Identification and elimination of the clinically relevant multi-resistant environmental bacteria Ralstonia insidiosa in primary cell culture
In times of spreading multidrug-resistant bacteria, species identification and decontamination of cell cultures can be challenging. Here, we describe a mobile cell culture contaminant with “black dot”-like microscopic appearance in newly established irreplaceable hybridoma cell l...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
17 October 2020
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| In: |
Microorganisms
Year: 2020, Volume: 8, Issue: 10 |
| ISSN: | 2076-2607 |
| DOI: | 10.3390/microorganisms8101599 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.3390/microorganisms8101599 Verlag, lizenzpflichtig, Volltext: https://www.mdpi.com/2076-2607/8/10/1599 
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| Author Notes: | Dennis Nurjadi, Sébastien Boutin, Katja Schmidt, Melinda Ahmels and Daniel Hasche |
| Summary: | In times of spreading multidrug-resistant bacteria, species identification and decontamination of cell cultures can be challenging. Here, we describe a mobile cell culture contaminant with “black dot”-like microscopic appearance in newly established irreplaceable hybridoma cell lines and its identification. Using 16S rRNA gene sequencing, species-specific PCRs, whole genome sequencing (WGS), and MALDI-TOF mass spectrometry, the contaminant was identified as the ubiquitous environmental and clinically relevant Gram-negative bacterium Ralstonia insidiosa (R. insidiosa), a strong biofilm producer. Further characterizations by transmission electron microscopy (TEM) and biochemical API test were not conclusive. Whole genome sequencing of our R. insidiosa isolate revealed numerous drug-resistance determinants. Genome-wide comparison to other Ralstonia species could not unambiguously designate our isolate to R. insidiosa (<95% average nucleotide identity) suggesting a potential novel species or subspecies, closely related to R. insidiosa and R. pickettii. After determining the antibiotic susceptibility profile, the hybridoma cell culture was successfully decontaminated with ciprofloxacin without affecting antibody production. |
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| Item Description: | Gesehen am 23.10.2020 |
| Physical Description: | Online Resource |
| ISSN: | 2076-2607 |
| DOI: | 10.3390/microorganisms8101599 |