ASSESSMENT OF THE HUMAN MICROECOLOGICAL STATUS BY CHROMATOGRAPHY-MASS SPECTROMETRY
Patent: https://patents.google.com/patent/RU2501011C2/en
Indications for use of the medical technology
- Determination of the microecological status of the organism and its deviations from homeostasis.
- Identification or clarification of etiology of the infectious and inflammatory process in any nosological entities in clinical practice.
Characteristics of the method
- Determination of more than 50 microorganisms simultaneously in one single test
- Versatile, works for different groups of microorganisms: bacteria, fungi, viruses
- Test results available in 3 hours
- Sensitivity 104-105cells per sample
- Selectivity – up to species in the presence of a marker
- Analysis directly in the material without the need to plate and cultivate
- Does not require biological and biochemical test materials – culture media, enzymes, primers, etc.
Essence of the method
The method relates to a new direction in microbiological research – diagnosis of infections, dysbiosis and inflammatory processes by specific microbial chemicals (markers). These substances are contained in the cell walls of microorganisms or produced by them during their vital activity.
The diagnosis by chemical markers of microbial cells is possible because their chemical structure differs from the substance of human cells. In this case, we are talking about various fatty acids: humans have slightly over 20 species, while microbes – over 200.
Therefore, it is not difficult to detect the presence of microbes in the human body, if there is a sensitive enough method of analysis. The chromatography-mass spectrometry is such a method (more details: Wikipedia – Gas chromatography–mass spectrometry).
GC-MS is a combination of gas chromatography, a separation technique, and mass spectrometry, which is used to identify analytes. This detection technique has extensive applications in biomedicine, including disease diagnosis, pharmaceutical quality control, and metabolic profiling.
GC-MS can be used for the bioanalysis of body fluids to detect narcotics, barbiturates, alcohols, and drugs. It is also useful in detecting pollutants and metabolites in serum and in fatty acid profiling in microbes. GC-MS has specific advantages that led to labeling this technology as the “gold standard” in
metabolomics (Lu et al, 2008). This is due to the robust, reproducible and selective nature of the technique, as well as the large number of well-established libraries and metabolite databases available (NIST Mass Spectral Library; MIDI Sherlock).
Being supported by cutting-edge software and analysis methodologies, these methods make it possible to quickly and reliably determine small proportions of substances of microbial origin in any human biological fluids and in environmental objects.
Within a few hours, it is possible to qualitatively and quantitatively determine the composition of any microorganisms, if they have markers or differ in the chemical component profile, just the way people differ in fingerprints.
Chromatography vs. Genetic-based methods
The method of detecting microorganisms by FA-markers is similar to genetic analysis (PCR, 16s rRNA nucleotide sequence determination, etc.), since the composition of fatty acids is determined in DNA and is reproduced by replicating a portion of the genome by transport RNA and subsequent synthesis of FA in mitochondria by matrix RNA.
Therefore, the fatty acid profile of bacteria is their unique profile ID, like people’s fingerprints. It is as conservative as the structure of DNA, but also susceptible to mutations under the influence of environmental factors.
The stability of the set of fatty acids that make up the cells of microbes is confirmed by studies in bacterial paleontology, which show that, if we look back in time up to 2.5 billion years, the composition of the fatty acids of individual microbes and the pool of their fatty acids as a whole remains constant.
Scientific fundamentals and key contributors:
This diagnostic method is based on numerous studies of recent decades in the field of fatty acid composition of microbial cells and chemo differentiation of microorganisms and is based on the works of famous scientists:
G.A. Osipov, Bakulev Scientific Center for Cardiovascular Surgery, Moscow, Russia – Read More
D.C.White, University of Tennessee, Knoxville, USA – Read More
Lennart Larsson, Division of Medical Microbiology, Lund University, Lund, Sweden – Read More
Jeremy K. Nicholson, Section of Biomolecular Medicine, Imperial College London, UK – Read More
B. Brooks, Center for Disease Control, Atlanta, USA – Read More
Beloborodova (Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Moscow, Russia)
P.D. Nichols (Research Scientist, CSIRO, Canberra, Australia)
C.W. Moss (Case Western Reserve University, Cleveland, USA)
Yanzen (GALAB Laboratories GmbH, Hamburg, Germany)
B. Drucker (Department of Bacteriology and Virology, University of Manchester, Lancashire, UK)
Asselineau (Institute of Pharmacology and Structural Biology, CNRS-Paul Sabatier University research center, Toulouse, France)
Goodfellow (Newcastle University, Newcastle, United Kingdom)
D.E. Minnikin (Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, UK)