It is crucial to identify Enterobacterales that produce carbapenemase to treat and manage hospital infections. The suggested techniques for their identification need a lengthy wait, technical knowledge, and training. Lateral flow immunoassays (LFIAs) provide a solution to these requirements. Thus, this study compared LFIA with phenotypic and genotypic tests for carbapenemase-producing bacteria. Fifty clinical isolates of carbapenem-resistant superbugs were examined. KPC, VIM, NDM, IMP, and OXA-48-like enzymes were evaluated and compared with phenotypic tests and LFIA. Regarding the phenotypic characteristics, the mCIM was positive in 37/50 (74%), and the eCIM was positive in 21/50 (42%). Regarding using LFIA, 41 out of the total isolates (82%) gave a positive red line with one or more of the tested genes. The most frequently detected gene was bla_NDM (27/50 (54%)), and the least detected one was bla_IMP (14/50 (28%)), which was in accordance with the PCR results. While investigating the accuracy of LFIA vs. PCR, it was found that LFIA had 100% sensitivity in the detection of the bla_NDM and bla_OXA genes, with 85.2% and 91.4% specificity, respectively, while for the bla_IMP, bla_KPC, and bla_VIM genes, the values were 91.7% and 92.1%, 94.1% and 90.9%, and 95.5% and 89.3%, respectively. The overall accuracy of LFIA ranged from 92 to 94%. Our comparison with molecular assays revealed remarkable agreement, so we propose that this test might be utilized as a supplementary tool.

To improve clinical decision-making about Carbapenem-resistant Gram-negative bacteria (CR-GNB) infections and halt the spread of resistant microbes, quicker and less expensive diagnostic techniques are required. Thus, the purpose of this study was to thoroughly evaluate the diagnostic efficiency (sensitivity, specificity, and concordance) of direct-from-specimen multiplex lateral flow immunoassay (LFIA) across diverse raw clinical specimens and pathogen types from critically sick patients. A total of 300 non-duplicate samples were tested to detect CR-GNB. Five major Carbapenemase genes were detected directly from the specimen using carbapenem-resistant K.N.I.V.O. detection K-Set and from culture using culture-enhanced multiplex PCR. Turnaround time (TAT) of each method was calculated. The direct LFIA revealed 100% specificity for NDM, KPC, and IMP enzymes in all tested clinical matrices (blood, urine, and respiratory samples). The study demonstrated 100% sensitivity and specificity with perfect categorical agreement (κ = 1.000) for the blaKPC in the Klebsiella pneumoniae and for blaOXA-48 and blaIMP in the Acinetobacter baumannii; however, sensitivity of blaVIM was significantly diminished across all isolates and samples. TAT decreased significantly (p < 0.001) from 30 to 70 h to about 50 min. The tested direct LFIA facilitates the prompt enhancement of lifesaving tailored antibiotic treatment for severe illnesses.