenem screening test in Henan [J]. Laboratory Medicine, 2022, 37(2): 146-149.