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Detection of K. pneumoniae Hospital-Acquired Strains That Produce Carbapenemases in Thrace Tertiary Hospital
Anastasia Vezyridou1, Aikaterini Skeva1, Ioanna Alexandropoulou1
1Laboratory of Microbiology, Department of Medicine, Democritus University of Thrace, Dragana, 69100 Alexandroupolis, Greece.
Abstract:
In recent decades, the problem of resistant strains, which present resistance to different types of antimicrobials, has increased. Klebsiella pneumoniae is one of the most important species that exhibits an acquired resistance phenotype to at least one agent in three or more classes of antimicrobials and is thus characterized as a multidrug-resistant bacterium (MDR). 98 nosocomial strains of K. pneumoniae were isolated during the pre-COVID-19 period, and more specifically, from February 2015 to March 2019, were analyzed for the detection of class A, D, and B carbapenemase genes. The existence of KPC, OXA-48 like, IMP, VIM, and NDM carbapenemases has been examined. The immunochromatography showed that NDM carbapenemases are more frequently detected in the samples, reaching a percentage of 30.7%, while correspondingly the percentage for VIM carbapenemases was 7.68% among the strains with resistant phenotypes. No strain with carbapenemase IMP was found. Real-time multiplex polymerase chain reaction (PCR) showed, in contrast to immunochromatography kits, that a high percentage of bacterial isolates (94.26%) carry NDM and VIM carbapenemase genes, while no IMP carbapenemase genes were detected. Regarding the KPC enzymes, the immunochromatography kits showed that KPC positive strains are reaching 53.1%, and OXA-48 positive strains are reaching 3.1% among the strains with resistant phenotypes. Real-time multiplex polymerase chain reaction revealed a much higher percentage of 89.6% KPC positive isolates and a percentage of 14.6% OXA-48 carbapenemase producers. The aforementioned results indicate the dominance of the Multiplex Real-Time PCR as a "gold standard" method. This study could not fully support the usefulness of rapid immunochromatographic tests as a fast and useful diagnostic tool in the laboratory daily routine, as per the results of previous studies. Thus, more studies need to be conducted in this field to introduce these rapid tests safely into the daily laboratory workflow as a screening tool. Additionally, this study underlines the predominance of KPC enzymes from clinical isolates of ICUs and a significant shift over the OXA-48 like enzymes that are not limited to the ICU environment.
Insights
Multidrug-resistant Klebsiella pneumoniae strains are a growing concern. Multiplex Real-Time PCR is superior to immunochromatography for detecting carbapenemase genes like KPC, NDM, and VIM in clinical isolates.
Area of Science:
- Clinical microbiology
- Molecular diagnostics
- Antimicrobial resistance
Background:
- Rising prevalence of multidrug-resistant (MDR) bacteria, particularly Klebsiella pneumoniae.
- Carbapenemase-producing Enterobacteriaceae (CPE) pose a significant global health threat.
- Nosocomial infections caused by MDR K. pneumoniae necessitate accurate and rapid detection methods.
Purpose of the Study:
- To compare the diagnostic performance of immunochromatography and Real-Time Multiplex PCR for detecting carbapenemase genes in K. pneumoniae.
- To investigate the prevalence of specific carbapenemase genes (KPC, OXA-48 like, IMP, VIM, NDM) in nosocomial K. pneumoniae isolates.
- To evaluate the utility of rapid immunochromatographic tests in routine laboratory diagnostics.
Main Methods:
- Analysis of 98 nosocomial K. pneumoniae strains isolated between February 2015 and March 2019.
- Detection of carbapenemase genes (KPC, OXA-48 like, IMP, VIM, NDM) using immunochromatography and Real-Time Multiplex PCR.
- Comparison of sensitivity and specificity between the two detection methods.
Main Results:
- Real-Time Multiplex PCR identified a significantly higher prevalence of NDM (94.26%) and VIM (94.26%) genes compared to immunochromatography.
- Immunochromatography detected KPC (53.1%) and OXA-48 like (3.1%) genes, while PCR revealed higher rates (KPC: 89.6%, OXA-48 like: 14.6%).
- No IMP carbapenemase genes were detected by either method. NDM was most frequent by immunochromatography (30.7%).
Conclusions:
- Multiplex Real-Time PCR is the gold standard for detecting carbapenemase genes, demonstrating superior sensitivity.
- Rapid immunochromatographic tests showed limitations and require further validation before routine use.
- KPC enzymes are predominant in clinical isolates from ICUs, with OXA-48 like enzymes showing a broader distribution.
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