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The determination of gyrA and parC mutations and the prevalence of plasmid-mediated quinolone resistance genes in carbapenem resistant Klebsiella pneumonia ST11 and ST76 strains isolated from patients in Heilongjiang Province, China.
Infect Genet Evol. 2020 08; 82:104319.IG

Abstract

BACKGROUND

There is increasing resistance to carbapenems among Klebsiella pneumoniae,and fluoroquinolones (FQ) are increasingly used to treat infections from extended-spectrum β- lactamase(ESBLs) and carbapenemase-producing Klebsiella pneumoniae. However, the acquisition of plasmid-mediated quinolone resistance (PMQR) or the spontaneous mutation of the quinolone resistance-determining regions (QRDR) of the gyrA and parC genes can severely affect the therapeutic effect of quinolones. The goal of this study was to investigate the molecular determinants of FQ resistance(FQ-R) in carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates from Heilongjiang Province,China.

MATERIALS AND METHODS

We isolated 40 strains of CRKP from a treatment center in the eastern part of Heilongjiang Province from January 2016 to December 2018. The VITEK2 Compact analyzer was used to identify and detect drug sensitivity. Different types of drug resistance genes were detected by polymerase chain reaction (PCR). PCR and DNA sequencing were used to assess the presence of qnrA, qnrB, qnrS,qepA and acc(6') Ib-cr genes,which are plasmid-encode genes that can contribute to resistance. The sequences of gyrA and parC genes were sequenced and compared with the sequences of standard strains to determine if mutations were present.Multi-site sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were performed on the strains to assess homology.

RESULTS

The isolated CRKP strains showed rates of resistance to fluoroquinolones of 22.5% to 42.5%. The resistance rate of ciprofloxacin was significantly higher than that of levofloxacin.Nine CRKP strains (22.5%) showed co-resistance to ciprofloxacin and levofloxacin.The quinolone resistant strains were screened for plasmid-encoded genes that can contribute to resistance (PMQR genes).Among the 17 quinolone resistant strains,one strain contained no PMQR genes,twelve strains contained two PMQR genes,and four strains contained four PMQR genes.Acc (6') Ib-cr was the most frequently detected PMQR gene, detected in 95% of strains tested (38 of 40) and in 94.1% of the quinolone-resistant strains (16 of 17). The qepA gene encoding an efflux pump was not detected in any strains.No isolate carried five different PMQRs simultaneously.Changes of S83I and D87G changes in gyrA, and the S80I change in parC,which were mediated by QRDR,were identified in two isolates,which showed resistance to both ciprofloxacin and levofloxacin.Most of the FQ-R strains(58.8%,10/17) belong to ST(sequence type) 76, which is dominant in the local area, while all the mutant strains (100%,2/2),that differ in at least one site from standard bacteria, belong to the ST11 group. The strains were isolated from a hospital where there had been a recent outbreak of ST76 type CRKP in the neurosurgery ward and intensive care unit.

CONCLUSION

CRKP strains were identified that were insensitive or even resistant to quinolones,and this resistance is common in Heilongjiang Province of eastern China;fluoroquinolone-resistance in these clinical CRKP strains is a complex interplay between PMQR determinants and mutations in gyrA and parC.The resistance level caused by QRDR mutation is higher than that caused by PMQR, however, the high frequency of PMQR genes in the isolated CRKP strains suggests the potential for impact of these genes.PMQR determinants are often found in carbapenemase-producing or ESBLs-producing Klebsiella pneumoniae,and some resistance genes,such as:SHV,TEM, CTX-M-15,and OXA-1 are closely associated with FQ-R. Finally, geographical factors can affect the emergence and spread of PMQR and QRDR.Some genetic lineages have higher potential risks, and continuous close monitoring is required.

Authors+Show Affiliations

Department of Microbiology, The First Affiliated Hospital of Jiamusi University, Jiamusi 154002, China.Department of Microbiology, Yongchuan Hospital of Chongqing Medical University, Chongqing 402106, China.Department of Pathogenic Biology,Jiamusi University School of Basic Medicine, Jiamusi 154007, China.Department of Microbiology, The First Affiliated Hospital of Jiamusi University, Jiamusi 154002, China.Department of Microbiology, The First Affiliated Hospital of Jiamusi University, Jiamusi 154002, China.Department of Microbiology, The First Affiliated Hospital of Jiamusi University, Jiamusi 154002, China.Department of Scientific Research Section, Jiamusi University School of Clinical Medicine, Jiamusi 154007, China.Department of Microbiology, The First Affiliated Hospital of Jiamusi University, Jiamusi 154002, China.Department of Microbiology, Yongchuan Hospital of Chongqing Medical University, Chongqing 402106, China; Department of Microbiology, The First Affiliated Hospital of Jiamusi University, Jiamusi 154002, China. Electronic address: jmszxl123@163.com.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32278145

Citation

Zeng, Lingyi, et al. "The Determination of gyrA and parC Mutations and the Prevalence of Plasmid-mediated Quinolone Resistance Genes in Carbapenem Resistant Klebsiella Pneumonia ST11 and ST76 Strains Isolated From Patients in Heilongjiang Province, China." Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases, vol. 82, 2020, p. 104319.
Zeng L, Zhang J, Li C, et al. The determination of gyrA and parC mutations and the prevalence of plasmid-mediated quinolone resistance genes in carbapenem resistant Klebsiella pneumonia ST11 and ST76 strains isolated from patients in Heilongjiang Province, China. Infect Genet Evol. 2020;82:104319.
Zeng, L., Zhang, J., Li, C., Fu, Y., Zhao, Y., Wang, Y., Zhao, J., Guo, Y., & Zhang, X. (2020). The determination of gyrA and parC mutations and the prevalence of plasmid-mediated quinolone resistance genes in carbapenem resistant Klebsiella pneumonia ST11 and ST76 strains isolated from patients in Heilongjiang Province, China. Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases, 82, 104319. https://doi.org/10.1016/j.meegid.2020.104319
Zeng L, et al. The Determination of gyrA and parC Mutations and the Prevalence of Plasmid-mediated Quinolone Resistance Genes in Carbapenem Resistant Klebsiella Pneumonia ST11 and ST76 Strains Isolated From Patients in Heilongjiang Province, China. Infect Genet Evol. 2020;82:104319. PubMed PMID: 32278145.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The determination of gyrA and parC mutations and the prevalence of plasmid-mediated quinolone resistance genes in carbapenem resistant Klebsiella pneumonia ST11 and ST76 strains isolated from patients in Heilongjiang Province, China. AU - Zeng,Lingyi, AU - Zhang,Jisheng, AU - Li,Chunjiang, AU - Fu,Yanjun, AU - Zhao,Yongxin, AU - Wang,Yong, AU - Zhao,Jing, AU - Guo,Yuhang, AU - Zhang,Xiaoli, Y1 - 2020/04/08/ PY - 2020/01/14/received PY - 2020/03/17/revised PY - 2020/04/07/accepted PY - 2020/4/12/pubmed PY - 2021/10/7/medline PY - 2020/4/12/entrez KW - Carbapenem-resistance Klebsiella pneumoniae KW - MLST KW - PMQR KW - QRDR KW - Quinolone-resistance SP - 104319 EP - 104319 JF - Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases JO - Infect Genet Evol VL - 82 N2 - BACKGROUND: There is increasing resistance to carbapenems among Klebsiella pneumoniae,and fluoroquinolones (FQ) are increasingly used to treat infections from extended-spectrum β- lactamase(ESBLs) and carbapenemase-producing Klebsiella pneumoniae. However, the acquisition of plasmid-mediated quinolone resistance (PMQR) or the spontaneous mutation of the quinolone resistance-determining regions (QRDR) of the gyrA and parC genes can severely affect the therapeutic effect of quinolones. The goal of this study was to investigate the molecular determinants of FQ resistance(FQ-R) in carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates from Heilongjiang Province,China. MATERIALS AND METHODS: We isolated 40 strains of CRKP from a treatment center in the eastern part of Heilongjiang Province from January 2016 to December 2018. The VITEK2 Compact analyzer was used to identify and detect drug sensitivity. Different types of drug resistance genes were detected by polymerase chain reaction (PCR). PCR and DNA sequencing were used to assess the presence of qnrA, qnrB, qnrS,qepA and acc(6') Ib-cr genes,which are plasmid-encode genes that can contribute to resistance. The sequences of gyrA and parC genes were sequenced and compared with the sequences of standard strains to determine if mutations were present.Multi-site sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were performed on the strains to assess homology. RESULTS: The isolated CRKP strains showed rates of resistance to fluoroquinolones of 22.5% to 42.5%. The resistance rate of ciprofloxacin was significantly higher than that of levofloxacin.Nine CRKP strains (22.5%) showed co-resistance to ciprofloxacin and levofloxacin.The quinolone resistant strains were screened for plasmid-encoded genes that can contribute to resistance (PMQR genes).Among the 17 quinolone resistant strains,one strain contained no PMQR genes,twelve strains contained two PMQR genes,and four strains contained four PMQR genes.Acc (6') Ib-cr was the most frequently detected PMQR gene, detected in 95% of strains tested (38 of 40) and in 94.1% of the quinolone-resistant strains (16 of 17). The qepA gene encoding an efflux pump was not detected in any strains.No isolate carried five different PMQRs simultaneously.Changes of S83I and D87G changes in gyrA, and the S80I change in parC,which were mediated by QRDR,were identified in two isolates,which showed resistance to both ciprofloxacin and levofloxacin.Most of the FQ-R strains(58.8%,10/17) belong to ST(sequence type) 76, which is dominant in the local area, while all the mutant strains (100%,2/2),that differ in at least one site from standard bacteria, belong to the ST11 group. The strains were isolated from a hospital where there had been a recent outbreak of ST76 type CRKP in the neurosurgery ward and intensive care unit. CONCLUSION: CRKP strains were identified that were insensitive or even resistant to quinolones,and this resistance is common in Heilongjiang Province of eastern China;fluoroquinolone-resistance in these clinical CRKP strains is a complex interplay between PMQR determinants and mutations in gyrA and parC.The resistance level caused by QRDR mutation is higher than that caused by PMQR, however, the high frequency of PMQR genes in the isolated CRKP strains suggests the potential for impact of these genes.PMQR determinants are often found in carbapenemase-producing or ESBLs-producing Klebsiella pneumoniae,and some resistance genes,such as:SHV,TEM, CTX-M-15,and OXA-1 are closely associated with FQ-R. Finally, geographical factors can affect the emergence and spread of PMQR and QRDR.Some genetic lineages have higher potential risks, and continuous close monitoring is required. SN - 1567-7257 UR - https://www.unboundmedicine.com/medline/citation/32278145/The_determination_of_gyrA_and_parC_mutations_and_the_prevalence_of_plasmid_mediated_quinolone_resistance_genes_in_carbapenem_resistant_Klebsiella_pneumonia_ST11_and_ST76_strains_isolated_from_patients_in_Heilongjiang_Province_China_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1567-1348(20)30150-7 DB - PRIME DP - Unbound Medicine ER -