The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytoca
IntroductionKlebsiella pneumoniae and K. oxytoca are members of Enterobacteriaceae. They are Gram-negative, non-motile rods that are ubiquitous in the environment and part of the human intestinal microbiota. These opportunistic pathogens may cause pneumonia and urinary tract infections. Klebsiella s...
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Frontiers Media S.A.
2025-02-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1514643/full |
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| author | Agata Palusiak Anna Maciejewska Jolanta Łukasiewicz |
| author_facet | Agata Palusiak Anna Maciejewska Jolanta Łukasiewicz |
| author_sort | Agata Palusiak |
| collection | DOAJ |
| description | IntroductionKlebsiella pneumoniae and K. oxytoca are members of Enterobacteriaceae. They are Gram-negative, non-motile rods that are ubiquitous in the environment and part of the human intestinal microbiota. These opportunistic pathogens may cause pneumonia and urinary tract infections. Klebsiella species are genetically and biochemically similar; therefore, it is important to find reliable methods for their differentiation.MethodsThis study presents the results of biochemical assays, PCR, and MALDI-TOF mass spectrometry (MS) performed on 35 Klebsiella isolates obtained from the urine of patients from central Poland.ResultsAmong biochemical methods, the indole test demonstrated the highest discriminatory power, whereas the determination of growth at 10°C was the least effective. For all strains biochemically identified as K. pneumoniae, a 108-bp amplicon was detected, indicating the presence of the rpoB gene in their genome. Only 12 K. oxytoca isolates produced a product of the pehX gene. All tested strains were analyzed using the MALDI-TOF Biotyper, which confirmed, with high-quality scores, their identification based on api 20E and indole tests. Strain 0.011 was identified as Raoultella ornithinolytica.ConclusionMALDI-TOF MS analysis proved to be the most reliable method for identifying K. oxytoca and K. pneumoniae, with the potential for phylogroup differentiation. |
| format | Article |
| id | doaj-art-a494c58bf8e2468bb0d941255bef60b7 |
| institution | DOAJ |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj-art-a494c58bf8e2468bb0d941255bef60b72025-08-20T02:45:30ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-02-011610.3389/fmicb.2025.15146431514643The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytocaAgata Palusiak0Anna Maciejewska1Jolanta Łukasiewicz2Department of Biology of Bacteria, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, PolandLaboratory of Microbial Immunochemistry and Vaccines, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, PolandLaboratory of Microbial Immunochemistry and Vaccines, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, PolandIntroductionKlebsiella pneumoniae and K. oxytoca are members of Enterobacteriaceae. They are Gram-negative, non-motile rods that are ubiquitous in the environment and part of the human intestinal microbiota. These opportunistic pathogens may cause pneumonia and urinary tract infections. Klebsiella species are genetically and biochemically similar; therefore, it is important to find reliable methods for their differentiation.MethodsThis study presents the results of biochemical assays, PCR, and MALDI-TOF mass spectrometry (MS) performed on 35 Klebsiella isolates obtained from the urine of patients from central Poland.ResultsAmong biochemical methods, the indole test demonstrated the highest discriminatory power, whereas the determination of growth at 10°C was the least effective. For all strains biochemically identified as K. pneumoniae, a 108-bp amplicon was detected, indicating the presence of the rpoB gene in their genome. Only 12 K. oxytoca isolates produced a product of the pehX gene. All tested strains were analyzed using the MALDI-TOF Biotyper, which confirmed, with high-quality scores, their identification based on api 20E and indole tests. Strain 0.011 was identified as Raoultella ornithinolytica.ConclusionMALDI-TOF MS analysis proved to be the most reliable method for identifying K. oxytoca and K. pneumoniae, with the potential for phylogroup differentiation.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1514643/fullbiochemical methodsKlebsiella oxytocaKlebsiella pneumoniaeMALDI-TOF mass spectrometryPCRspecies identification |
| spellingShingle | Agata Palusiak Anna Maciejewska Jolanta Łukasiewicz The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytoca Frontiers in Microbiology biochemical methods Klebsiella oxytoca Klebsiella pneumoniae MALDI-TOF mass spectrometry PCR species identification |
| title | The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytoca |
| title_full | The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytoca |
| title_fullStr | The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytoca |
| title_full_unstemmed | The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytoca |
| title_short | The results of polymerase chain reaction and MALDI-TOF mass spectrometry versus phenotypic distinction between Klebsiella pneumoniae and Klebsiella oxytoca |
| title_sort | results of polymerase chain reaction and maldi tof mass spectrometry versus phenotypic distinction between klebsiella pneumoniae and klebsiella oxytoca |
| topic | biochemical methods Klebsiella oxytoca Klebsiella pneumoniae MALDI-TOF mass spectrometry PCR species identification |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1514643/full |
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