WO2008091851A1 - Procédé et dispositif d'analyse rapide permettant d'estimer la contamination microbienne de substrats différents - Google Patents

Procédé et dispositif d'analyse rapide permettant d'estimer la contamination microbienne de substrats différents Download PDF

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Publication number
WO2008091851A1
WO2008091851A1 PCT/US2008/051631 US2008051631W WO2008091851A1 WO 2008091851 A1 WO2008091851 A1 WO 2008091851A1 US 2008051631 W US2008051631 W US 2008051631W WO 2008091851 A1 WO2008091851 A1 WO 2008091851A1
Authority
WO
WIPO (PCT)
Prior art keywords
collection device
receptacle
media
microbial contamination
tape
Prior art date
Application number
PCT/US2008/051631
Other languages
English (en)
Inventor
Vladimir Gouli
Rosanna Giordano
Svetlana Gouli
Original Assignee
University Of Vermont Of Burlington, Vt
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University Of Vermont Of Burlington, Vt filed Critical University Of Vermont Of Burlington, Vt
Publication of WO2008091851A1 publication Critical patent/WO2008091851A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/10—Petri dish
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements

Definitions

  • the present invention relates to a rapid method to estimate microbial contamination from different substrata, such as solid, soft or gelatinous materials, and more particularly, a device for rapidly detecting and identifying both aerobic and anaerobic microorganisms.
  • One aspect of the present invention is to provide a method and device that can rapidly estimate microbial contamination from different substrata, such as solid, soft or gelatinous surfaces, including fruits, vegetables, general agricultural and mass consumption products.
  • substrata such as solid, soft or gelatinous surfaces, including fruits, vegetables, general agricultural and mass consumption products.
  • Another aspect of the present invention is to provide a method and device that rapidly detects and identifies both aerobic and facultative anaerobic microorganism.
  • Still yet another aspect of the present invention is a method and device that can detect any saprophytic and semi-saprophytic microorganisms from any hard or gelatinous substrate, whether it be natural or synthetic.
  • Yet another aspect of the present invention is a method and device that enable efficient removal and multiple analyses in a very short period of time.
  • a method of assaying microbial contamination of substrata includes the steps of providing a collection device for obtaining a sample of the microbial contamination, contacting the surface with the collection device, whereby the collection device simultaneously obtains a sample of at least a portion of aerobic and anaerobic microbial contamination of the substrata, and placing the collection device into a receptacle for facilitating bacterial growth.
  • the receptacle contains a quantity of growth media. The receptacle is incubated and a bacterial colony grown in the receptacle is assessed.
  • a device for assaying microbial contamination of substrata including a collection device for obtaining a sample of aerobic and anaerobic microbial contamination.
  • a receptacle facilitates bacterial growth and contains a quantity of growth media.
  • the collection device is shaped to fit into the receptacle such that the collection device has continuous contact with the media.
  • FIG. 1 is a top view of the collection device of the present invention.
  • Fig. 2 is a cross-section of the device taken along line I-I of Fig. 1.
  • FIG. 3 is a top view of the collection device and growth receptacle of the present invention.
  • Fig. 4 is a cross-section of the growth receptacle and device taken along line II-II of Fig. 3.
  • Fig. 5 is a graphical representation of the method of the present invention vs. prior art assay methods.
  • the present invention relates to a method and device for rapidly collecting, detecting and identifying both aerobic and facultative anaerobic microorganisms, and determining the intensity of microbial contamination that could constitute a source of disease for animals or humans.
  • the device of the present invention can detect any saprophytic and semi-saprophytic microorganism from any hard or gelatinous substrate, whether it be natural or synthetic. Examples of such surfaces are fruits, vegetables, meat (cooked or fresh), cheese, wood, plastic, metal, etc. it should be appreciated that collection of microorganisms from other surfaces is contemplated by the present invention.
  • the device 10 for assaying microbial contamination of substrata consists of a collection device 12 for obtaining a sample of microbial contamination.
  • Collection device 12 can be an adhesive tape having an adhesive surface 14 on a side that is designated to remove microorganisms from any substrata.
  • the collection tape 12 can be Scotch 375 ® manufactured by 3M of St. Paul, MN. It should be appreciated that although the collection device of the present invention is referred to as an adhesive tape, other types of adhesive collection devices are contemplated by the present invention.
  • collection tape 12 is sized and configured to fit within a receptacle 20.
  • receptacle 20 can contain a quantity of growth media 24.
  • the collection tape 12 is designed to have the same shape as the receptacle and is sized to fit within the receptacle such that the collection tape has continuous contact with media 24.
  • collection tape 12 includes a plurality of perforations or holes 16 located on one half thereof for growing aerobic bacteria therein.
  • the other half 18 of the collection tape is un-perforated for simultaneously growing the anaerobic bacteria.
  • the perforations are shown as being located on one-half of the collection tape, it should be appreciated that the perforations can cover a larger or smaller area on the tape. However, for favorable bacteria growth, the perforations should cover at least one half of the tape.
  • Perforations 16 can have a diameter in the range of 0.5 - 1.5 mm and can be located at a distance of approximately 5 mm from each other to allow for the maximum number of perforations on the one half of the tape.
  • receptacle 20 can be a standard Petri dish.
  • Collection tape 12 can be a disk of material sized and shaped to fit within the Petri dish.
  • Petri dishes having a diameter of 36 mm, 55 mm, 86 mm or more can be used.
  • General microbial analysis can be estimated using universal media rich in complex nutrient elements, for example, Sabouraud dextrose agar and yeast, Sabouraud dextrose agar supplemented with egg yolk and milk, blood agar, bacto-agar with beef extract, and bacto- agar with liver extract.
  • Other selective medias can be used to encourage the growth of specific groups or species of bacteria.
  • the method of the present invention allows for rapid assaying of microbial contamination of the substrata.
  • the collection tape 12 is tightly pressed on the surface to be tested.
  • the tape should be pressed firmly to insure that contact is made over the surface, especially when probing irregular surfaces.
  • the tape having simultaneously obtained a sample of at least a portion of aerobic and anaerobic microbial contamination of the substrata, is placed within the receptacle 20.
  • the receptacle facilitates bacterial growth by containing a quantity of growth media.
  • the amount of media contained within receptacle 20 depends upon the size of the receptacle. However, at least 1 ml of media or sterile water should be deposited on the surface of the collection tape to insure continuous contact and eliminate air bubbles.
  • the receptacle is then incubated.
  • Petri dish receptacles and tape should be incubated between 26-28 0 C. During incubation the bacterial colony is grown. Due to the construction of the collection device, aerobic and anaerobic contaminants will simultaneously grow under the perforated and un-perforated areas respectfully.
  • the method of the present invention can estimate the microbial contamination of solid, soft or gelatinous substrata. More particularly, microbial contamination of fruits, vegetables, agricultural and/or mass consumption products.
  • the number of bacterial colonies determined can be assessed to indicate the degree of contamination.
  • the degree of contamination can be scaled. For example, less than 90-100% of perforation having microbial colonies could correspond to a contamination level 10; 90-80% - level 9; 80-70 - level 8; 70-60% - level 7; 60-50% - level 6; 50-40% - level 5; 40-30% - level 4; 30-20% - level 3; 20-10% - level 2; and 10-1% contamination - level 1.
  • Further identification of the microorganisms can be accomplished using several different methods. For example, traditional, i.e., morphological, serological, immunofluorescent, and morphological analyses, and modern, i.e., DNA analysis.
  • Fig. 5 is a graphical representation of the average number of bacteria that can be removed from substrata using the device of the present invention as compared to the prior art imprint and wash assay methods and devices. As can be seen from the graph, the rapid assay method and tape of the present invention had a ten-fold higher detection capacity than the prior art methods.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Clinical Laboratory Science (AREA)
  • Analytical Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé et un dispositif permettant une estimation rapide de la contamination microbienne de substrats différents, tels que des surfaces solides, molles ou gélatineuses, y compris des fruits, des légumes et des produits de grande consommation du secteur agricole. Le dispositif comprend : un dispositif collecteur pour l'obtention d'un échantillon d'une contamination microbienne aérobie et non aérobie; un réceptacle destiné à faciliter la croissance bactérienne, chaque dispositif collecteur étant d'une forme permettant son ajustement dans ledit réceptacle.
PCT/US2008/051631 2007-01-22 2008-01-22 Procédé et dispositif d'analyse rapide permettant d'estimer la contamination microbienne de substrats différents WO2008091851A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US88149307P 2007-01-22 2007-01-22
US60/881,493 2007-01-22

Publications (1)

Publication Number Publication Date
WO2008091851A1 true WO2008091851A1 (fr) 2008-07-31

Family

ID=39409820

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2008/051631 WO2008091851A1 (fr) 2007-01-22 2008-01-22 Procédé et dispositif d'analyse rapide permettant d'estimer la contamination microbienne de substrats différents

Country Status (1)

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WO (1) WO2008091851A1 (fr)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5994149A (en) * 1997-10-01 1999-11-30 Leonard Bloom Rapid test employing an adhesive slide
US6022682A (en) * 1996-08-14 2000-02-08 Minnesota Mining And Manufacturing Company Article and method for detection of enterotoxigenic staphylococci
WO2002046354A2 (fr) * 2000-12-08 2002-06-13 3M Innovative Properties Company Mise en images et collecte automatisees de colonies microbiennes sur des dispositifs de culture de couches minces
US6602704B1 (en) * 2002-06-24 2003-08-05 Biomerieux, Inc. Sample contact plate with latchable cover
ES2204326A1 (es) * 2002-10-03 2004-04-16 Henkel Iberica S.A. Procedimiento para determinar la contaminacion microbiana de una superficie dura.

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6022682A (en) * 1996-08-14 2000-02-08 Minnesota Mining And Manufacturing Company Article and method for detection of enterotoxigenic staphylococci
US5994149A (en) * 1997-10-01 1999-11-30 Leonard Bloom Rapid test employing an adhesive slide
WO2002046354A2 (fr) * 2000-12-08 2002-06-13 3M Innovative Properties Company Mise en images et collecte automatisees de colonies microbiennes sur des dispositifs de culture de couches minces
US6602704B1 (en) * 2002-06-24 2003-08-05 Biomerieux, Inc. Sample contact plate with latchable cover
ES2204326A1 (es) * 2002-10-03 2004-04-16 Henkel Iberica S.A. Procedimiento para determinar la contaminacion microbiana de una superficie dura.

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