Globally, Klebsiella pneumoniae (K. pneumoniae) has been identified as a serious source of infections. The objectives of our study were to investigate the prevalence of multidrug-resistant (MDR) K. pneumoniae in Tanta University Hospitals, Gharbia Governorate, Egypt; characterize their carbapenem resistance profiles; and identify their different capsular serotypes. We identified and isolated 160 (32%) K. pneumoniae from 500 different clinical samples, performed antimicrobial susceptibility testing, and then used multiplex PCR to detect carbapenemase genes and capsular serotypes K1, K2, K3, K5, K20, K54, and K57. We detected phenotypic carbapenem resistance in 31.3% (50/160) of the isolates; however, molecular assays revealed that 38.75% (62/160) of isolates were carrying carbapenemase-encoding genes. Generally, bla_OXA-48 was the prevalent gene (15.5%), followed by bla_VIM (15%), bla_IMP (7.5%), bla_KPC (4%), and bla_NDM (3.8%). Bla_VIM and bla_OXA-48 correlated with phenotypic resistance in 91.67% and 88% of the isolates that harbored them, respectively. Capsular typing showed that the most prevalent pathotype was K1 (30.6%), followed by K57 (24.2%), K54 (19.35%), K20 (9.67%), and K2 (6.45%). A critical risk to community health is posed by the high incidence of multidrug-resistant (MDR) virulent K. pneumoniae isolates from our hospital, and our study examines this pathogen’s public health and epidemiological risks.

Methicillin-resistant in Staphylococci is a serious public health issue. It is mostly encoded by the mecA gene. The mecC gene is a new mecA analog responsible for resistance to methicillin in some Staphylococcal clinical isolates. This mecC gene is still underestimated in Egypt. The aim of the current study was to detect mecA and mecC genes in clinical Staphylococci isolates from a tertiary care university hospital in Egypt compared to the different phenotypic methods. A total of 118 Staphylococcus aureus (S. aureus) and 43 coagulase-negative Staphylococci (CoNS) were identified from various hospital-acquired infections. Methicillin resistance was identified genotypically using the PCR technique and phenotypically using the cefoxitin disc diffusion test, oxacillin broth microdilution and the VITEK2 system in all Staphylococcal isolates. The mecA gene was detected in 82.2% of S. aureus and 95.3% of CoNS isolates, while all of the isolates tested negative for the mecC gene. Interestingly, 30.2% of CoNS isolates showed the unique character of inducible oxacillin resistance, being mecA-positive but oxacillin-susceptible (OS-CoNS). The dual use of genotypic and phenotypic methods is highly recommended to avoid missing any genetically divergent strains.

Hypervirulent Klebsiella pneumoniae is a highly pathogenic variant of Klebsiella pneumonae, which represents a global public health issue because it is very virulent and spreads easily. The objectives of this study were to assess the predominance of hvKp among health care-associated infections in intensive care units of Tanta University Hospital and to compare hvKp with classical K. pneumoniae (cKp) in terms of antibiotic resistance, virulence, and molecular features. The study included 300 patients suffering from HAIs from different ICUs of Tanta University Hospitals. K. pneumoniae isolates were identified and subjected to string testing and antibiotic susceptibility testing, and the tissue culture assay for biofilm formation and polymerase chain reaction (PCR) tests were performed for the identification of capsular genes (K1, K2, K57) and virulence genes (rmpA, rmpA2, iuc A). Fifty-seven K. pneumonaie isolates were isolated. A total of 21 (36.8%) of them were hvKp and 36 (63.15%) were cKp. Significantly higher antibiotic resistance was detected in the cKp group. There was a significant difference between biofilm formation between cKp and hvKp isolates (p < 0.004*). iucA, rmpA2, and K1 genes were significantly associated with hvKp. The string test shows 100% sensitivity and negative predictive value for the detection of hvKp. Consequently, using the string test alone for the screening of hvKp is required. However, combining aerobactin-positive with hypermucoviscous isolates while screening for hvKp is crucial.

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.