Investigation of the presence of carbapenemases in carbapenem-resistant Klebsiella pneumoniae strains by MALDI-TOF MS (matrix-assisted laser desorption ionization-time of flight mass spectrometry) and comparison with real-time PCR method |
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| Institution: | 1. Kastamonu Research and Training Hospital, Department of Medical Microbiology, Kastamonu, Turkey;2. Ege University Faculty of Medicine, Department of Medical Microbiology, İzmir, Turkey;1. Indian Council of Agricultural Research -National Institute of Veterinary Epidemiology and Disease Informatics (ICAR-NIVEDI), Yelahanka, Bengaluru, 560 064, Karnataka, India;2. Indian Council of Medical Research-National Institute of Occupational Health (ICMR-NIOH), Meghaninagar, Ahmedabad, 380 016, Gujarat, India;3. Department of Microbiology, JSS Medical College, Mysuru, 570 015, Karnataka, India;4. Department of Veterinary Public Health & Epidemiology, Nagpur Veterinary College, Seminary Hills, Nagpur, 440 006 Maharashtra, India;1. Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, 110070, India;2. Department of Research, Institute of Liver and Biliary Sciences, New Delhi, 110070, India;3. Department of Hepatology, Institute of Liver and Biliary Sciences, New Delhi, 110070, India;1. Department of Pharmaceutical Biotechnology, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran;2. Students Research Committee, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran;3. Department of Infectious Diseases, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran;4. Department of Clinical Pharmacy and Pharmacy Practice, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran;5. Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;1. Department of Clinical Microbiology & Immunology, Sir Ganga Ram Hospital, Rajinder Nagar, New Delhi, 110060, India;2. Critical Care & Emergency Medicine, Sir Ganga Ram Hospital, Rajinder Nagar, New Delhi, 110060, India;3. Department of Internal Medicine, Sir Ganga Ram Hospital, Rajinder Nagar, New Delhi, 110060, India;1. Department of Infectious Diseases, Sterling Hospital, Ahmedabad, 380052, India;2. Division of Infectious Diseases, Department of Internal Medicine, Morsani College of Medicine, University of South Florida, Tampa, 33612, USA;3. Infectious Diseases Consultant, 405, AXIS Business Space, Nanpura, Surat, 395001, India;4. Clinical Microbiologist, Abha Laboratory Pvt Ltd, 2nd Floor, Rajratna Chambers, Dabgharwad, Bhagal, Surat, India;1. Department of Microbiology, ESIC Medical College & Hospital, Sanathnagar, Hyderabad, India;2. Department of Microbiology, All India Institute of Medical Sciences, Gorakhpur, India;3. Department of Biochemistry, ESIC Medical College & Hospital, Sanathnagar, Hyderabad, India |
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| Abstract: | PurposeWe aimed to develop a new procedure for rapid detection of the carbapenemase activity using MALDI-TOF MS, and to determine the sensitivity and specificity of the method. Also, we aimed to determine the distribution of carbapenemase genes among the K.pneumoniae strains isolated in our hospital using real-time PCR.MethodBetween January 2017–February 2019; K. pneumoniae strains(n = 74) isolated from blood culture samples were included. Klebsiella pneumoniae NCTC 13438 was used as a positive control and Escherichia coli ATCC 25922 as a negative control. First, Imipenem, meropenem, and ertapenem MIC values of strains were determined. Then blaKPC, blaOXA-48, and/or blaNDM genes were investigated with PCR. Carbapenemase activity was investigated in strains with the newly developed method using MALDI-TOF MS. The performance of the new method was evaluated for both the second and fourth hours of the incubation period.ResultsWhile 65 strains were found resistant to tested carbapenems, nine of them were susceptible. Of the 65 resistant strains, 57 had blaOXA-48, 15 had blaNDM, and four had blaKPC genes. BlaOXA-48 and blaNDM genes were detected together in 11 strains. BlaOXA-48, blaNDM, and blaKPC genes were not detected in any of the susceptible strains. The sensitivity and specificity of MALDI-TOF MS at the second hour were 83.1% and 100%, respectively. At the fourth hour, the sensitivity and specificity of MALDI-TOF MS were 100%. No false-positive results were observed.ConclusionThe sensitivity of the method at the fourth hour was better than the second hour. The false-negative results observed in the second hour disappeared when the incubation period was extended to 4 h. MALDI-TOF MS which is still under development is a fast, cost-effective, promising method for the detection of carbapenemase activity. |
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| Keywords: | MALDI TOF MS Carbapenemase activity Ertapenem Ertapenem hydrolysis assay |
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