Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N1/00—Sampling; Preparing specimens for investigation
  • G01N1/02—Devices for withdrawing samples
  • G01N1/22—Devices for withdrawing samples in the gaseous state
  • G01N1/2202—Devices for withdrawing samples in the gaseous state involving separation of sample components during sampling
  • G01N1/2205—Devices for withdrawing samples in the gaseous state involving separation of sample components during sampling with filters
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
  • C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
  • G01N33/56961—Plant cells or fungi
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N1/00—Sampling; Preparing specimens for investigation
  • G01N1/02—Devices for withdrawing samples
  • G01N1/22—Devices for withdrawing samples in the gaseous state
  • G01N1/2273—Atmospheric sampling
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N1/00—Sampling; Preparing specimens for investigation
  • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
  • G01N1/40—Concentrating samples
  • G01N1/405—Concentrating samples by adsorption or absorption
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N1/00—Sampling; Preparing specimens for investigation
  • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
  • G01N1/40—Concentrating samples
  • G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
  • G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
  • G01N2333/37—Assays involving biological materials from specific organisms or of a specific nature from fungi

Definitions

• This invention relates to an apparatus for and method of detecting an airborne mycotoxin in an enclosure and, more especially, an apparatus for and method of detecting an airborne trichothecene in a building.
• Mycotoxins are chemical substances produced by fungi.
• trichothecenes are known to be produced by a number of different fungi, such as Stachybotxys sp. and Fusarl ⁇ m sp.
• Stachybotxys sp. and Fusarl ⁇ m sp.
• Anguidine a type A trichothecene preparation called "Anguidine” was injected into humans.
• the subjects developed central nervous system and dermal disorders as well as other disorders. These symptoms are similar to those reported by occupants of mold contaminated buildings. It must be noted, however, that many factors can contribute to these symptoms.
• SBS Sick Building Syndrome
• Fungi and their secondary metabolites; such as, mycotoxins have been areas that have been closely examined. Fungi and their spores are known human allergens, especially in sensitized individuals. Fungi isolated from sick buildings include a wide variety of genera and species. It is logical to hypothesize that fungi growing indoors have been introduced from the outside. The fungi eventually find an effective growth niche consisting of water and an appropriate food source. The end result is an undesirable high concentration of fungal effluents. Of particular importance is that highly respirable fungal fragments are aerosolized simultaneously with spores in amounts as high as 320 times that of spores . These fungal fragments could be potential carriers of mycotoxins.
• SC Stachybotrys chartarvm
• an apparatus for detecting the presence of an airborne mycotoxin in an enclosure Pumping apparatus draws a portion of environmental air from the enclosure. A medium is disposed to receive the portion of the environmental air and to separate mycotoxins from the portion of air. Testing apparatus is then used to determine the presence of mycotoxins separated from the portion of air.
• a method of detecting the presence of an airborne mycotoxin in an enclosure comprises the steps of continually removing a portion of environmental air from a chosen area in the enclosure. The portion of environmental air is then passed through a filtering medium. Any filtrate filtered from the air is tested to determine the presence of a mycotoxin.
• FIG. 1 illustrates an apparatus for and a method of detecting the presence of an airborne mycotoxin in an enclosure or a building.
• FIG. 1 there is shown an apparatus 10 for and a method of detecting the presence of an airborne mycotoxin in an enclosure or building.
• a pumping apparatus 12 is used to draw a portion of the environmental air, as represented by arrow 14, from a location in the building or other enclosure and a filter medium 16 is disposed to receive the portion of the environmental air 14 and to separate and trap the airborne mycotoxins from the portion of air 14.
• the filtered air portion, as represented by arrow 18, may then be returned to the building or exhausted outside of the building.
• pumping apparatus 12 and filter medium 16 are shown as separate apparatus, they may be combined in a single housing and disposed within a single room of a residential house.
• the filter medium 16 may be a conventional electrostatic filter or may be made of particulate matter having a liquid provided on the surface of the particulate matters to absorb the mycotoxin while the air is flowing through the filter.
• a unitized machine that has been found to provide acceptable results is the DeLonghi DAP 130 Air Purifier with electrostatic filters manufactured for this machine.
• this machine When this machine is used, it is normally operated at its "high” setting with the ionizer on and the filters collecting airborne particles. Normally, the "high" setting provides a flow rate of about 8000 liters per minute and has lower settings to provide lower flow rates. Thus, the machine has flow rates up to about 80Q0 liters per minute. To insure sufficient environmental air is passed through filter medium 16 to obtain a competent test, it has been determined that the collection time take up to 24 hours and this 24 hour period may extend up to 7 days.
• Filter medium 16 is submerged in lOOOmL of pyrogen free water in a sterilized glass beaker 20 capable of containing this amount of volume. Beaker 20 with submerged filter is then placed in distilled water and a sonic cleaning apparatus 22 is used separate the particulate matter from filter medium 16.
• a sonic cleaning apparatus 22 is used separate the particulate matter from filter medium 16.
• beaker 20 is removed from ultrasonic cleaner 22 and allowed to sit at a room temperature of about 25°C between 18 and 24 hours.
• the filter medium 16 is removed from the water extract and squeezed to remove any absorbed water where it is collected in beaker 20.
• the filter extract is passed through sterilized Nalgene Reusable Filter Holders (Fisher Scientific Catalog number 09-740-23E) incorporating Whatman (Cat. No. 7402-004) 0.2 ⁇ m, 47mm nylon membrane filters.
• the cleanup filtrate 24 is accomplished using an in house vacuum operating at a flow rate of about 65 liters per minute.
• the cleanup filtrate is divided in two with each part being placed into VirTis 1200mL lyophilization jars. These samples were frozen using a rotating ethanol bath at -70°C on the Virtis Freezemobile. After the samples are frozen, they are lyophilized to dryness in the same machine, which incorporates a Fisher Scientific Maxima C Plus Model M6C vacuum pump. The two dried samples are suspended and combined in lOmL of a total pyrogen free water. The lOmL of concentrated filter extract 26 is again filtered. This time by being passed through Millex-GP 0.22pm Millipore sterilized syringe filters. The syringes used are Beaton Dickinson lOcc Luer SLIP TIP syringes.
• the final filtrate 26 is the working sample used in a conventional enzyme linked xmmunosorbent assay (" ⁇ LISA”) , such as that sold by "QuantiToxTM Trichothecenes Plate Kit. " This assay is manufactured by EnviroLogix of 500 Riverside Industrial Parkway, Portland ME 04103-1418. It is believed that this test kit uses the apparatus and monoclonal antibodies disclosed in U.S. Pat. No. 4,772,551.
• ⁇ LISA enzyme linked xmmunosorbent assay
• each well is rinsed five times with 300 ⁇ l of the phosphate buffered saline solution.
• the plate is then slapped on a paper towel to remove as much water as possible.
• 100ml of the substrate provided with the kit is added to each well.
• the contents of the wells are thoroughly mixed.
• the wells are covered with new tape or Parafilm and incubated for 15 minutes at ambient temperature.
• 100ml of a Stop Solution provided with the kit is added to the wells. The solution is then read at 450nm.
• ELISA inhibition rates ranged from 35.5% to 95.0% compared to controls.
• the presence of macrocyclic trichothecenes was confirmed using a modified Andersen Polyurethane Foam High Volume Air Sampler in one residence. Sampling times were 24, 48, and 72 hours.
• ELISA inhibition rates ranged from 70.0 to 79.1% with the first stage filters and increased significantly over time (27.1 to 49.4%) on the second stage filters.
• EXAMPLE I The first building selected was an unoccupied house that contained personal belongings. Two rooms were chosen for testing, the living room and the utility room. The living room had no visible fungal growth and was open to the rest of the house. The utility room was documented to have a leak from the water heater and was an enclosed area. Fungal growth was visible and was confirmed to be Stachybotrys chartarum (SC) .
• SC Stachybotrys chartarum
• the second building selected was an unoccupied house that contained personal belongings .
• An enclosed closet was chosen for testing. This was a storage closet in the garage. Fungal contamination was visible. SC was confirmed among other organisms.
• a DeLonghi DAP 130 Air Purifier with an electrostatic filter in place was set at high was positioned at floor level in the storage closet. The purifier was operated at environmental temperatures and pressures. Air conditioning was turned off and never on in the houses during testing. Even though the purifier came equipped with pre-filters for large particles, the pre- filters were removed before testing began. The purifier ran for one week. The electostatic filters were removed from the machine and handled in accordance with the procedure previously described. After the final filtrates were obtained, they were processed in accordance with the procedure relating to the ALISA previously described to indicate the presence of a trichothecene (a mycotoxin) within the building.
• EXAMPLE III The third building selected was a house that was occupied, but the room chosen for testing was enclosed and remained closed to the rest of the house. The room was a bathroom. Fungal contamination was visible in the shower. SC was confirmed among other organisms.
• a DeLonghi DAP 130 Air Purifier with an electrostatic filter in place was set at high and positioned at about two feel above floor level in the bathroom. The purifier was operated at environmental temperatures and pressures . Air conditioning was not turned off. Even though the purifier came equipped with pre-filters for large particles, the pre-filters were removed before testing began. The purifier ran for one week. The electostatic filters were removed from the machine and handled in accordance with the procedure previously described.
• EXAMPLE IV The fourth building selected was an unoccupied house that contained personal belongings .
• Four rooms were chosen for sampling - the living room, TV room, upstairs bedroom, and kitchen. No room was entirely closed off to the rest of the house.
• the living room was sampled for 24 hours with the purifier being at floor level.
• the other three rooms were sampled for one week with the purifier in the TV room being elevated above floor level by about 3.5 feet, the purifier in the upstairs bedroom being elevated above the floor level by about two feet and the purifier in the kitchen being at floor level.
• Fungal contamination was clearly evident in all pf the rooms.
• the kitchen showed the heaviest fungal growth. SC was confirmed among other organisms.
• a DeLonghi DAP 130 Air Purifier with an electrostatic filter in place was operated at environmental temperatures and pressures.
• the fifth building selected was an unoccupied house that contained no personal belongings.
• Four rooms were chosen for sampling - the main entry room, the back entry room, the kitchen, and a bedroom.
• the house was open to the outside environment (much of the roof was not present, only covered by a tarp, and some of the floor had been removed and exposed to the foundation) so environmental conditions most likely varied. These, however, were not measured.
• the chosen bedroom was sampled for 24 hours at floor level.
• the main and back entry rooms were sampled for one week at floor level.
• the kitchen was sampled for one week at an elevation of about 4 feet above floor level. No room was closed off to the rest of the house. Fungal contamination was clearly evident throughout the house, the worst being the kitchen. SC was confirmed.
• a DeLonghi DAP 130 Air Purifier with an electrostatic filter in place was operated at environmental temperatures and pressures at each of the speci ied locations for the specified periods of time. Even though the purifier came equipped with pre- filters for large particles, the pre-filters were removed before testing began. The purifier in the corner room ran for 24 hours and the remaining purifiers ran for one week. The electrostatic filters were removed from the machine and handled in accordance with the procedure previously described. After the final filtrates were obtained, they were processed in accordance with the procedure relating to the ALISA previously described to indicate the presence of trichothecene (a mycotoxin) within the building.

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• 2003
  • 2003-05-12 WO PCT/US2003/014856 patent/WO2004106933A1/en not_active Ceased
  • 2003-05-12 EP EP03741800A patent/EP1625401A4/de not_active Withdrawn
  • 2003-05-12 AU AU2003304160A patent/AU2003304160A1/en not_active Abandoned

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