Annals of clinical microbiology and antimicrobials · 2025
Exploring New Delhi Metallo Beta Lactamases in Klebsiella pneumoniae and Escherichia coli: genotypic vs. phenotypic insights.
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Ain Noor Ul, Elton Linzy, Sadouki Zahra, McHugh Timothy D, Riaz Saba
Аннотация
BACKGROUND: Carbapenemase-producing Enterobacterales pose a serious clinical threat, particularly in high-burden settings of carbapenem-resistant Escherichia coli and Klebsiella pneumoniae (CREK), where rapid detection tools are essential to aid patient management. In this study, we focused on blaNDM, the most frequently reported carbapenemase in the region, and evaluated a combined phenotypic (lateral flow) and genotypic (PCR and WGS) approach for its detection. This research underscores the utility of lateral flow assays as a practical alternative to resource-intensive genotypic methods, offering a scalable solution for settings with limited laboratory capacity.
METHOD: One hundred seventy-seven extensively drug-resistant strains were characterized using MALDI-TOF. Isolates were analyzed to detect Carbapenem-resistant Escherichia coli and Klebsiella pneumoniae (CREK) using disk diffusion, MIC test, and PCR targeting blaNDM. Antibiotic susceptibility patterns were analyzed and visualized using single-linkage hierarchical clustering, with results displayed on a permuted heat map. Immunochromatographic assay, RESIST-5 O.K.N.V.I (Coris Bioconcept®) was used for CREK isolates [(n = 17), positive and negative)] and Oxford Nanopore Sequencing was conducted on subsets [(n = 5) blaNDM-positive co-producers of blaNDM and blaOXA, and (n = 2) blaNDM-negative blaOXA producers) to evaluate the reliability of phenotypic and genotypic tests.
RESULT: Most of the XDR strains (90%) were CREK, with K. pneumoniae (71.2%) more prevalent than E. coli (28.7%) (p < 0.05). All CREK strains exhibited complete resistance (100%) to multiple antibiotics with 66% showing sensitivity to levofloxacin. Furthermore, K. pneumoniae (57.8%) had higher blaNDM gene prevalence than E. coli (36.9%). Among blaNDM-positive CREK, lateral flow assay revealed approximately half of each bacteria type co-produced blaOXA (E.coli, 52.9%), and (K. pneumoniae, 47%). For blaNDM-negative strains, blaOXA was more prevalent in K. pneumoniae (82.35%) than E. coli (41%) (p < 0.05). Comparing phenotypic to genotypic assays, E. coli showed 100% (CI 80.49 - 100%) sensitivity and specificity with a high Kappa agreement coefficient (0.91) (CI 95% 0.661-1, p < 0.01), whereas K. pneumoniae assays had lower sensitivity and specificity (40%) (CI 5.27 - 85.34%), with a lower Kappa agreement coefficient (0.20) (CI 95% 0.104-0.298, p < 0.01).
CONCLUSION: This study demonstrates the value of the RESIST-5 O.K.N.V.I. lateral flow assay as a rapid and reliable diagnostic tool for detecting blaNDM in Escherichia coli, with strong agreement to PCR and WGS. While performance for Klebsiella pneumoniae was lower, the assay offers a practical alternative in resource-limited settings, aiding antimicrobial stewardship and improving diagnostic capacities in high-burden regions.
METHOD: One hundred seventy-seven extensively drug-resistant strains were characterized using MALDI-TOF. Isolates were analyzed to detect Carbapenem-resistant Escherichia coli and Klebsiella pneumoniae (CREK) using disk diffusion, MIC test, and PCR targeting blaNDM. Antibiotic susceptibility patterns were analyzed and visualized using single-linkage hierarchical clustering, with results displayed on a permuted heat map. Immunochromatographic assay, RESIST-5 O.K.N.V.I (Coris Bioconcept®) was used for CREK isolates [(n = 17), positive and negative)] and Oxford Nanopore Sequencing was conducted on subsets [(n = 5) blaNDM-positive co-producers of blaNDM and blaOXA, and (n = 2) blaNDM-negative blaOXA producers) to evaluate the reliability of phenotypic and genotypic tests.
RESULT: Most of the XDR strains (90%) were CREK, with K. pneumoniae (71.2%) more prevalent than E. coli (28.7%) (p < 0.05). All CREK strains exhibited complete resistance (100%) to multiple antibiotics with 66% showing sensitivity to levofloxacin. Furthermore, K. pneumoniae (57.8%) had higher blaNDM gene prevalence than E. coli (36.9%). Among blaNDM-positive CREK, lateral flow assay revealed approximately half of each bacteria type co-produced blaOXA (E.coli, 52.9%), and (K. pneumoniae, 47%). For blaNDM-negative strains, blaOXA was more prevalent in K. pneumoniae (82.35%) than E. coli (41%) (p < 0.05). Comparing phenotypic to genotypic assays, E. coli showed 100% (CI 80.49 - 100%) sensitivity and specificity with a high Kappa agreement coefficient (0.91) (CI 95% 0.661-1, p < 0.01), whereas K. pneumoniae assays had lower sensitivity and specificity (40%) (CI 5.27 - 85.34%), with a lower Kappa agreement coefficient (0.20) (CI 95% 0.104-0.298, p < 0.01).
CONCLUSION: This study demonstrates the value of the RESIST-5 O.K.N.V.I. lateral flow assay as a rapid and reliable diagnostic tool for detecting blaNDM in Escherichia coli, with strong agreement to PCR and WGS. While performance for Klebsiella pneumoniae was lower, the assay offers a practical alternative in resource-limited settings, aiding antimicrobial stewardship and improving diagnostic capacities in high-burden regions.