Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemases
Abstract The resistance to carbapenems is usually mediated by enzymes hydrolyzing β-lactam ring. Recently, an alternative way of the modification of the antibiotic, a β-lactone formation by OXA-48-like enzymes, in some carbapenems was identified. We focused our study on a deep analysis of OXA-48-lik...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-12-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-26735-5 |
| _version_ | 1828087203002056704 |
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| author | Vendula Studentova Vendula Sudova Ibrahim Bitar Veronika Paskova Jiri Moravec Petr Pompach Michael Volny Petr Novak Jaroslav Hrabak |
| author_facet | Vendula Studentova Vendula Sudova Ibrahim Bitar Veronika Paskova Jiri Moravec Petr Pompach Michael Volny Petr Novak Jaroslav Hrabak |
| author_sort | Vendula Studentova |
| collection | DOAJ |
| description | Abstract The resistance to carbapenems is usually mediated by enzymes hydrolyzing β-lactam ring. Recently, an alternative way of the modification of the antibiotic, a β-lactone formation by OXA-48-like enzymes, in some carbapenems was identified. We focused our study on a deep analysis of OXA-48-like-producing Enterobacterales, especially strains showing poor hydrolytic activity. In this study, well characterized 74 isolates of Enterobacterales resistant to carbapenems were used. Carbapenemase activity was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), liquid chromatography/mass spectrometry (LC–MS), Carba-NP test and modified Carbapenem Inactivation Method (mCIM). As meropenem-derived β-lactone possesses the same molecular weight as native meropenem (MW 383.46 g/mol), β-lactonization cannot be directly detected by MALDI-TOF MS. In the spectra, however, the peaks of m/z = 340.5 and 362.5 representing decarboxylated β-lactone and its sodium adduct were detected in 25 out of 35 OXA-48-like producers. In the rest 10 isolates, decarboxylated hydrolytic product (m/z = 358.5) and its sodium adduct (m/z = 380.5) have been detected. The peak of m/z = 362.5 was detected in 3 strains co-producing OXA-48-like and NDM-1 carbapenemases. The respective signal was identified in no strain producing class A or class B carbapenemase alone showing its specificity for OXA-48-like carbapenemases. Using LC–MS, we were able to identify meropenem-derived β-lactone directly according to the different retention time. All strains with a predominant β-lactone production showed negative results of Carba NP test. In this study, we have demonstrated that the strains producing OXA-48-like carbapenemases showing false-negative results using Carba NP test and MALDI-TOF MS preferentially produced meropenem-derived β-lactone. We also identified β-lactone-specific peak in MALDI-TOF MS spectra and demonstrated the ability of LC–MS to detect meropenem-derived β-lactone. |
| first_indexed | 2024-04-11T05:08:44Z |
| format | Article |
| id | doaj.art-952ed84121a344ee94e6ac05892f69c3 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T05:08:44Z |
| publishDate | 2022-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-952ed84121a344ee94e6ac05892f69c32022-12-25T12:10:26ZengNature PortfolioScientific Reports2045-23222022-12-0112111210.1038/s41598-022-26735-5Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemasesVendula Studentova0Vendula Sudova1Ibrahim Bitar2Veronika Paskova3Jiri Moravec4Petr Pompach5Michael Volny6Petr Novak7Jaroslav Hrabak8Biomedical Center, Faculty of Medicine in Pilsen, Charles UniversityBiomedical Center, Faculty of Medicine in Pilsen, Charles UniversityBiomedical Center, Faculty of Medicine in Pilsen, Charles UniversityBiomedical Center, Faculty of Medicine in Pilsen, Charles UniversityBiomedical Center, Faculty of Medicine in Pilsen, Charles UniversityInstitute of Microbiology of the Czech Academy of Sciences, BIOCEVInstitute of Microbiology of the Czech Academy of Sciences, BIOCEVInstitute of Microbiology of the Czech Academy of Sciences, BIOCEVBiomedical Center, Faculty of Medicine in Pilsen, Charles UniversityAbstract The resistance to carbapenems is usually mediated by enzymes hydrolyzing β-lactam ring. Recently, an alternative way of the modification of the antibiotic, a β-lactone formation by OXA-48-like enzymes, in some carbapenems was identified. We focused our study on a deep analysis of OXA-48-like-producing Enterobacterales, especially strains showing poor hydrolytic activity. In this study, well characterized 74 isolates of Enterobacterales resistant to carbapenems were used. Carbapenemase activity was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), liquid chromatography/mass spectrometry (LC–MS), Carba-NP test and modified Carbapenem Inactivation Method (mCIM). As meropenem-derived β-lactone possesses the same molecular weight as native meropenem (MW 383.46 g/mol), β-lactonization cannot be directly detected by MALDI-TOF MS. In the spectra, however, the peaks of m/z = 340.5 and 362.5 representing decarboxylated β-lactone and its sodium adduct were detected in 25 out of 35 OXA-48-like producers. In the rest 10 isolates, decarboxylated hydrolytic product (m/z = 358.5) and its sodium adduct (m/z = 380.5) have been detected. The peak of m/z = 362.5 was detected in 3 strains co-producing OXA-48-like and NDM-1 carbapenemases. The respective signal was identified in no strain producing class A or class B carbapenemase alone showing its specificity for OXA-48-like carbapenemases. Using LC–MS, we were able to identify meropenem-derived β-lactone directly according to the different retention time. All strains with a predominant β-lactone production showed negative results of Carba NP test. In this study, we have demonstrated that the strains producing OXA-48-like carbapenemases showing false-negative results using Carba NP test and MALDI-TOF MS preferentially produced meropenem-derived β-lactone. We also identified β-lactone-specific peak in MALDI-TOF MS spectra and demonstrated the ability of LC–MS to detect meropenem-derived β-lactone.https://doi.org/10.1038/s41598-022-26735-5 |
| spellingShingle | Vendula Studentova Vendula Sudova Ibrahim Bitar Veronika Paskova Jiri Moravec Petr Pompach Michael Volny Petr Novak Jaroslav Hrabak Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemases Scientific Reports |
| title | Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemases |
| title_full | Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemases |
| title_fullStr | Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemases |
| title_full_unstemmed | Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemases |
| title_short | Preferred β-lactone synthesis can explain high rate of false-negative results in the detection of OXA-48-like carbapenemases |
| title_sort | preferred β lactone synthesis can explain high rate of false negative results in the detection of oxa 48 like carbapenemases |
| url | https://doi.org/10.1038/s41598-022-26735-5 |
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