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US-12618756-B2 - Device containing glass beads functionalized with polyethyleneimine, and use thereof for capturing microorganisms

US12618756B2US 12618756 B2US12618756 B2US 12618756B2US-12618756-B2

Abstract

A device containing glass beads functionalized with polyethyleneimine which is adsorbed on the surface of the glass beads, and the use thereof for capturing microorganisms for implementing a method for removing microorganisms or for diagnosing. The microorganisms can be chosen in particular from bacteria and Fungi, in particular yeasts and fungi.

Inventors

  • Wilfried ABLAIN

Assignees

  • MICROBS SAS

Dates

Publication Date
20260505
Application Date
20201027
Priority Date
20191031

Claims (13)

  1. 1 . A method for capturing living microorganisms comprising a step of bringing into contact, a liquid or viscous sample containing the microorganism, with a device comprising a hollow container containing non-magnetic glass beads consisting of a soda-lime glass or a borosilicate glass with polyethyleneimine adsorbed on the surface of the glass beads, wherein the molecular mass of the adsorbed polyethyleneimine is comprised of 0.6 to 2000 kDa, under conditions making it possible to create an interaction between the microorganism and the glass beads, and to obtain the microorganism captured on the glass beads, wherein a proportion of the microorganisms originating from the sample and captured on the glass beads is between 0.001% and 100% for a debacterization of said sample.
  2. 2 . The method according to claim 1 , comprising an additional step of eluting the previously captured microorganisms under conditions allowing the separation of the microorganisms captured from the glass beads and the recovery of the microorganism.
  3. 3 . The method according to claim 1 , wherein the microorganisms are selected from the group consisting of bacteria and fungi, and, wherein said fungi is selected from the group consisting of: Absidia, Alternaria, Aspergillus, Aureobasidium, Botrytis, Brettanomyces, Byssochlamys, Candida, Chaetomium, Cladosporium, Colletotrichum, Cryptococcus, Debaryomyces, Emericella, Epicoccum, Eupenicillium, Eurotium, Fusarium, Galactomyces, Geotrichum, Gliocladium, Hanseniaspora, Humicola, Hyphopichia, Kluyveromyces, Lichtheimia, Lodderomyces, Meyerozyma, Monascus, Mucor, Mycocladus, Neosartorya, Nigrospora, Paecilomyces, Penicillium, Pestalotia, Phoma, Phytophthora, Pichia, Pythium, Rhizoctonia, Rhizopus, Rhodotorula, Saccharomyces, Saccharomycopsis, Schizosaccharomyces, Sclerotinia, Scopulariopsis, Serpula, Stemphylium Talaromyces, Thielaviopsis, Torulaspora, Trichoderma, Trichosporon , Trichothetium, Ulocladium, Verticillium, Wallemia, Wickerhamomyces, Xylaria , and Zygosaccharomyces.
  4. 4 . The method according to claim 3 , wherein the microorganisms are bacteria, and said bacteria is selected from the group consisting of Acetobacter, Achromobacter, Acidovorax, Acinetobacter, Actinomyces, Aerococcus, Aeromonas, Alcaligenes, Alicyclobacillus, Aquaspirillum, Asaia, Bacillus, Bifidobacterium sp., Bordetella, Brachybacterium, Brevibacillus, Brevibacterium, Brevundimonas, Burkholderia, Buttiauxella, Campylobacter, Carnobacterium , Cellulomona, Citrobacter, Clavibacter Clostridium, Corynebacterium , Cronobacte, Cupriavidu, Curtobacterium, Elizabethkingia , Enteractinococcus, Enterobacter, Enterococcus, Escherichia, Flacklamia, Flavobacterium, Geobacillus , Glutamicibacte, Halobacillus, Klebsiella, Kocuria, Lactobacillus, Lactococcus, Leclercia , Lelliottia, Leuconostoc, Lysinibacillus, Macrococcus , Methylobacteriu, Microbacterium spp. (CDC A-5), Micrococcus, Moraxell, Mycobacterium, Nesterenkonia, Oceanobacillus sp, Ochrobactrum, Paenibacillus, Pandorae, Pantoea, Paracoccus, Pasteurell, Pediococcus, Propionibacterium, Proteus, Pseudomonas, Ralstonia, Rhizobium, Roseomona, Rothia, Salmonella, Sanguibacter, Serratia, Shewanella, Sphingomonas, Sporolactobacillus, Sporosarcina, Staphylococcus, Stenotrophomonas, Streptococcus, Streptomyces, Thermoanaerobacterium, Variovorax , and Virgibacillus.
  5. 5 . The method according to claim 1 , wherein the liquid or viscous sample is chosen from the group consisting of: a biological, human or veterinary sample, a pharmaceutical sample, a cosmetic sample, a food sample, a sample of process water, and an environmental type sample.
  6. 6 . The method, according to claim 5 , wherein the liquid or viscous sample is a biological, human or veterinary sample is selected from the group consisting of: urine, blood, synovial fluid, lymph, liquid lachrymal, secretions and mucous membranes.
  7. 7 . The method, according to claim 5 , wherein the liquid or viscous sample is a pharmaceutical sample selected from the group consisting of: solutions for injection, syrups, vaccines, eye drops and ophthalmic gels.
  8. 8 . The method, according to claim 5 , wherein the liquid or viscous sample is a cosmetic sample selected from the group consisting of: make-up removers, products for cleaning the skin, deodorants, products intended for shaving, self-tanners, sun protection creams, solvents, shampoos and conditioners.
  9. 9 . The method, according to claim 5 , wherein the liquid or viscous sample is a sample of process water that is a sample from the loops of cleaning.
  10. 10 . The method, according to claim 5 , wherein the liquid or viscous sample is an environmental type sample selected from the group consisting of: marine and rain samples.
  11. 11 . The method, according to claim 5 , wherein the liquid or viscous sample is a food sample selected from the group consisting of: drinks, meats, ready meals, dairy products and egg products.
  12. 12 . The method, according to claim 11 , wherein the liquid or viscous sample is a drink selected from the group consisting of: water, milk, fruit juices, sodas, alcoholic beverages and tea-based beverages.
  13. 13 . The method, according to claim 12 , wherein the liquid or viscous sample is orange juice.

Description

FIELD The present invention relates to a device containing functionalized glass beads, and its use in capturing micro-organisms for the implementation of a method for the elimination of microorganisms or for diagnosis. The capture of microorganisms is of fundamental interest in many types of industry, such as the pharmaceutical, cosmetic, veterinary or food industries. It can in particular be used for applications which can be divided into two main areas: The elimination of micro-organisms from potentially contaminated solutions,Analysis in the form of a diagnosis, in particular in the context of a clinical diagnosis, making it possible to assess the microbiological quality of a solution. BACKGROUND To eliminate microorganisms present in potentially contaminated solutions, the techniques used generally involve heat treatment (Magali, WAGNER, Anne Gaëlle MELLOUET, and François ZUBER. 2016. “Continuous heat treatment of pumpable products.” “Agribusiness. “Engineering Techniques, Sep. 10, 2016) and/or membrane filtration (Christel, CAUSSERAND, Claire ALBASI, and Hélenè ROUX DE BALMANN. 2017. “Membrane filtration (RO, NF, UF, MF)—Applications in water treatment.” “Water technologies.” Engineering Techniques, Aug. 10, 2017). The major drawback of heat treatment comes from the fact that the temperature is likely to cause irreversible changes to the product by acting directly on its constituents. Also, at the microbiological level, the heat treatment corresponds to a reduction in the load of microorganisms and it is for example likely to reactivate bacterial spores. Regarding membrane filtration, clogging of the membrane is the main problem that can be encountered. The latter may be due to the concentration of microorganisms in the product as much as to the nature of the product and in particular to the solid particles present within it. Microbiological analysis requires the use of precise techniques for which the time to obtain the shortest possible result is sought. Indeed, the faster the analysis results are obtained, the more it is possible to initiate corrective actions in the event of unsatisfactory or unacceptable results. In particular, in the medical field, it is necessary to predict and diagnose the risk of infection: the faster and more accurate the diagnosis, the more effective the care of patients and the minimized risk of transmission. However, to demonstrate the presence of microorganisms, it is necessary to take sufficiently large samples to ensure that a minimum quantity of microorganisms is recovered. It is then necessary to increase their concentration, isolate them and identify them. A key step for most of these methods for detecting/quantifying microorganisms in samples that are liquid or made liquid, is their ability to exceed the detection limit of the evaluation technique used. This is particularly difficult for samples with a low concentration of microorganisms of interest. An enrichment or concentration step often appears necessary for this type of sample, said enrichment step possibly being implemented directly in the packaging in which the sample is located. In this case, it is an enrichment step directly in the product. The enrichment step requires the use of culture media, selective or not, which aim to promote the growth of target microorganisms in biological or environmental samples, while limiting the growth of non-target flora. Thus, the target population, which is often present at low levels compared to the appendix flora present in food, is amplified. These enrichment steps upstream of the analysis make it possible to increase the number of microorganisms of interest in the sample but condemn both the possibility of a rapid analysis but also the possibility of quantifying the initial population of microorganisms of interest. The other possibility is to go through a stage of concentration of the microorganisms from the liquid sample or made liquid. One of the most widely used techniques for the concentration of microorganisms from samples is the use of one or more membrane filters of variable porosity through which the liquid medium is filtered. The microorganisms contained in the sample are stopped by the membrane and therefore concentrated. Such a technique is generally implemented for the microbiological analysis of water, drinking water, beverages, or pharmaceutical products. Although easy to use, this membrane filtration method remains limited by factors causing clogging of the membrane filter such as high turbidity, or the presence of particles in the sample. Other factors related to the nature of the filter and its sterilization method can also influence the result of the microbiological analysis, which can seriously alter the viability, precision and sensitivity of the method and lead to erroneous and poorly reproducible results. We can cite, by way of non-exhaustive example, the inhibition of microbial growth, an abnormal propagation of colonies, the existence of non-wet