Microbial Analysis and Inactivation Strategies

The current bioburden caused by bacterial pathogens in the areas of human medicine and veterinary medicine, and at the transmission from animals to humans, and not least the multi-resistant pathogens now emerging, make new and effective measures necessary for both combating and diagnosing these germs.

Highly specific antigens from the corresponding species of pathogen, isolated from an infection-relevant transcriptome, provide a basis for this. Depending on the situation, this involves isolating mRNAs from the pathogens using microarrays, and characterizing their resulting expressions for immunological purposes. Only the genes which are actually active when an infection occurs or their transcription products in the form of mRNA are therefore used as a basis for the identification of antigens. The method is fast and effective, and only requires a few cleaning steps.

Furthermore, the developments allow new instruments for immunodiagnostics (for example, highly specific antibodies) and new potential for generating selective vaccines to be derived. The method can be applied to any germ, and has already been used for a range of different pathogens, such as salmonella, campylobacter, staphylococcus, klebsiella, neisseria, pseudomonas, streptococcus and clostridium.

Development of an Apparatus-Free, Universal Sensor for Direct Detection of Bacteria – Apparatus-Free Bacteria Sensor

The cooperative project describes a specific, simple and fast means of detecting bacterial contamination of water, giving the project high relevance for the master plans pursued by the Berlin-Brandenburg Food Industry and Health Care Clusters in the region, thereby exploiting synergies with these two clusters. By employing a new method established at the Fraunhofer IZI-BB which is used to obtain antigens from bacterial mRNA, monoclonal antibodies are produced that are reduced to Fab fragments and coupled with an electron transfer resonance element that can be detected optically. A specific analysis architecture based on nanobeads allows a visible color signal to be produced when bacterial pathogens are present, which highlights the specific germs.

Logo-EFRE-Brandenburg

This project is being funded by ERDF.
The Fraunhofer IZI-BB would like to thank the funding authority for the opportunity to realize this research project.

Methods

  • Prokaryotic cDNA expression libraries
  • Phage display
  • Microbial knock-out mutants

 

Equipment

  • Anaerobic glovebox

  • RIPAC-Labor GmbH
  • Bovicare GmbH
  • ILBC GmbH
  • Robert Koch Institute
  • IMTEK Freiburg
  • FU Berlin, IMT

  • Connor DO, Danckert L, Hoppe S, Bier FF, von Nickisch-Rosenegk M. Epitope
  • determination of immunogenic proteins of Neisseria gonorrhoeae. PLoS One. 2017
  • Jul 19;12(7):e0180962. doi: 10.1371/journal.pone.0180962. eCollection 2017.
  • PubMed PMID: 28723967; PubMed Central PMCID: PMC5516995.
  • Connor DO, Zantow J, Hust M, Bier FF, von Nickisch-Rosenegk M. Identification
  • of Novel Immunogenic Proteins of Neisseria gonorrhoeae by Phage Display. PLoS
  • One. 2016 Feb 9;11(2):e0148986. doi: 10.1371/journal.pone.0148986. eCollection
  • 2016. PubMed PMID: 26859666; PubMed Central PMCID: PMC4747489.
  • Danckert L, Hoppe S, Bier FF, von Nickisch-Rosenegk M. Rapid identification of novel antigens of Salmonella Enteritidis by microarray-based immunoscreening. Microchimica Acta 02/2014; 3.43 Impact Factor
  • Hoppe S, Bier FF, von Nickisch-Rosenegk M. Rapid Identification of Novel Immunodominant Proteins and Characterization of a Specific Linear Epitope of Campylobacter jejuni. PLoS ONE 01/2013; 8(5):e65837.
  • Hoppe S, Bier FF, von Nickisch-Rosenegk M. Microarray-based method for screening of immunogenic proteins from bacteria. Journal of Nanobiotechnology 03/2012; 10:12.