Molecular detection of microorganisms
We use molecular imaging and other molecular methods to detect microorganisms. By combining fluorescence in situ hybridization (FISH) and 16S rRNA gene sequencing, microorganisms can be identified and visualized in clinical specimens.
Our methods
We combine molecular imaging with molecular microbiology
- FISHseq: fluorescence in situ hybridization (FISH) + 16S rRNA gene sequencing
- MG-FISH®: fluorescence in situ hybridization (FISH) + Next Generation Sequencing (NGS, microbiome analysis)
- quantitative Real-time PCR (qPCR)
FISHseq (FISH + 16S rRNA gene sequencing)
FISH is a microscopic method, that combines the advantages of molecular biology, histology and microscopy. FISH uses fluorescence labeled probes, which bind complementary to specific regions of microbial rRNA. Depending on the used probe, bacteria and fungi can be detected on a domain, genus or species level. FISH can quantify their activity on a single cell level based on ribosome content. Furthermore, FISH visualizes microorganisms in situ, therefore the identity, amount, localization and spatial formation can be determined.
For sequencing, we amplify part of the 16S rRNA gene by PCR and use Sanger sequencing to determine the identity of microorganisms
Further information:
Quality Control in Diagnostic Fluorescence In Situ Hybridization (FISH) in Microbiology.
Kikhney J, Moter A. Methods Mol Biol. 2021;2246:301-316. doi: 10.1007/978-1-0716-1115-9_20.
Fluorescence in situ hybridization (FISH) in the microbiological diagnostic routine laboratory: a review.
Frickmann H, Zautner AE, Moter A, Kikhney J, Hagen RM, Stender H, Poppert S. Crit Rev Microbiol. 2017 May;43(3):263-293. doi: 10.3109/1040841X.2016.1169990. Epub 2017 Jan 27.
MG-FISH® (FISH + Next Generation Sequencing (NGS))
In contrast to Sanger sequencing, which is used in FISHseq, microbiome analysis by Next generation sequencing (NGS) can analyze all microorganisms in a specimen simultaneously. MG-FISH® is especially useful in specimens, which contain multiple species, and can visualize multispecies biofilms.