AnagnosTec SARAMIS™ - Spectral ARchive And Microbial Identifications System
Rapid and reliable Identification of mircoorganisms without extensive manipulation is the major interest at microbiology. The SARAMIS™ software and database of human and veterinary pathogens, food pathogens, environmental and natural product producing microorganims has been developed using MALDI-TOF (Matrix Assisted Laser Desorption Ionization-Time-of-Flight) mass spectrometry.
The modular microorganisms identification system AnagnosTec SARAMIS™ permits the archiving and the evaluation of MALDI-TOF MS data in three different operating modes: reflector, PSD and linear (“fingerprint”).
AnagnosTec SARAMIS™ includes a patented procedure for statistic evaluation and weighting of data. This procedure allows the identification of microorganisms. In addition, theoretical spectra (SuperSpectra™), which can be obtained using a verified and patented procedure, help to multiply the efficiency of the microbial identification and the grouping of samples.
The implementation of universal interfaces makes SARAMIS™ compatible with mass spectrometers of various manufacturers. MS data are transformed into a universal highly compressed format which reduces the data volume by 99% and allows fast data retrieval and evaluation.
Every strain produce a MALDI-TOF mass fingerprint pattern with specific signals at strain, species, genera and family level. These specific signals can be used as mass pattern of biomarkers in superspectra for automated identification of unknown strains.
Requirements for modern methods suitable for microorganism identification and characterization are robustness, simple handling, low costs, speed and high-throughput capability. Therefore, mass spectrometry may offer significant advantages over classical technologies e.g., polymerase chain reaction, sequencing, electrophoretic separation of nucleic acid fragments. The detection limit for the MALDI-TOF MS analysis was determinate below 100 cells.
The detected peak pattern were compared and stored in SARAMIS™ database. The identification of unknown strains was performed using the supserspectra tool. These superspectra are generated based on the measurement of known microorganisms. Thereby, a reference data base is generated which is continuously growing by feeding in new spectra and species. The SuperSpectra™ contain only peaks which are informative for the respective group of microorganisms. Classifacation by comparison with the reference spectra database is possible down to the strain level. These results were equal or even superior compared to traditional methods while the procedure is much faster, up to one hundred measurements per hour, with minimal consumables costs.
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Key features are:
- User-friendly Data compression, search and storage
- Visualisation of MS data
- Archiving in individually user defined adaptable family tree structures
- Identification of microorganisms using internal reference databases of spectra
- Taxonomic grouping of strains like dendrogramms
- Comparison of spectra of unknown microbial isolates and identification of redundant samples
- User-friendly search engine (e.g. search by mass, file number, organism, user, text etc.)
- Support for automated spectra comparison over selected database modules, mass regions and intensity values
- SuperSpectrum – a theoretical spectrum collected from the measured spectra of certain genus, species, type and strains. With the Superspectra it is possible to identify the microorganisms correctly and automatically.
Application and Conclusion
- Useable for all microorganisms like bacteria, yeast and fungi
- Rapid identification of microorganisms/isolates in clinical, food and animal microbiology
- Rapid classification of environmental microorganisms like dendrogramm
- Taxonomic clustering of new or modify strains (e.g. mutants)
- Identification of S3 and S4-microorganisms (
specific inactivation method from Robert Koch Institute Berlin) - Dereplication of new isolates/strains
- Identification of mixture sample using Superspectra tool
- Very easy and rapid sample preparation (1 minute)
- Minimal sample preparation (single cell colony)
- Different culture media and conditions did have influence for identification
- detection limit below 100 cells
- Fast and automated MALDI-TOF measurement
- High reproducibility from different growth media, conditions or growing and cell state
- No preselection of growth conditions, reagents, consumables ore reactions
- Low cost identification of all microorganisms
- Detection of Susceptibility testing is possible
- Direct detection of toxins is possible
- Additional and parallel analysis of natural products, lipids or other cell metabolites
- Past of data from published, theoretical and other spectra are possible
- Usable for all simple linear MALDI-TOF MS instruments
Sample Preparation
The Bacteria, yeast and fungi can be growth on standard Agar plates over night, or as flask culture. All standard media can use for growing the microorganisms. For the MALDI target sample preparation, please select a single cell colony (small amount of cells) and transfer to the MALDI target. Fresh material or dried cells are useable. The cells can be transferred directly on the target and immobilised immediately after addition of 1 µl Matrix (10 mg/ml 2,5-dihydroxybenzoic acid in water/acetonitrile (1:1) with 0.03% trifluoroacetic acid). The analysis can be performed using a simple linear MALDI-TOF mass spectrometer with nitrogen laser (337 nm). The data acquisition range is from m/z = 2.000 to 20.000. After smoothing, baseline correction and peak detection steps can be transfer the peak lists directly to the SARAMIS-software. In the SARAMIS-software can be analysed the pattern with comparison function and identify the unknown isolates.