A system for rapidly detecting and identifying microorganisms on biological surfaces (e.g. fish skin surfaces) and / or non biological surfaces (e.g. habitat or IC surfaces) and corresponding methods.

Abstract

Abstract: The current invention comprises the novel technological development regarding the detection of fond related bacteria, such as listeria, e. coli and s. aureus in a mobile and modular design. The combination of a multiple target sensor, smart readout optics and spectral processing enables to instantly ascertain, verify and validate the presence of specific microbes. The main application is the monitoring of the fond and fond production in order to prevent the coming to the market of bacteria invested products, such as fish, chicken or other fond products destined for human consumption.

Description

Description Title: Systems, sample means and methods for rapidly detecting and identifying micro-organisms on surfaces. Field (of the invention): 5 This invention relates to systems and methods for rapidly detecting and identifying micro-organisms on biological surfaces (e.g. fish skin surfaces) and / or non-biological surfaces (e.g. habitat or IC surfaces); And sample means for these systems and methods. The invention especially finds its application in the food industry and 10 food production industry in order to prevent the coming to the market of bacteria invested products. The current invention comprises the technological development regarding the instant detection of food related bacteria. Background (of the invention): 15 Current: products for human consumption are normally tested in food production facilities through swabs and the results are being tested in laboratory facilities, which are located at a different location. The results are mostly only known after days. By that time the products for human consumption could already be consumed. 20 Publications cited: o van Dam, K. Falkena, S.A. den Daas, I. Veldhuizen. M.C.G Aalders, Improving the visualization of fingermarks using 25 multi-target immunolabeling, For Sci Int, 324, 2021 § Here a contrast agent was designed, based on immunolabeling, to bind to specific components in fingermarks. The contrast of the fingerprint was greatly enhanced, enabling better visualisation compared to 30 fingerprints enhanced with conventional methods. Comparable to the current project, we target specific components with immunolabeling. o L.S. Wilk, R.J.M. Hoveling, G.J. Edelman, H.J.J. Hardy, S. van Schouwen, H. van Venrooij, M.C.G. Aalders, Reconstructing the time since death using noninvasive thermometry and numerical analysis, Science Advances, 6(22), 2020. § An andvanced model was designed which models the energy exchange of the human body with the environment. The relevancy here is the system integration and simulation of complex processes with optimizers o M. Almasian, L.S. Wilk, P.R. Bloemen, T.G. van Leeuwen, M. ter 10 Laan, M.C.G. Aalders, Pilot feasibility study of in vivo intraoperative quantitative optical coherence tomography of human brain tissue during glioma resection, Journal of biophotonics 12 (10), e201900037, 2019. § By analysing the morphological buildup of gliomata at the 15 cellular level, we were able to create contrasts between healthy and diseased tissues which are hard to distinguish otherwise. o C.S. Stravers, E.L. Gool, T.G. van Leeuwen, M.C.G. Aalders, A. van Dam, Multiplex body fluid identification using surface 20 plasmon resonance imaging with principal component analysis, Sensors and Actuators B: Chemical, 283, 355-362, 2019. § Here a new Plasmon resonance imaging was explored to obtain the composition of several body fluids. The technique is, again based on multiple component labelling 25 o N. Achetib, L.S. Wilk, J.C.V. Schwarz, S.A.G. Lambrechts, T.G. van Leeuwen, M.C.G. Aalders, A. van Dam, Estimating the Time of Deposition of Semen Traces using Fluorescence Protein-Lipid Oxidation Signatures, Analytical chemistry, 91(5), 3204-3208, 2019. o K. Falkena, R.J.M. Hoveling, A. van Weert, S.A.G. Lambrechts, 30 T.G. van Leeuwen, M.C.G. Aalders, A. van Dam, Prediction of DNA concentration in fingermarks using autofluorescence properties, Forensic Science International, 295, 128-136, 2019. C.S. Stravers, E.L. Gool, T.G. van Leeuwen, M.C.G. Aalders, A. o van Dam, Multiplex Body Fluid Identification using Surface 35 Plasmon Resonance Imaging with Principal Component Analysis, Sensors and Actuators B: Chemical, 2018. Objects and solutions of the invention 40 The object of the invention is to overcome at least some of the above problems. This object is achieved with the system according to claim 1. The object is also achieved with a sample means according to claim 15. Furthermore the object is achieved with methods according to claims 16 and 17. Beneficial optional embodiments are the subject of the dependent claims. With the sensitive read-out device in combination with the detector / identifier means and / or presumptive read-out device according to 15 the invention, an on the spot detection and identification of micro- organisms can be achieved. This system can be easily integrated in production facilities. 20 With the systems, sample means and methods according to the invention, rapidly detecting and identifying of micro-organisms can be materialised. Especially during the production in progress in the food production facilities. The sample means according to the invention can be flexible means, such 25 that with simply swiping over all kinds of surfaces - even and uneven - micro-organisms can easily adhere to the sample means during a swiping. With the dissolvable sample means, (groups of) micro-organisms adhered 30 thereto can be easily separated from the sample means for further research. Furthermore, the dissolvement of the sample means proves the durability of the invention, since no rest product of the sample means remains. Especially with the sample means, according to the invention, with the 35 flexible usage is, e.g. a dissolvable cloth, which can easily be swiped over all kinds of even and uneven surfaces, so that micro-organisms can adhere. 40 Our invention tests the products for human consumption in-process and the results are known within minutes. Using: in-process sampling during the processing of food for human consumption. This is preferably combined with: Dissolving a nitrocellulose cloth (in case of liquid sample) Fluorescent labeling in a container Confirmatory test-high sensitivity and high selectivity Analysis and identification using machine vision algorithms High specific verification through additional tests Instant action possibilities These preferably fully computerized methods, using product-designed 10 algorithms, make the detection of harmful bacteria possible within minutes. With this method the food processing facility can undertake action immediately so as to prevent that the contaminated food will be sent out further into the food distribution process. Further benefits and advantages of the present invention will become apparent after reading the detailed description of embodiments below with appropriate reference to the accompanying figures. 20 Figure 1: The three step system described below. Figure 2: Diagram and colour changes Figure 3: Steps of the process Figure 4: several common fluorophores with their excitation (dashed line) and emission spectra (solid line) 25 Figure 5: Examples of steps 1, 2 and 3, whereby step 2 can be optional 30 Description of preferred embodiments of the invention The proposed system: Reference is made to Figure 1. 35 Hereafter the steps are described for a process for detecting Listeria and other types of bacteria on fish and meat surfaces in a processing environment with more explanation of the techniques involved, such as dissolution, fluorescent labeling, and advanced detection methods like SPRI or ring resonator detection. detecting Listeria on fish in a processing 40 environment: 1. **High throughput High sensitivity / low specificity monitoring**: remote monitoring of the product, looking for micro-organism specific autofluorescence signatures. In step 1, the conveyor belt is inspected using (1) a dedicated light source, able to emit UV and / or blue light for 5 generating the fluorescence, (2) a fluorescence-sensitive / CCD camera with filters to block the Uv / Blue excitation light and transmit the generated fluorescence light and (3) a processing unit to analyse images using machine vision algorithms, trained to recognize the deviating fluorescence patterns caused by the presence of microorganisms. We can continuously scan the product, looking for a suspicious surface and uses specific spectral signatures for determining whether a product must be further investigated (step 2) See figure 2. 15 2. **Sampling**: Use a swab or a cloth, e.g. a nitrocellulose membrane cloth, which has high affinity for biological material, sample the surfaces of the fish in the processing environment. The cloth will then contain any 20 biological material (including the microorganisms present on the surface. 3. **to a Liquid sample**: The nitrocellulose cloth will be dissolved in a solution (The overarching consequence is that the nitrocellulose is soluble in organic solvents such as acetone and esters; e.g., ethyl acetate, methyl acetate, ethyl carbonate.), This process will free the biological material, 25 including any potential Listeria bacteria, into the solvent. 4. **Fluorescent Labeling in a container (indicative test) **: Introduce fluorescent labels into the solution. These labels can be chosen based on their ability to bind to specific targets, such as Listeria bacteria. The container will be illuminated by a lightsource, with the specific 30 wavelength for the chosen fluorophore and the emission will be detected using a filtered Photodetector. If fluorescence is emitted by these labels the presence of the target bacteria is likely and for more specific analysis, the next step is necessary. 35 See figure 3. https: / / www.mdpi.com / 2079-6374 / 12 / 10 / 869 i: Fluorescent component (fluorophore, e.g.see fig 3) ii: antibodies coupled to fluorophore ->label 40 iii: solution with sampled biological material iv: reaction of target with label fluorescence when target is present. v: Fig 3: from https: / / www.mdpi.com / 2079-6374 / 12 / 10 / 869 See figure 4. 5. **Confirmatory test-high sensitivity and high selectivity **: A surface with a number of functionalized areas, being areas of immobilized antibodies for various bacterial strains and sub-strains is used. The solvent will be running over the sensor surface and specific substances bind to their intended functionalized areas. The functionalized surface can be a gold layer (in case surface plasmon resonance imaging (SPRi), the multiplex biosensor in the top figure) is used or a photonic chip (in case a Micro Ring Resonator device (MMR) is used). Both these methods can detect changes in the functionalized surface caused by the binding of the targets, indicating the presence of (in our case) Listeria bacteria. This detection can be realized within minutes. Combinations of positive areas indicate the specific strain. The device can operate in heavy processing circumstances. **Analysis and Identification**: Analyze the results of SPRi or MMR to 20 6. determine the presence and potentially the specific strain of Listeria bacteria in the sample and thus on the fish surface. This analysis may involve analysis of the SPRi-images using machine vision algorithms (these images show which area's era positive for binding and which areas are negative, see figure below. The areas with target bound (top middle and 25 left and lower right) show brighter in this image. Such combinations of bright images indicate a specific strain. Or analysis of binding kinetics from the MRR device. See: https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC3303711 / 30 See figure 5. 6. **Verification**: Because of the large impact it may be necessary to verify the presence of Listeria and potentially its substrain through 35 additional tests. 7. **Reporting and Action**: Report the findings and take appropriate action based on the results. This could involve implementing corrective measures in the processing environment to prevent contamination or ensuring proper handling of the fish products. 40 The following clauses can be defined: Claus es 1. A system for rapidly detecting and identifying micro-organisms on biological surfaces (e.g. fish skin surfaces) and / or non-biological surfaces (e.g. habitat or IC surfaces) comprising in combination: a) a presumptive detection device for determining whether a surface must be further investigated, the device comprising a dedicated light source and a fluorescence-sensitive camera for scanning a surface on which microorganisms may be present i.e. a suspicious surface, the detection of microorganisms being based on the intrinsic fluorescence (auto fluorescence) properties of the micro-organisms. b) a sample means, being means having a high affinity for biological material, and being suitable for taking a sample of biological material on 20 the suspicious surface, and e.g. being a cloth, a swab or a sponge, which sample means can collect in particular micro-organisms, c) a solvent for dissolving the sample means together with the micro- organisms collected in the sample means, d) a container (e.g.) for containing the solvent, and for containing a solution of the solvent with a dissolved sample means and dissolved micro- 25 organisms, e) a sensitive read out device for detecting and identifying micro- organisms, in particular also sub- strains thereof, comprising at least one multiple area sensor, e.g. having 48 or 96 areas, whereby specific sensor 30 means are provided on the multiple areas for binding with dissolved micro- organisms, in particular the sub-strains thereof, when a solution of the solvent and the microorganisms is running over the at least one multiple area sensor, a light source for emitting light on the at least one multiple area sensor and a detector / identifier / CCD means able to detect and identify the dissolved micro-organisms, in particular also the sub-strains 35 thereof, on the basis of the output of the at least one multiple area sensor. System according to claim 1, characterized in that 2. it further comprises reactants which are labeled with specific fluorophores and which reactants can be activated by binding to the corresponding dissolved micro-organisms, thereby changing their fluorescence characteristic, whereby the sample means comprises the reactants and / or the solvent 10 comprises the reactants, and / or the reactants are to be added to the solution in the container, the container being further suitable for containing dissolved micro- organisms binded to activated reactants, 15 the system further comprising a presumptive read out device to be used in advance of the sensitive read out device, for detecting and identifying the dissolved micro-organisms in the container, in particular main strains thereof, the device comprising a light source for exciting the fluorophores of the activated reactants and a detector / identifier / CCD means for 20 measuring the generated fluorescence and identifying the dissolved micro- organisms. 3. System according to claim 1 or 2, characterized by the sample means is a flexible cloth suitable to be swiped over the suspicious surface and being 25 made of fibers, which can comprise polyester, nylon, cellulose, chitosan, peat moss, cotton, wool, hemp, corn-based fibers. 4. System according to any of claims 1 to 3, characterized in that the cloth is a membrane or (artificial) microfiber cloth or is an artificial fabric of a nitrocellulose membrane material, which has a maximal affinity 30 for micro-organisms and is thus optimized for the adherence of microorganisms. System according to any of claims 1 to 4, characterized in that the 5. solvent is an organic solvent such as acetone or an ester; e.g., ethyl acetate, methyl acetate, ethyl carbonate. 6. The system according to any of claims 2 to 5, characterized in that the reactants are antibodies, aptamers, aptamer beacons and / or hybridized DNA displacement assays, conjugates or phages. 7. System according to any of claims 1 to 6, characterized in that the container is made of a transparent or translucent material, e.g. glass or plastic. 8. System according to any of claims 1 to 7, characterized in that the at least one multiple area sensor is a biosensor comprising 48 or 96 functionalized areas which present different reactants, especially antibodies, for reaction with different micro-organisms. 9. System according to any of claims 1 to 8, characterized in that the specific sensor means comprise functionalized gold layers which are covered by small islands of reactants, especially antibodies, the detector / identifier / CCD means making use of analyses of SPRI images (Surface Plasmon Resonance Imaging) thereby using machine vision algorithms. 10. System according to any of claims 1 to 9, characterized in that the 20 specific sensor means comprise integrated light guides on which various reactants, especially antibodies, are printed, whereby a variation in light emittance is caused by binding of microorganisms to these light guides and the analysis by the detector / identifier / CCD means is based on binding kinetics of the MMR (Mobius Micro ring Resonator). 25 11. System according to any of claims 1 to 10, characterized by the dedicated light source of the presumptive device is a light source able to emit UV and / or blue light and the camera of the presumptive detection device / CCD is a hyper spectral camera (pushbroom configuration). 12. System according to any of claims 1 to 10, characterized by the 30 presumptive device further comprising a dedicated light source able to emit UV and / or blue light for generating the fluorescence, the fluorescence-sensitive camera comprises filters to block the UV / blue excitation light and transmits the generated fluorescence light and a processing unit to analyse images using machine vision algorithms, trained 35 to recognize the deviating fluorescence patterns caused by the presence of microorganisms, whereby this combination is able to continuously scan a suspicious surface and uses specific spectral signatures for determining whether a surface must be further investigated. 13. The system according to any of claims 1 to 12, characterized in that it further comprises a conveyor belt for conveying products in a product processing facility, such as fish in a fish processing facility whereby the fish have a skin surface. 14. The system according to any of claims 1 to 13, characterized in that the microorganisms are bacteria such as MRSA, e- coli or listeria. 15. Sample means as defined in any of claims 1-4 and 6 in combination with a solvent as defined in any of the claims 1, 2, 5 and 6 OR solution of such sample means dissolved in such a solvent, as a set or part of a set for sampling. 16. Method for rapidly detecting and identifying microorganisms on biological surfaces (e.g. fish skin surfaces) and / or non-biological surfaces (e.g. habitat or IC surfaces) using the system according to claims 1 and 2 and optionally any of claims 3- 15, comprising the steps of a. scanning with a presumptive detecting device a suspicious surface, 20 detecting and identifying microorganisms based on the intrinsic fluorescence (auto fluorescence) properties of the microorganisms, deciding on the basis hereof to take a sample in order to further investigate the surface, b. in case of further investigation, 25 using sample means to take a sample of material present on the surface comprising micro-organisms, e.g. by swiping the sample means over the surface thereby adhering or absorbing micro-organisms to the sample means, c. providing a container with a solvent, d. making a solution in the container by dissolving the sample means 30 together with the micro-organisms in the solvent, e. adding reactants labeled with specific fluorophores to the solvent or to the sample means before making the solution, or adding such reactants to the solution, f. using a presumptive read out device for detecting and identifying the micro-organisms, in particular the main strains thereof, and deciding on the basis hereof to continue with further investigation of the micro-organisms, in case of further investigation of the micro-organisms, g. supplying the solution in the container to a sensitive read out device, and detecting / identifying the respective specific micro-organisms, in 10 particular also the sub-strains thereof, which are present on the surface. 17. Method for rapidly detecting and identifying microorganisms on biological surfaces (e.g. fish skin surfaces) and / or non-biological surfaces (e.g. habitat or IC surfaces) using the system according to any of 15 claims 1- 15, comprising the steps of scanning with a presumptive detecting device a suspicious surface, a. detecting and identifying microorganisms based on the intrinsic fluorescence (auto fluorescence) properties of the microorganisms, 20 deciding on the basis hereof to take a sample in order to further investigate the surface, b. in case of further investigation, using sample means to take a sample of material present on the surface comprising micro-organisms, e.g. by swiping the sample means over the suspicious surface thereby adhering or absorbing microorganisms to the 25 sample means, c. providing a container with a solvent, d. making a solution in the container by dissolving the sample means together with the microorganisms in the solvent, e. supplying the solution in the container to a sensitive read out device, and detecting / identifying the respective specific microorganisms, in particular also the sub-strains thereof, which are present on the surface. 5

Claims

1. A system for the rapid detection and identification of micro- organisms on biological surfaces (for example, fish skin surfaces) and / or non-biological surfaces (for example habitat or IC) surfaces), comprising in combination: a) A 'first detection device' for determining whether a surface is examined must be, the device contains a dedicated light source and a fluorescent camera for scanning a surface on which microorganisms may be present, that is to say a suspect surface, the detection of microorganisms being based on the intrinsic fluorescence (autofluorescence) properties of the microorganisms. b) Sampling agents, where the agents have a high affinity for the biological material, and suitable for taking a sample of the biological material on the suspect surface, and for example, being for example a cloth, cotton swab or a sponge, which in particular in can collect specific microorganisms. c) A solvent for dissolving the sample media together with the microorganisms collected in / with the sample. A container (for example) that holds the solution, and for the d) containing (a) solvent(s) of the solvent with a dissolved 20 sample and the dissolved microorganisms A sensitive readout device for the detection and identification of e) microorganisms, particularly regarding their substrains, comprising at least one multiple area sensor, for example having 48 or 96 areas, where specific sensor devices have been applied to the 25 multiple regions for binding with the dissolved microorganisms, especially the sub-strains thereof, when a solution of the solvent and the microorganisms over at least one multiple area sensor, a light source for shining light on at least one multiple area sensor and a detection device / identification device capable 30 being to detect and identify the dissolved microorganisms, especially regarding the sub-strains thereof, based on the output of at least one of the multiple area sensors. 35 2. System according to claim 1, characterized by that it further includes reactants that are labeled with specific fluorophores and which reactants can be activated by binding the corresponding dissolved microorganisms, hereby their fluorescent characteristics changing, where the sampling means comprise the reactants and / or the solvent the includes reactants, and / or the reactants must be added to the solution in the container, The container is also suitable for containing dissolved micro- organisms linked to activated reactants, The system further includes a suspected reading device, to be determined in advance. use prior to the sensitive readout device, for detection and the identification of the dissolved microorganisms in the container, in particular where the main stems thereof are concerned, the apparatus containing a light source for stimulating the fluorophores of the activated reactants and a detector / identification device / CCD for measuring the generated fluorescence and the identification of the dissolved micro- organisms 3. System according to claim 1 or 2, characterized by the sample, is a flexible cloth suitable for wiping over the suspected surface, 20 consisting of fibers, which may contain: polyester, nylon, cellulose, chitosan, peat moss, cotton, wool, hemp, corn-based fibers.

4. System according to one of claims 1 to 3, characterized by the fact that the cloth is a membrane or is an (artificial) microfiber cloth or a artificial fabrication of a nitrocellulose membrane material is, that a 25 has maximum affinity for microorganisms and is therefore optimized for adhesion to microorganisms.

5. System according to one of claims 1 through 4, characterized by the fact that the solution is an organic solution, such as acetone or an ester; for example ethyl acetate, methyl acetate, ethyl carbonate. 30 6. The system according to one of claims 2 through 5, characterized by the fact that the reactants are: antibodies, aptamers, aptamer beacons and / or hybridized DNA displacement assays, conjugates or phages.

7. The system according to one of claims 1 through 6, characterized by the fact that the container consists of a transparent or translucent material, 35 for example glass or plastic.

8. The system according to one of claims 1 through 7, characterized by the fact that at least one multiple area sensor is a biosensor, containing 48 or 96 functionalized regions containing different reactants present, specifically antibodies, for reaction with the various micro- organisms The system according to one of claims 1 through 8, characterized by the 9. fact that the specific sensor media comprise functionalized gold layers, which are covered with reactant islands, specifically antibodies, the detection method / the identifier / CCD makes use of analyses of SPRI images (Surface Plasmon Resonance Imaging), using algorithms driven by machine image processing.

10. The system according to one of claims 1 through 9, characterized by the fact that the specific sensor devices contain integrated light guides on which various reactants, particularly antibodies, are placed, where a variation in light emissions is caused by the binding of the microorganisms on these light guides and the analysis by the detection device / identification device / CCD is based on connecting kinetics of the MMR (Mobius Micro ring Resonator).

11. The system according to one of claims 1 through 10, characterized by 20 the fact that the specific light source of the device that the suspected demonstrates presence, is a light source capable of UV light and / or to emit blue light and the camera of the device that the suspected demonstrates presence of a hyperspectral camera (pushbroom configuration / CCD).

12. The system according to one of claims 1 through 10, characterized by 25 the fact that the device demonstrating the suspected presence, further includes a specific light source capable of emitting UV light and / or blue light to push to create the fluorescence, the fluorescence-sensitive The camera includes filters to the UV excitation light / blue excitation light blocking and the transmission of the generated fluorescent light and a 30 processing device for analyzing images, using algorithms driven by machine image processing to determine whether a surface needs to be investigated further.

13. The system according to one of claims 1 through 12, characterized by the fact that it further includes a conveyor belt for transporting 35 products in a food processing facility, such as fish in a fish processing site where the fish have a skin surface, but also such as chicken and other meat products in a chicken and meat processing plant, where the chicken and / or the meat product has a skin surface. The system according to one of claims 1 through 13, characterized by 14. the fact that the microorganisms are bacteria such as MRSA, E. coli or Listeria. Sampling agents as defined in any of claims 1, 4 and 6 in 15. combination with a solvent as defined in one of the claims 1, 2, 5 and 6 OR solution of such sampling agents dissolved in such a solvent, as a set or part of a set for sampling.

16. Method for the rapid detection and identification of microorganisms on biological surfaces (for example fish skin surfaces) and / or non- biological surfaces (for example, habitat or IC surfaces) using the system according to claims 1 and 2 and optionally one of claims 3 - 15, consisting of the steps a. scanning a suspicious surface with the first detection device for determining whether a surface needs to be examined, detecting and identifying microorganisms based on intrinsic fluorescence (autofluorescence) properties of the microorganisms, based on this decide to take a sample, with the aim of the 20 surface to investigate further, b. in the event of further investigation, use sampling equipment to take a sample of the material that is present on the surface which contains microorganisms, for example by the wiping of the agent with the aim of obtaining the sample, whereby the 25 microorganism attaches to or is absorbed by the sample collection, making a container (for example) with a solvent available, c. d. creating a solution in the container by means of solving of (a) sampling medium(s) together with the microorganism in the 30 solvent, e. adding reactants labeled with specific fluorophores to the solvent or to the sample before the solvent was made becomes, or the addition of such reactants to the solution, f. make use of the first detection device for detecting and identifying microorganisms, in particular the main strains thereof, and decide based on this whether to continue research into microorganisms, When the microorganisms are further examined, g. Adding the solution in the container to a sensitive reading system, that Detects / identifies the respective specific microorganism (CCD), especially its sub-stems, which are present on the surface.

17. Method for the rapid detection and identification of microorganisms on biological surfaces (for example fish skin, chicken and animal skin) surfaces) and / or non-biological surfaces (for example, habitat) or IC surfaces) using the system according to one of the claims 1 - 15, comprising the steps a. Scanning a suspicious surface with the initial detection device to determine whether a surface needs to be examined, detect and identify microorganisms based on the intrinsic 20 fluorescence (autofluorescence) properties of microorganisms, based on this, determine to take the sample with the aim of to investigate surface further, b. in the event of further investigation, to use the sampling equipment to take a sample of the present 25 material on the surface, consisting of microorganisms, for example by wiping a sampling agent over the suspect surface, whereby microorganisms adhere to the sampling medium or are affected by the sampling medium be absorbed, making a container available, for example, with a solvent, c. 30 d. making a solution in the container by dissolving the sample medium together with the microorganisms in the solvent, e. Providing the solution in the container to a sensitive reading device, and detect / identify the respective specific micro- organisms, in particular also the sub-phyla thereof, which are present 5 on the surface. The proposed system: Confirmatory test Indicative test Light source 1111 Detector « Color indicates presence of Buffer dissolvable absorbent tissue hazardous substances against Step 2c: Step 2a: Step 2b: Read out of chemical Apply tissue to item of Transfer tissue to reaction buffer Step 1: Apply specific light source to reaction based on color interest Step 3: Multiplex assay detect deviations in (fluorescent) spectra change Figure 1