Assessment of two selective agar media to isolate colistin-resistant bovine Escherichia coli: Correlation with minimal inhibitory concentration and presence of mcr genes

Thiry, Damien; Berrah, A.; Evrard, Julien; Duprez, Jean-Noël; Mainil, Jacques; Saulmont, Marc

doi:10.1016/j.mimet.2019.03.004

Cited by 15 publications

(15 citation statements)

References 22 publications

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“…A recent study by Thiry et al . (2019) evaluated CHROMagar COL- APSE against CHROMID Colistin_R and agar disk diffusion and concluded that growth on the media corelates with resistant results on the disk diffusion and not with the presence of the mcr genes 10 . Our study supports this as growth on the CHROMagar in this study was observed for most isolates that did have any mcr genes in the PCR assay; however, this study had only one mcr- positive isolate, limiting a comprehensive comparison and conclusion.…”

Section: Discussionmentioning

confidence: 99%

Comparative Evaluation of CHROMagar COL-APSE, MicroScan Walkaway, ComASP Colistin, and Colistin MAC Test in Detecting Colistin-resistant Gram-Negative Bacteria

Sekyere

Sephofane

Mbelle

2019

Preprint

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Colistin has become a critical antibiotic for fatal Gram-negative infections owing to the proliferation of multidrug-resistant carbapenemase-producing bacteria. Thus, cheaper, faster, efficient and easier-to-use colistin diagnostics are required for clinical surveillance, diagnoses and therapeutics. The sensitivity, specificity, major error (ME), very major error (VME), categorial agreement, essential agreement, turnaround time (TAT), average cost, and required skill for four colistin resistance diagnostics viz., CHROMagar COL-APSE, ComASP Colistin, MicroScan, and Colistin MAC Test (CMT) were evaluated against broth microdilution (BMD) using 84 Gram-negative bacterial isolates. A multiplex PCR (M-PCR) was used to screen all isolates to detect the presence of the mcr-1 to mcr-5 genes. A 15-point grading scale was used to grade the tests under skill, ease, processing time etc. mcr-1 was detected by both M-PCR and CMT in a single E. coli isolate, with other PCR amplicons suggestive of mcr-2, -3 and -4 genes being also observed on the gel. The sensitivity and specificity of CHROMagar COL-APSE, MicroScan, and ComASP Colistin, were 82.05% and 66.67%, 92.31% and 76.92%, and 100% and 88.89% respectively. The MicroScan was the most expensive at a cost (per sampe tested) of R221.6 ($15.0), followed by CHROMagar COL-APSE (R118.3; $8.0), M-PCR (R75.1; $5.1), CMT (R20.1; $1.4) and ComASP Colistin (R2.64; $0.2). CHROMagar was the easiest to perform, followed by ComASP Colistin, M-PCR, MicroScan, CMT and BMD whilst M-PCR and MicroScan required higher skill. The ComASP Colistin was the best performing diagnostic test, with low VME and ME, making it recommendable for routine colistin sensitivity testing in clinical laboratories; particularly, in poorer settings. It is however limited by a TAT of 18-24 hours.

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Section: Discussionmentioning

confidence: 99%

Comparative Evaluation of CHROMagar COL-APSE, MicroScan Walkaway, ComASP Colistin, and Colistin MAC Test in Detecting Colistin-resistant Gram-Negative Bacteria

Sekyere

Sephofane

Mbelle

2019

Preprint

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“…This discrepancy could be due to the other study's use of serial www.nature.com/scientificreports www.nature.com/scientificreports/ dilutions in broth whilst we evaluated the performance of the media using cultured bacteria; this might be a limitation in our study that can be further investigated. A recent study by Thiry et al (2019) evaluated CHROMagar COL-APSE against CHROMID Colistin_R and agar disk diffusion and concluded that growth on the media corelates with resistant results on the disk diffusion and not with the presence of the mcr genes 10 . Our study supports this as growth on the CHROMagar in this study was observed for most isolates that did have any mcr genes in the PCR assay; however, this study had only one mcr-positive isolate, limiting a comprehensive comparison and conclusion.…”

Section: Isolatementioning

confidence: 99%

Comparative Evaluation of CHROMagar COL-APSE, MicroScan Walkaway, ComASP Colistin, and Colistin MAC Test in Detecting Colistin-resistant Gram-Negative Bacteria

Sekyere

Sephofane

Mbelle

2020

Sci Rep

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colistin has become a critical antibiotic for fatal Gram-negative infections owing to the proliferation of multidrug-resistant carbapenemase-producing bacteria. Thus, cheaper, faster, efficient and easier-touse colistin diagnostics are required for clinical surveillance, diagnoses and therapeutics. the sensitivity, specificity, major error (ME), very major error (VME), categorial agreement, essential agreement, turnaround time (TAT), average cost, and required skill for four colistin resistance diagnostics viz., cHRoMagar coL-APSE, ComASP Colistin, MicroScan, and Colistin MAC Test (CMT) were evaluated against broth microdilution (BMD) using 84 Gram-negative bacterial isolates. A multiplex PCR (M-PCR) was used to screen all isolates to detect the presence of the mcr-1 to mcr-5 genes. A 15-point grading scale was used to grade the tests under skill, ease, processing time etc. mcr-1 was detected by both M-pcR and cMt in a single E. coli isolate, with other pcR amplicons suggestive of mcr-2,-3 and-4 genes being also observed on the gel. The sensitivity and specificity of CHROMagar COL-APSE, MicroScan, and ComASP Colistin, were 82.05% and 66.67%, 92.31% and 76.92%, and 100% and 88.89% respectively. The MicroScan was the most expensive at a cost (per sampe tested) of R221.6 ($15.0), followed by cHRoMagar coL-APSE (R118.3; $8.0), M-PCR (R75.1; $5.1), CMT (R20.1; $1.4) and ComASP Colistin (R2.64; $0.2). CHROMagar was the easiest to perform, followed by ComASP Colistin, M-PCR, MicroScan, cMt and BMD whilst M-pcR and MicroScan required higher skill. the comASp colistin was the best performing diagnostic test, with low VME and ME, making it recommendable for routine colistin sensitivity testing in clinical laboratories; particularly, in poorer settings. It is however limited by a TAT of 18-24 hours. Subsequent to the exponential dissemination of extended-spectrum β-lactamases (ESBLs) in Gram-negative bacteria (GNB) and the increased use of carbapenems to treat infections unresponsive to the penicillins and cephems (cephalosporins and cephamycins), carbapenemase-producing and carbapenem-resistant GNB are becoming common worlwide 1,2. Thus, carbapenems, which are last-resort antibiotics, are being replaced by colistin, a polymyxin that was discovered many years ago but shelved over nephrotoxicity and neurotoxicity concerns 2,3. Resistance to colistin, due to its increased use to treat carbapenem-resistant GNB, has also emerged 2-4. Beginning from 2016, colistin resistance, which hitherto was mediated my chromosomal mutations, was found to be also mediated by plasmid-borne mcr genes 5,6. The high fatalities caused by colistin and/or carbapenem-resistant GNB infections have led to their being categorised as high-priority pathogens by the WHO 7,8. The clinical importance of colistin-resistant GNB (CR-GNB) necessitates an efficient diagnostic protocol for quickly and efficiently detecting them to preempt further escalation and mortalities 9 .

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“…Early detection of MCRpositive bacteria would help setting policy in preventing further spread and providing appropriate antimicrobial therapy. Despite of traditional loop-mediated isothermal amplification, multiplex-LAMP assay has been developed and successfully detection ofmcr-1 to mcr-5in colistin-resistant bacteria [29,30]. MLST was used to detect and characterise ESBL-producing E. coli that express mcr-1 obtained from dairy cows in China [18].…”

Section: Discussionmentioning

confidence: 99%

Multiplex PCR for detection of MCR genes in clinical fecal samples

Xiang

Hu²,

et al. 2021

E3S Web Conf.

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Plasmid-mediated colistin-resistance genes have been reported worldwide in recent years. A multiplex polymerase chain reaction (Multi-PCR) protocol was developed to detect transferable colistinresistance genes (mcr-1 to mcr-6) in Enterobacteria for clinical laboratory purposes.The authors first designed six new primer pairs to amplify mcr-1 to mcr-6 gene products to achieve stepwise separation of amplicons between 87 to 216 bp,then divided these primers into two subgroups with the assistance of a pair of universal primers for the detection of currently described mcr genes and their variants in Enterobacteria. The protocol was validated by testing 29 clinical isolates of Escherichia coli of human origin, each well characterised and prospectively validated. The Multi-PCR assay showed full concordance with whole-genome sequence data and displayed higher sensitivity and 100% specificity. The assay could detect all variants of the various mcr alleles described. It was able to detect mcr-3 and mcr-4 as singletons or in combination. This type of test is critical for the epidemiological surveillance of plasmid-encoded resistance in limited resources conditions, and this method allows rapid identification of mcr-positive bacteria and overcomes the challenges of phenotypic detection of colistin resistance.

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Assessment of two selective agar media to isolate colistin-resistant bovine Escherichia coli: Correlation with minimal inhibitory concentration and presence of mcr genes

Cited by 15 publications

References 22 publications

Comparative Evaluation of CHROMagar COL-APSE, MicroScan Walkaway, ComASP Colistin, and Colistin MAC Test in Detecting Colistin-resistant Gram-Negative Bacteria

Comparative Evaluation of CHROMagar COL-APSE, MicroScan Walkaway, ComASP Colistin, and Colistin MAC Test in Detecting Colistin-resistant Gram-Negative Bacteria

Comparative Evaluation of CHROMagar COL-APSE, MicroScan Walkaway, ComASP Colistin, and Colistin MAC Test in Detecting Colistin-resistant Gram-Negative Bacteria

Multiplex PCR for detection of MCR genes in clinical fecal samples

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