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Carbapenemase Producing Klebsiella pneumoniae (KPC): What Is the Best MALDI-TOF MS Detection Method
Lukáš Hleba1, Miroslava Hlebová2, Anton Kováčik3
1Institute of Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Trieda Andreja Hlinku 2, 949 76 Nitra, Slovakia.
Abstract:
Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria is a group of highly dangerous antibiotic resistant Gram-negative Enterobacteriaceae. They cause infections associated with significant morbidity and mortality. Therefore, the rapid detection of KPC-producing bacteria plays a key role in clinical microbiology. Matrix assisted laser desorption/ionization time-of- flight (MALDI-TOF) is a rapidly evolving technology that finds application in various clinical, scientific, and industrial disciplines. In the present study, we demonstrated three different procedures of carbapenemase-producing K. pneumoniae (KPC) detection. The most basic model of MALDI-TOF instrument MS Microflex LT was used, operating in the linear ion-positive mode, commonly used in modern clinical laboratories. The first procedure was based on indirect monitoring of carbapenemase production with direct detection of hydrolyzed carbapenem antibiotic degradation products in the mass spectrum. The second procedure was based on direct detection of blaKPC accompanying peak with an 11,109 Da in the mass spectrum of carbapenemase-producing K. pneumoniae (KPC), which represents the cleaved protein (pKpQIL_p019) expressed by pKpQIL plasmid. In addition, several unique peaks were detected in the carbapenemase-producing K. pneumoniae (KPC) mass spectrum. The third procedure was the identification of carbapenemase-producing K. pneumoniae (KPC) based on the protein fingerprint using local database created from the whole mass spectra. By comparing detection procedures, we determined that the third procedure was very fast and relatively easy. However, it requires previous verification of carbapenemase-producing K. pneumoniae (KPC) using other methods as genetic blaKPC identification, detection of carbapenem degradation products, and accompanying peak with 11,109 Da, which represents cleaved pKpQIL_p019 protein expressed by pKpQIL plasmid. Detection of carbapenemase-producing K. pneumoniae using MALDI-TOF provides fast and accurate results that may help to reduce morbidity and mortality in hospital setting when applied in diagnostic situations.
Insights
Rapidly detect Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria using MALDI-TOF. This method offers fast, accurate results to reduce patient morbidity and mortality in clinical settings.
Area of Science:
- Clinical Microbiology
- Mass Spectrometry
- Antimicrobial Resistance
Background:
- Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria are highly dangerous, causing severe infections with significant morbidity and mortality.
- Rapid detection of KPC-producing bacteria is crucial for effective clinical management and infection control.
- Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) is a rapidly advancing technology with diverse applications.
Purpose of the Study:
- To demonstrate and compare three distinct procedures for the rapid detection of KPC-producing Klebsiella pneumoniae using MALDI-TOF.
- To evaluate the efficiency and accuracy of MALDI-TOF based methods for identifying carbapenemase-producing Enterobacteriaceae in a clinical microbiology context.
Main Methods:
- Utilized a basic MALDI-TOF instrument (MS Microflex LT) operating in linear ion-positive mode.
- Developed three detection procedures: indirect monitoring of carbapenem hydrolysis products, direct detection of the blaKPC gene product (11,109 Da peak), and protein fingerprinting using a local database.
- Compared the speed, ease of use, and accuracy of the three developed MALDI-TOF detection methods.
Main Results:
- Identified unique mass spectral peaks associated with KPC-producing Klebsiella pneumoniae, including a peak at 11,109 Da representing cleaved pKpQIL_p019 protein.
- Protein fingerprinting using a local database proved to be very fast and relatively easy for KPC identification.
- Verification of the protein fingerprinting method requires confirmation using genetic blaKPC identification or detection of carbapenem degradation products.
Conclusions:
- MALDI-TOF mass spectrometry offers a fast and accurate method for detecting carbapenemase-producing Klebsiella pneumoniae.
- The protein fingerprinting approach, while rapid, necessitates validation by other methods for definitive identification.
- Implementing MALDI-TOF detection in diagnostic settings can significantly aid in reducing hospital-acquired infections and improving patient outcomes.
