Evaluation of Commercial Products for Colistin and Polymyxin B Susceptibility Testing for mcr-Positive and Negative Escherichia coli and Klebsiella pneumoniae in China.

Purpose: To evaluate the performance of five widespread commercial products for colistin and polymyxin B susceptibility testing in China for mcr-positive and -negative Escherichia coli and Klebsiella pneumoniae. Methods: A total of 132 E. coli and 83 K. pneumoniae strains (including 68 mcr-1-positive E. coli and 28 mcr-8-positive K. pneumoniae) were collected. We analysed the performance of colistin susceptibility (with Vitek 2 and Phoenix M50) and the performance of polymyxin B susceptibility (with DL-96II, MA120, and a Polymyxin B Susceptibility Test strip; POL E-strip). Broth microdilution was used as the gold standard. Categorical agreement (CA), essential agreement (EA), major error (ME), and very major error (VME) were calculated for comparisons. Results: For E. coli, the total CA, EA, ME, and VME to colistin were as follows: Vitek 2, 98.5%/98.5%/0%/2.9%; and Phoenix M50, 98.5%/97.7%/0%/2.9%. The total CA, EA, ME, and VME to polymyxin B were as follows: POL E-strip, 99.2%/63.6%/1.6%/0%; MA120, 70.0%/-/0%/58.8%; and DL-96II, 80.2%/-/1.6%/36.8%. Only Vitek 2 and Phoenix M50 presented satisfactory performances for mcr-1-positive E. coli. For K. pneumoniae, the total CA, EA, ME, and VME to colistin were as follows: Vitek 2, 73.2%/72.0%/0%/61.6%; and Phoenix M50, 74.7%/74.7%/0%/58.3%. The total CA, EA, ME, and VME to polymyxin B were as follows: POL E-strip, 91.6%/74.7%/2.1%/16.7%; MA120, 92.8%/-/2.1%/13.9%; and DL-96II, 92.2%/-/2.1%/8.3%. All systems were unsatisfactory for mcr-8-positive K. pneumoniae. When the susceptibility of mcr-negative strains was tested, all systems presented excellent performance. Conclusion: Vitek 2 and Phoenix M50 with colistin for E. coli showed acceptable performance regardless of mcr-1 expression, while DL-96II, MA120, and the POL E-strip performed worse for mcr-1-positive strains. Furthermore, mcr-8 greatly affected the performance of all systems with both colistin and polymyxin B for K. pneumoniae isolates.

Epidemiology and genotypic characteristics of carbapenem resistant Enterobacterales in Henan, China: a multicentre study.

OBJECTIVES: To identify the carbapenemase and colistin resistance genes and epidemiological status of carbapenem-resistant Enterobacterales (CRE) among 7 secondary and 11 tertiary hospitals in Henan province, China. METHODS: CRE isolates and clinical data of infected patients were collected from 7 secondary and 11 tertiary hospitals in Henan from July to September 2019 and analysed retrospectively. Polymerase chain reaction (PCR) and gene sequencing were performed to detect carbapenemase and colistin resistance genes; multilocus sequence typing was also performed. RESULTS: In total, 238 nonduplicate CRE isolates were collected mainly from the respiratory tract (54.20%) and blood (18.91%) of CRE-infected patients, half of them aged >65 years (45.80%). Carbapenem-resistant Klebsiella pneumoniae (CRKP) was the most common CRE (184 isolates, 77.31%) with constituent ratios of 84.38% and 72.54% in secondary and tertiary hospitals, respectively. In 184 CRKP isolates, blaKPC-2 (89.13%) was the dominant carbapenemase gene, and ST11 (71.74%) was the most prevalent sequence type (ST), with constituent ratios of 83.95% and 62.14% in secondary and tertiary hospitals, respectively. In 29 carbapenem-resistant Escherichia coli (CREC) isolates, blaNDM-5 (58.62%) and ST2 (31.03%) were prevalent. Four CRKP isolates and one CREC isolate were colistin-resistant and carried the plasmid-mediated mcr-1 gene. CONCLUSION: Our results showed a wide spread of CRKP-ST11 with KPC-2 carbapenemase in the analysed 18 hospitals. The CRKP constituent ratio, CRKP-STs, and CREC carbapenemase genes between secondary and tertiary hospitals showed significant differences. The emergence of a colistin-resistant CRKP with plasmid-mediated resistance gene mcr-1 is of serious concern due to the limited treatment options.

Molecular Mechanism of Polymyxin Resistance in Multidrug-Resistant Klebsiella pneumoniae and Escherichia coli Isolates from Henan Province, China: A Multicenter Study.

PURPOSE: To evaluate polymyxin-resistant Klebsiella pneumoniae and Escherichia coli prevalence and characteristics in the Henan province, China. MATERIALS AND METHODS: A total of 2301 bacterial isolates collected at six hospitals were assessed. Their response to polymyxin was evaluated by minimum inhibitory concentration (MIC) analysis, and the mobilized colistin resistance (mcr) and carbapenemase gene were explored. Mutations on mgrB, phoPQ, pmrAB, and crrAB in polymyxin-resistant K. pneumoniae were detected by PCR. phoP, phoQ, pmrK, pmrA, pmrB, and pmrC transcriptional levels were quantified by RT-qPCR. Pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing were performed to determine the phylogenetic relationship between the polymyxin-resistant isolates. RESULTS: Of the E. coli and K. pneumoniae isolates identified, 0.3% and 1.4% were polymyxin-resistant, respectively, with MICs of 4-64 µg/mL. All polymyxin-resistant isolates were susceptible to tigecycline. Four E. coli isolates were mcr-1-positive and one was carbapenem-resistant, carrying bla NDM-5 and mcr-1. One K. pneumoniae isolate was mcr-1-positive and nine were carbapenem-resistant (PRCRKP), carrying bla KPC-2 but not mcr-1. The five E. coli isolates belonged to four sequence types (ST2, ST132, ST632, and ST983). All PRCRKP isolates belonged to ST11. However, all 16 isolates belonged to different PFGE types with <95% genetic similarity. Insertion sequences in mgrB were detected in nine (81.8%) polymyxin-resistant K. pneumoniae samples. Colistin resistance was linked with pmrHFIJKLM operon upregulation, with phoP, phoQ, and pmrK being overexpressed in all but one of the polymyxin-resistant K. pneumoniae samples. Furthermore, 33.3% of patients carrying polymyxin-resistant isolates had previously used polymyxin, and 66.7% patients displayed good clinical outcomes. CONCLUSION: The K. pneumoniae polymyxin resistance rate was slightly higher than that of E. coli and mcr-1 was more common in E. coli than in K. pneumoniae. Moreover, the insertion of ISkpn14 into mgrB may be the main contributor to polymyxin-resistance in K. pneumoniae in Henan.