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. 2022 Feb 11:12:761328.
doi: 10.3389/fcimb.2022.761328. eCollection 2022.

Methodology Establishment and Application of VITEK Mass Spectrometry to Detect Carbapenemase-Producing Klebsiella pneumoniae

Haoyun Lin  1 , Zhen Hu  1 , Jinsong Wu  1 , Yuemei Lu  1 , Jine Chen  1 , Wenyuan Wu  1
Affiliations

Methodology Establishment and Application of VITEK Mass Spectrometry to Detect Carbapenemase-Producing Klebsiella pneumoniae

Haoyun Lin et al. Front Cell Infect Microbiol. .

Abstract

The ability of VITEK mass spectrometry (MS) in detection of bacterial resistance is currently under exploration and evaluation. In this study, we developed and validated a VITEK MS method to rapidly test carbapenemase-producing Klebsiella pneumoniae (CPKP). Solvents, antibiotic concentrations, crystal conditions and times, centrifugation speeds, and other factors were optimized to design a rapid sample pretreatment process for CPKP detection by VITEK MS. The related parameters of the mass spectrum were adjusted on the instrument to establish an CPKP detection mode. 133 clinically isolated strains of CPKP in the microbiology laboratory at the Shenzhen People's Hospital from 2004 to 2017 were selected for accuracy evaluation. The fresh suspected strains from the microbiology laboratory in 2020 were used to complete the clinical verification. Two antibiotics, meropenem (MEM) and imipenem (IPM), were used as substrates. These two substrates were incubated with suspected CPKP, and the results were obtained by VITEK MS detection. Using this method, different types of CPKP showed different detection results and all the CPKP strains producing KPC-2 and IMP-4 carbapenemase were detected by VITEK MS. Thus, VITEK MS can be used for rapid detection of CPKP, especially for some common types of CPKP. This method provides high accuracy and speed of detection. Combined with its cost advantages, it can be intensely valuable in clinical microbiology laboratories after the standard operating procedures are determined.

Keywords: IPM; MALDI-TOF MS; MEM; VITEK MS; carbapenemase-producing Klebsiella pneumoniae; hydrolysis experiment; imipenem; meropenem.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Experimental conditions for specimen pretreatment of detecting CPKP based on VITEK MS. (A) List of specific experimental parameters. (B) The parameters of the MEM mass spectrum under different solvent conditions. (C) The parameters of the MEM mass spectrum under different MEM concentrations. (D) The parameters of the MEM mass spectrum under different dry conditions of co-crystalline film formation. (E) The parameters of the MEM mass spectrum under different microbial concentration conditions.
Figure 2
Figure 2
Changes of peak mass spectra of MEM or IPM at different incubation times. (A) The peak value of the MEM mass spectrum at different incubation times. (B) The peak value of IPM mass spectrum at different incubation times.
Figure 3
Figure 3
Flowchart of CPKP detection by VITEK MS.
Figure 4
Figure 4
Detection results of CPKP were obtained by VITEK MS. (A) The changes of the mass spectrum of carbapenemase were detected when MEM was used as a substrate for hydrolysis. NDM-1* represents that NDM-1 genotype CPKP was not detected by VITEK MS. (B) The changes of the mass spectrum of carbapenemase were detected when IPM was used as a substrate for hydrolysis.
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