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      Tsukamurella pulmonis conjunctivitis in patients with an underlying nasolacrimal duct obstruction – report of two cases

      case-report

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          Abstract

          Tsukamurella pulmonis ( Actinobacteria ), a Gram-positive, obligate aerobic and weakly or variably acid-fast bacterium, is an opportunistic pathogen. Here we report two cases of conjunctivitis caused by T. pulmonis . Both patients had a previous history of nasolacrimal duct obstruction (NLDO). Isolation of T. pulmonis was performed on chocolate, tryptic soy blood and Columbia nalidixic agars. After 24 h of incubation, odourless, white-greyish, membrane-like colonies were observed. The VITEK-2 bacterial identifier system failed to identify the species, while Vitek-MS matrix-assisted laser desorption ionization time-of-flight technology, successfully identified the isolate from case 2 but not from case 1. Final identification was verified using 16S rRNA gene sequencing. An antibiogram was performed and according to the results cefazoline in addition to vancomycin eye drops for 5 days, were suggested as a treatment in case 1. In case 2 the infection was ended without treatment. This is the first report of Tsukamurella as a pathogen that causes conjunctivitis in patients with NLDO.

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          Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

          Piseth Seng, Michel Drancourt, Frédérique Gouriet … (2009)
          Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry accurately identifies both selected bacteria and bacteria in select clinical situations. It has not been evaluated for routine use in the clinic. We prospectively analyzed routine MALDI-TOF mass spectrometry identification in parallel with conventional phenotypic identification of bacteria regardless of phylum or source of isolation. Discrepancies were resolved by 16S ribosomal RNA and rpoB gene sequence-based molecular identification. Colonies (4 spots per isolate directly deposited on the MALDI-TOF plate) were analyzed using an Autoflex II Bruker Daltonik mass spectrometer. Peptidic spectra were compared with the Bruker BioTyper database, version 2.0, and the identification score was noted. Delays and costs of identification were measured. Of 1660 bacterial isolates analyzed, 95.4% were correctly identified by MALDI-TOF mass spectrometry; 84.1% were identified at the species level, and 11.3% were identified at the genus level. In most cases, absence of identification (2.8% of isolates) and erroneous identification (1.7% of isolates) were due to improper database entries. Accurate MALDI-TOF mass spectrometry identification was significantly correlated with having 10 reference spectra in the database (P=.01). The mean time required for MALDI-TOF mass spectrometry identification of 1 isolate was 6 minutes for an estimated 22%-32% cost of current methods of identification. MALDI-TOF mass spectrometry is a cost-effective, accurate method for routine identification of bacterial isolates in or =10 reference spectra per bacterial species and a 1.9 identification score (Brucker system). It may replace Gram staining and biochemical identification in the near future.
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            Performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacterial strains routinely isolated in a clinical microbiology laboratory.

            G Prod'hom, A Bizzini, G Greub … (2010)
            Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been introduced in diagnostic microbiology laboratories for the identification of bacterial and yeast strains isolated from clinical samples. In the present study, we prospectively compared MALDI-TOF MS to the conventional phenotypic method for the identification of routine isolates. Colonies were analyzed by MALDI-TOF MS either by direct deposition on the target plate or after a formic acid-acetonitrile extraction step if no valid result was initially obtained. Among 1,371 isolates identified by conventional methods, 1,278 (93.2%) were putatively identified to the species level by MALDI-TOF MS and 73 (5.3%) were identified to the genus level, but no reliable identification was obtained for 20 (1.5%). Among the 1,278 isolates identified to the species level by MALDI-TOF MS, 63 (4.9%) discordant results were initially identified. Most discordant results (42/63) were due to systematic database-related taxonomical differences, 14 were explained by poor discrimination of the MALDI-TOF MS spectra obtained, and 7 were due to errors in the initial conventional identification. An extraction step was required to obtain a valid MALDI-TOF MS identification for 25.6% of the 1,278 valid isolates. In conclusion, our results show that MALDI-TOF MS is a fast and reliable technique which has the potential to replace conventional phenotypic identification for most bacterial strains routinely isolated in clinical microbiology laboratories.
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              Performance of a matrix-assisted laser desorption ionization-time-of-flight mass spectrometry system for the identification of bacterial isolates in the clinical routine laboratory.

              D Bertsch, U Betz, U M Eigner … (2008)
              Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has emerged as a new tool for the fast and reliable identification of microorganisms. We evaluated the performance of a MALDI-TOF MS-system for the identification of various clinical isolates in the routine microbiology setting. For the evaluation study a set of 1116 bacterial isolates were collected in the routine microbiology laboratory. Additonally 108 isolates of strain culture collections (ATTC, DSMZ) were utilized. Identification of the bacterial isolates was perfomed with a Microflex LT mass spectrometer in combination with the MALDI-Biotyper 2.0 software (Bruker Daltonik GmbH, Bremen, Germany). The results of the MALDI-TOF MS were compared to phenotypic bacterial identification systems used in our routine laboratory. Discrepancies were resolved by 16 S rDNA-sequencing. Of the 108 reference strains tested, 101 (93.5%) were correctly identified to species level. Overall, 1062 (95.2%) of the 1116 strains collected in the routine laboratory were correctly identified with the MALDI-Biotyper. Accuracy for the identification of Enterobacteriaceae, non-fermenting gram-negative rods, staphylococci, enterococci and streptococci with the MALDI-Biotyper was 95.5%, 79.7%, 99.5%, 100% and 93.7%, respectively. Results were available in 12 minutes for direct smear and in 20 min with an extraction method. The MALDI-TOF method proves to be a fast and reliable method for the identification of the most important bacterial isolates in the clinical laboratory.
              Article 0 comments Cited 36 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Access Microbiol
                Journal ID (iso-abbrev): Access Microbiol
                Journal ID (hwp): acmi
                Journal ID (publisher-id): acmi
                Title: Access Microbiology
                Publisher: Microbiology Society
                ISSN (Electronic): 2516-8290
                Publication date Collection: February 2021
                Publication date (Electronic): 9 December 2020
                Publication date PMC-release: 9 December 2020
                Volume: 3
                Issue: 2
                Electronic Location Identifier: 000185
                Affiliations
                [ 1​ ] departmentW. Hirsch Regional Microbiology Laboratory , Clalit Health Services , Haifa, Israel
                [ 2​ ] departmentDepartment of Evolutionary and Environmental Biology, Faculty of Natural Sciences , University of Haifa , Haifa, Israel
                [ 3​ ] departmentDepartment of Biology and Environment , Faculty of Natural Sciences, University of Haifa , Oranim, Tivon, Israel
                Author notes

                Accession numbers of the isolates: MT032343 and MT032344.

                *Correspondence: Malka Halpern, mhalpern@ 123456research.haifa.ac.il
                Author information
                https://orcid.org/0000-0002-0911-3453
                https://orcid.org/0000-0002-3407-2198
                Article
                Publisher ID: 000185
                DOI: 10.1099/acmi.0.000185
                PMC ID: 8209640
                PubMed ID: 34151142
                SO-VID: fa0678e7-a9ce-4e92-96dc-00cd44748fb6
                Copyright © © 2021 The Authors
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License. The Microbiology Society waived the open access fees for this article.

                History
                Date received : 06 September 2020
                Date accepted : 06 November 2020
                Funding
                Funded by: Israel Science Foundation
                Award ID: 2728/17
                Award Recipient : MalkaHalpern
                Categories
                Subject: Case Report
                Custom metadata
                OpenAccessEmbargo 0

                Keywords: opportunistic pathogen,tsukamurella pulmonis,acid-fast,conjunctivitis,eye infection,nasolacrimal duct obstruction
                Data availability:
                Keywords: opportunistic pathogen, tsukamurella pulmonis, acid-fast, conjunctivitis, eye infection, nasolacrimal duct obstruction

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