WO1990013624A1 - Systeme de culture de croissance de microorganismes, procede et unite de filtrage utilises dans un tel systeme - Google Patents

Systeme de culture de croissance de microorganismes, procede et unite de filtrage utilises dans un tel systeme Download PDF

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Publication number
WO1990013624A1
WO1990013624A1 PCT/US1989/001965 US8901965W WO9013624A1 WO 1990013624 A1 WO1990013624 A1 WO 1990013624A1 US 8901965 W US8901965 W US 8901965W WO 9013624 A1 WO9013624 A1 WO 9013624A1
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WO
WIPO (PCT)
Prior art keywords
section
vessel
fluid
filtration unit
filter membrane
Prior art date
Application number
PCT/US1989/001965
Other languages
English (en)
Inventor
Michael P. Friedman
James Jablonsky
Ronald F. Schell
Original Assignee
Human Medical Laboratories, Inc.
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 Human Medical Laboratories, Inc. filed Critical Human Medical Laboratories, Inc.
Priority to PCT/US1989/001965 priority Critical patent/WO1990013624A1/fr
Publication of WO1990013624A1 publication Critical patent/WO1990013624A1/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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/02—Membranes; Filters
    • 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
    • C12M23/32—Frangible parts
    • 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
    • C12M23/34—Internal compartments or partitions
    • 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
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
    • 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

Definitions

  • the present invention is concerned with an improved apparatus and method for detecting, identifying and testing microorganisms isolated from liquids, particularly but not necessarily blood or other body fluids.
  • Bacteremia is a clinically significant, potentially life-threatening event.
  • the rate of mortality associated with bacteremia remains high.
  • Recovery from bacteremia is increased when appropriate antimicrobial therapy is instituted early in the course of infec ⁇ tion. Therefore, prompt detection, identification and suscepti ⁇ bility testing of microorganisms isolated from blood is imperative.
  • Procedures for diagnosing bacteremia require rapid isolation of the infecting organism. Culturing blood in a growth medium using commercially available blood culture bottles is the standard isolation technique. However, this technique has several disadvantages. Thus, for example, detection can be affected by the type of medium, the density of bacteria in blood, the time of collection, the volume of blood, the host's immune response,, laboratory practices and other factors. In addition, several hours or days may be required to obtain results, depend ⁇ ing on the bacteria's growth characteristics.
  • the present invention offers an improved method and apparatus for detecting and identifying microorganisms in blood or other body fluids or the like which avoid or substantially reduce the number of problems encountered with previously available approaches.
  • a number of the advantages of the present method and apparatus are detailed hereinafter. Broadly speaking, however, the invention provides a relatively quick, simple and accurate bacterial culture system for isolating and identifying microorganisms, particularly those isolated from the blood or other body fluids.
  • the invention is based, to a significant extent, on a special combination and adaptation of centrifugation, filtration and growth media techniques which cooperate to expedite the recovery and identification of microorganisms from blood or the equivalent.
  • fresh whole human blood is mixed with a fluid density gradient in a specially designed collection tube generally similar to the conventional vacutainer.
  • the tube is centrifuged in standard manner for a suitable period of time, e.g , 30 minutes or so, to separate erythrocytes by sedimentation.
  • the remaining fluid plasma, leukocytes and fluid gradient
  • a media cup containing a selected growth of culture medium is attached to the base of the filter to promote bacterial growth.
  • the collected microorganisms are then detected on the filter within an appropriate culture period, e.g., 18-24 hours.
  • the collection tube and filter unit are so designed that the tube may be sealed into the filter unit for application of the filtration vacuum before the tube contents are open to the vacuum whereby possible contamination of the fluid is kept to a minimum.
  • the media cup likewise has a special sealing connection to the filter unit and the filter membrane is tightly sealed within the filter unit itself to avoid contamination. Additionally, the media cup is positioned close to, i.e., just below and functionally in contact with, the filter membrane so that the microorganisms collected on the filter membrane can be cultivated essentially "in situ".
  • the system of the invention is effective and sensitive for the recovery and detection of organisms in blood fluids or the like even when the organisms are present in very small amounts.
  • Tests conducted with seeded or inoculated human blood, using concentrations of inocular in the range of from 3.0 ⁇ 0.7 to 18.0 ⁇ 3.6 organisms per ml have been successfully conducted and indicate no statistically significant differences detectable between the number of microorganisms recovered by the present system and by the conventional culture of the original inoculum.
  • the invention is useful in detecting any type of known microorganism larger than viruses which may be found in the analysis of blood, other body fluid or the like.
  • Typical of such microorganisms are Candida albicans, Escherickia coli, Haemophilus influenzae, Klebsiella pneumoniae, Listeria monocytogenes, Neisseria meningitides, Pseudomonas aeruginosa, Staphylococcus aurias, Streptococcus faecalis, and Streptococcus pneumoniae. These microorganisms have been detected on the filter membrane, using the method and apparatus of the invention, within 18 hours after filtration. Detection times will vary depending on various factors, for example, the amount of microorganisms present.
  • SUBSTITUTESHEET a minimum of such manipulations and offers major advantages over previous techniques involving centrifugation and filtration. For example, no lysing agents, multiple filters, dilutions, sophisticated equipment or inordinate centrifugation speeds are required or involved. Clogging of filters because of contamination with leukocytes and erythrocytes is eliminated and a minimum amount of time, e.g., less than 30 seconds, is required to filter the entire gradient.
  • the blood collecting tube, filter and media are designed to prevent contamination, the overall system being effectively sealed.
  • Fig. 1 is a perspective view of the collecting tube
  • Fig. 2 is a top plan view of the apparatus
  • Fig. 3 is a vertical sectional view of the apparatus on the line 3-3 of Fig. 2;
  • Fig. 3A is a perspective view of a cup member for use with the apparatus
  • Fig. 4 is a side view of the filter and media units of the apparatus showing how these units stack together;
  • Fig. 5 is a vertical sectional view through the filter and media units
  • Fig. 6 is a fragmented view, partly in section, showing individual parts of the filter unit
  • Fig. 7 is an enlarged partial section of the filter unit showing how the several parts fit together.
  • Fig. 8 is a perspective view showing separate parts of the filter unit.
  • the culture system of the invention comprises a collecting and separating device (C) ; a filtration unit (F) ; and media cup (M) .
  • the collecting and separating device (C) is similar to the known "vacutainer" sampling system. It comprises a cylindri ⁇ cal glass tube (2) which is preferably composed of boro-silicate glass; a stopper (4) of butyl rubber or the like closing the top inlet end of the tube; and a vacuum coupling separation device, broadly referred to by the numeral (5) , closing the other end.
  • the device (5) may comprise a rubber stopper (6) or the equivalent provided with a central passageway (7) which is narrowed at its inner end to support a hollow open-ended needle (8) whose inner end projects into tube (2) and is apertured as shown at (10) .
  • the other end of needle (8) empties into an enlarged zone (12) of passageway (7) .
  • a breakable membrane (14) normally extends across the passageway (7) to seal the needle (8) and interior (15) of tube (2) and avoid contamination of the tube contents.
  • a removable seal tab (16) is also placed over the end of the tube to close passageway (7) .
  • Tube (2) is filled with a fluid gradient or solution that can be used to collect the fluid containing the organisms to be studied.
  • the fluid gradient in tube (2) is such that, for example, on centrifuging blood, erythrocytes (red blood cells) are separated by sedimentation and the remaining fluid can be used for test purposes.
  • the nature and composition of the gradient fluid in tube (2) may be widely varied and will be suitably selected depending on the fluid to be analyzed.
  • the tube, with fluid gradient therein is kept in the sealed, pre-sterilized condition as shown in Fig. 1 with tab (16) thereon until it is to be used.
  • Blood or like fluid to be analyzed may be added to the tube by means of a sterile needle or the like via the top end of the tube through stopper (4) or after its removal.
  • Components B, C and D are then mixed together after which 800 ml of this mixture are blended with 300 ml of the warm 85% Hypague (Component A) .
  • the density of the solution is advan ⁇ tageously kept around 1.149.
  • whole human blood is mixed with 10 ml of the above formulation in tube (2) , the blood being added through stopper (4) .
  • the resulting mixture is then centrifuged at 2000 RPM (Sorvall Instrument GLC-4, a general laboratory centrifuge with rotor 4-1000) for 30 minutes.
  • centrifuging can be carried out in conventional manner using standard non-refrigerated table top centrifuges. Centrifuging speeds in the order of 2000 to 10,000 RPM for 15 to 40 minutes usually are effective to provide the desired separation of erythrocytes although it will be appreciated that the centrifuging speed and time involved can be varied depending, for example, on the fluid being centrifuged for analysis.
  • the tube (2) is sealed into the assembled filtration unit (F) to filter out the organisms from the centrifuged fluid as shown in Fig. 3. More particular ⁇ ly, the tab (16) is removed from the tube after centrifugation is completed and the tube is then set on the top face of the filter unit (F) as shown in Fig. 3. To this end, the top surface (17) of the filter unit is provided with a circular pocket or indent (18) which is adapted to receive the tube (2) as shown. This pocket (18) is off-center with respect to the top surface of the filter unit as shown in Fig. 2. A purpose of this is to
  • SUBSTITUTE SHEET facilitate inspection of the interior of the filter unit, particularly the filter membrane, even when the tube is inserted into the unit as shown in Fig. 3.
  • a feature of the invention is that the filter unit, and preferably also the media cup, are made from transparent molded plastic so that it is possible to easily observe the filtering of the microorganisms and their growth on the filter as herein described.
  • the pocket or indent (18) is provided with a centrally located hollow projection (20) which can be used to break the membrane (14) when the tube (2) , after centrifugation, is placed on the filter unit.
  • the upper end of the hollow projection which may be provided with a razor-like cutting edge, pierces the membrane and opens up communication between the contents of tube (2) and the filter unit (F) . Because of the snug fit which is provided between the tube and the indent or pocket (18) , the tube contents can be effectively sealed off from contamination by the surroundings while remaining open to communication with the filter unit to effect the desired filtering operation.
  • Fig. 3 the tube (2) is shown incompletely inserted into the indent or pocket (18) in order that the membrane (14) might also be illustrated. However, it will be appreciated that when the tube (2) is pushed all the way into the pocket (18) , the membrane (14) is broken by projection (20) to open up the necessary communication between the interior (15) of tube (2) and the filter unit (F) .
  • the function of filter unit (F) is to permit separation of the microorganisms by vacuum in a sealed noncontam- inating environment and to support the separated organisms on the filter so that they may grow in a sealed, noncontaminable environment.
  • the unit (F) comprises three parts, namely, the top section (21) ; the middle section (22) which, with section (21) holds the filter (23) in place; and the bottom section (24) which connects with the vacuum source, broadly shown at (25) .
  • the top section (21) is the unit member which is provided with the pocket (18) to receive the tube (2) in a sealed
  • a removable cup member (26) or the like may be provided to cover the projection (20) when the filter unit (F) is not joined to the tube (2) or otherwise not in use.
  • the bottom surface of the top section (21) is provided with a pair of spaced downwardly extending ribs (27) and (28) which extend peripherally around the bottom of member (21) and are adapted to mate with an appropriately shaped upwardly extending rib (29) which extends around the upper periphery of middle section (22) .
  • This arrangement permits the filter (23) to be held firmly in place by the mating of the members (21) and (22) as shown in Figs. 3 and 5.
  • the peripheries of the opposed surfaces of the middle section (22) and the bottom section (24) are also mated as shown in Fig. 3 so as to provide a compact, rigid unit (F) when the three sections are placed together.
  • top section (21) of the filter unit (F) is essentially square in shape although other shapes, e.g., circular or oval, may be used.
  • the middle and bottom sections may have the same shape as the top section or they may have different shapes provided the three sections can be mated together to provide a compact unit with the filter (23) in an essentially sealed condition.
  • the filter (23) preferably comprises a support screen (30) having a circumferential trough ⁇ like periphery (32) which is adapted to receive a top grid or screen (34) , the edge of the screen being shaped to fit into the support screen with a paper filter (36) firmly held therebetween as best shown in Fig. 7.
  • both the support screen (30) and the grid screen (34) are made from rigid plastic of sufficient strength and rigidity to hold the paper filter (36) in place and withstand the vacuum imposed during filtration and possible backflushing should the latter be desired.
  • the mesh sizes of the grid screen and support screen are not critical. The two screens obviously should be sufficiently open to permit the filtration to take place
  • the bottom section (24) of the filtration unit (F) is provided with means (25) through which a vacuum may be drawn when the tube (2) and filtration unit have been assembled.
  • a vacuum may be drawn when the tube (2) and filtration unit have been assembled.
  • Preferable means for example, a removable tear or seal tab (not shown) , is used to cover the vacuum connection to prevent contamination when the unit (F) is not in use, the tab being removed when the vacuum connection is to be made.
  • filtration units (F) It is convenient to store the filtration units (F) in sterile condition with the three sections (21) , (22) and (24) and the paper filter (23) assembled ready for use > and with the top and bottom sealed by removable tabs, as indicated.
  • a number of such units may also be stacked together generally in the manner shown in Fig. 4 although this particular Fig. is primarily intended to illustrate how the units (F) , with media cups (M) attached, can also be stacked to cultivate the media.
  • the flat outer periphery of each filter unit lends itself to marking so that, for example, the source of a fluid being analyzed or other relevant information can be placed on the surface.
  • the filtration unit, and preferably the media cup as well are made from transparent plastic, it is possible to easily see what is going on in any particular unit or stack of units.
  • the removable cup member or seal (26) is removed and a tube (2) , after centrifugation and removal of the seal (16) therefrom is inserted into the pocket (18) of the filter unit as shown in Fig. 3, the tube being fully pushed down into the pocket to cause the projection (20) to break the membrane (14) .
  • the bottom seal (not shown) of the filter unit is then removed and the unit is attached to a suitable vacuum source. Vacuum is then drawn on the system and the supernatant liquid in the tube, freed of erythrocytes in the case of blood, is drawn from the tube (2) through the hollow needle (8) onto and through the filter paper (23) .
  • the bottom section (24) of the filtration unit which acts as a general purpose receptacle for effluents passing through the filter, is replaced by a growth media cup or dish (M) .
  • Cup (M) may have generally the same outer configuration as the bottom section (24) of the filtration unit so that it mates snugly with middle section (22) as shown in Fig. 5.
  • the elements (24 and M) are preferably cylindrical to differentiate from Sections (21) and (22) of the filtration unit, which preferably have flat sides, to thus facilitate removal and insertion.
  • the upper peripheral surface thereof should mate with the middle section (22) of the filter unit in the same way as the removable bottom section (24) of the filter unit.
  • the bottom section (24) can be removed by pulling the same downwardly away from the middle section and the media cup (M) then inserted as shown in Fig. 5.
  • the growth media cup (M) contains an appropriate growth medium, of the type known in the art, so as to encourage growth of bacteria collected on the filter paper (23) . As shown in Fig. 5, the media cup is positioned very closely to the filter paper so that media in cup (M) will essentially contact the microorgan ⁇ isms and/or otherwise cause them to grow right on the filter paper.
  • the bottom section (24) of the filtration unit and the growth media cup (M) may be provided with serrations (38) or the like to facilitate removal.
  • the growth media cup should be kept in a pre-sterilized condition with the appropriate media therein for use.
  • Fig. 4 illustrates the stacking feature of the filtration units either with the bottom section (24) or with the media cup (M) .
  • the units may thus be stacked for storage or they may be stacked for cultivation of the microorganisms on the filter by means of the growth media.
  • each of the filtration units is provided with an upper protrusion (40) which is adapted to fit into a mating recess in the bottom of an adjacent unit, thereby facilitating stacking as shown in Fig. 4.
  • Typical media include the following:
  • agar content can usually ⁇ be varied from 1 to 15 g to accommodate for other conditions, it being essential for the filter to absorb nutrients relatively quickly.
  • the media cup (M) differs from the bottom section (24) of the filtration unit in that (M) is closed at the bottom, i.e., it does not have a fitting for the application of vacuum but instead is engineered to hold the media that acts by contact or mechanical infiltration and capillary action of gelatinous material and nutrients to the organisms trapped on the filter. In this way the organisms that have been filtered out are nourished through placement of the correct medium in its appropriate cup.
  • the only other substantive structural difference between media cup (M) and section (24) is that the bottom inner wall of section (24) is slanted inwardly towards the vacuum source (25) as shown at (42) . This facilitates the removal of fluid which passes through the filter and avoids the collection of residues in section (24) .
  • the media cup (M) is covered with a pull tab (not shown) to assure sterility of the nutrient material until it is to be used.
  • the components of the present system are desirably kept sterile and, as will be evident, the mating sections of the filter unit and the media cup (M) have integrated into them a double wall sealing system to insure minimal contamination in use.
  • the resulting totally closed system lends itself very readily to eliminating many of the extraneous organisms that are picked up on other systems.
  • the system When used to analyze blood, the system requires centrifugation to sediment the red cells before the ingredients of the tube are withdrawn by vacuum onto the filter surface of the filter unit.
  • the filter units are approximately 50 millimeters in width and are attached to a vacuum source that allows the clinical microbiologist to process
  • the bottom portion of the filtration device is removed and replaced with the specifically designed media cup as mentioned above.
  • the function of the media is to supply nutrients to the filter surface and allow the microorganisms that have been trapped during the filtration process to grow.
  • the media cup and filter which thus become one unit may then be placed in an incubator, stacked with other units, if desired, according to Fig. 4, at the appropriate temperature for the specific organisms, i.e., fungi, bacteria, Rickettsiae, large virus, etc. and examined as and when desired.
  • the above described system can be used to detect as little as one organism per ml of whole blood and'most microorgan ⁇ isms will be detected within a time period of 18-24 hours. While the analysis of blood is a particularly important area of use for the invention, a much wider area of use is contemplated.
  • the invention may be used to detect microorganisms as follows:
  • Organisms in all body fluid i.e., pleural, spinal, seminal, urine, etc.
  • Dairy products such as milk which contain organisms
  • E SHEET It has been noted when using the present system that growth initially occurs on the filter in forms of characteristic colonies. This saves a minimum of one step from conventional culture techniques and therefore an additional 12 to 24 hours up to 48 to 72 hours. Colonies on the filter may be used directly for rapid identification in any bacterial susceptibility test. Inhibitory substances may be washed away, thus allowing more effective evaluation of therapy. Differential or selective solid media or both may be used directly. Rough quantification of bacteria present is also possible on the filter. Direct recognition and possibly even identification of organisms on the filter without prior incubation may also be accomplished using monoclonal antibodies with certain indicator tags. Early recognition of multiple bacteremia is also possible.
  • the speed of detection of bacteremia with the present system (hereinafter called the "MOG" System) is particularly advantageous. Most microorganisms (Haemophilus influenzae, Streptococcus spp., etc.) can be detected on filters within 18 to 24 hours after filtration. This is in contrast to conventional techniques where bacteria are isolated from blood culture bottles 48 to 72 hours after inoculation.
  • the system is exceptionally sensitive. For example, small numbers of microorganisms (1 to 18 organisms/ml) can be recovered on the filters.
  • the sensitivity of the blood culture bottles is unclear. For example, thioglycolate or thiol broths are not suitable for the isolation of Pseudomonas and yeast from blood. Detection of bacteria in the blood culture bottle is affected by the type of medium.
  • Antimicrobial agents and other inhibitory substances are removed when blood is filtered and washed.
  • removal of anti ⁇ microbials requires addition of other ingredients to the blood culture bottle to inactive antimicrobials and contamination increases with each additional entry into the blood culture bottle.
  • Complement and specific and nonspecific bactericidal antibodies are also not removed or inactivated by the blood culture media.
  • the present system requires about 30 minutes for centrifugation, 15-20 seconds for filtration (removal of bacterial from blood) and 5 to 10 minutes to present the media cup to the filter. Bacteria are isolated within 24 hours. In contrast, after 24 hours of incubation, blood culture bottles are inspected visually (10 to 30 minutes) , subcultured (1 to 2 hours) , gram stained (1 to 2 hours) , examined daily 1 to 7 days) , resubcultured and restained.
  • the present system does not clog filters. Clogging of filters made past ceritrifugation-filtration systems cumber ⁇ some. Bacteria are easily isolated by the system.
  • the present system also will allow quantitation of bacteria in a patient's blood. This advantage is very important. The prognosis or development of resistance to therapy can be determined. The quantitation of bacteria also helps to determine whether contaminants are present. In contrast, the blood culture bottle can perform neither of these functions.
  • Filtered bacteria can be placed on different media. Certain organisms require more nutrients. In contrast, the blood culture bottle contains only one medium and it is not satisfactory for promoting growth of all bacteria. 10. With the present system, agents to remove all antimicrobials can be incorporated in the tube and media can be inoculated with material to determine within 2 to 3 hours if bacteria are resistant to antimicrobials. Such functions cannot be performed in a blood culture bottle since bacteria are not isolated when growing in the bottle.
  • the MOG system is rapid, simple and has negligible toxicity whereas bacteria cannot be recovered from all blood culture media using the bottle system. Additionally with the latter system, the media affects the growth of bacteria. It is estimated that there are over 200 separate blood culture media available commercially today.
  • Inoculate MOG System 1. Inoculate blood culture (5 min.) medium (5 min.)

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Abstract

Un appareil pour isoler et identifier des microorganismes compris dans du sang ou autre comprend un récipient collecteur tubulaire (C) fermé hermétiquement (4,5) aux deux extrémités mais comprenant à une extrémité des moyens (5) permettant de sortir du liquide dans ledit récipient par l'application d'un vide lorsque le joint placé à cette extrémité est fracturé. L'appareil comprend également une unité de filtrage à sections multiples (F) comprenant une première section (21) pourvue de moyens (18) pour recevoir ladite extrémité du récipient collecteur ainsi que des moyens disposés dans la première section pour la fracture du joint lorsqu'il est reçu par ladite section. Dans l'unité de filtrage une deuxième section (22) est emboîtée dans la première section (21), une membrane de filtrage (23) étant placée entre la première et la deuxième sections emboîtées. L'unité de filtrage est pourvue également d'une troisième section (24) emboîtée dans la deuxième section, ainsi que de moyens (25) permettant d'aspirer un vide à travers le fond de ladite troisième section (24).
PCT/US1989/001965 1989-05-08 1989-05-08 Systeme de culture de croissance de microorganismes, procede et unite de filtrage utilises dans un tel systeme WO1990013624A1 (fr)

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2681874A1 (fr) * 1991-10-01 1993-04-02 Biogir Materiel pret a l'emploi permettant la realisation d'etudes de toxicite sur des cellules en culture.
WO1994028110A1 (fr) * 1993-06-01 1994-12-08 Celsis International Plc Dispositif de detection
DE19801090A1 (de) * 1998-01-14 1999-07-15 Gerhard Haase Lysis-Filtrations-Assay LYFA
WO2002068580A3 (fr) * 2001-02-28 2003-12-24 Bio Merieux Inc Dispositif de filtration et de detection integre
FR2885140A1 (fr) * 2005-04-29 2006-11-03 Millipore Corp Procede de detection et de caracterisation de microorgarnismes sur une membrane.
WO2019048405A1 (fr) * 2017-09-06 2019-03-14 Merck Patent Gmbh Ensemble de filtration et procédé pour un test microbiologique
WO2019048402A1 (fr) * 2017-09-06 2019-03-14 Merck Patent Gmbh Ensemble de filtration et procédé pour un test microbiologique

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Cited By (16)

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FR2681874A1 (fr) * 1991-10-01 1993-04-02 Biogir Materiel pret a l'emploi permettant la realisation d'etudes de toxicite sur des cellules en culture.
EP0536022A1 (fr) * 1991-10-01 1993-04-07 Société Anonyme dite : BIOGIR Matériel prêt à l'emploi permettant la réalisation d'études de toxicologie et de pharmacologie sur des cellules en culture
WO1994028110A1 (fr) * 1993-06-01 1994-12-08 Celsis International Plc Dispositif de detection
US5811257A (en) * 1993-06-01 1998-09-22 Celsis International, P.L.C. Detection apparatus
DE19801090A1 (de) * 1998-01-14 1999-07-15 Gerhard Haase Lysis-Filtrations-Assay LYFA
WO2002068580A3 (fr) * 2001-02-28 2003-12-24 Bio Merieux Inc Dispositif de filtration et de detection integre
FR2885140A1 (fr) * 2005-04-29 2006-11-03 Millipore Corp Procede de detection et de caracterisation de microorgarnismes sur une membrane.
WO2006117676A3 (fr) * 2005-04-29 2007-03-15 Millipore Corp Procede de detection et de caracterisation de micro-organismes sur un filtre
JP2008538910A (ja) * 2005-04-29 2008-11-13 ミリポア・コーポレイション フィルター上の微生物の検出及びキャラクタリゼーション方法
WO2019048405A1 (fr) * 2017-09-06 2019-03-14 Merck Patent Gmbh Ensemble de filtration et procédé pour un test microbiologique
WO2019048402A1 (fr) * 2017-09-06 2019-03-14 Merck Patent Gmbh Ensemble de filtration et procédé pour un test microbiologique
CN111032198A (zh) * 2017-09-06 2020-04-17 默克专利股份公司 用于微生物测试的过滤组件和方法
CN111065446A (zh) * 2017-09-06 2020-04-24 默克专利股份公司 用于微生物测试的过滤组件和方法
CN111065446B (zh) * 2017-09-06 2022-09-06 默克专利股份公司 用于微生物测试的过滤组件和方法
US11654398B2 (en) 2017-09-06 2023-05-23 Merck Patent Gmbh Filtration assembly and method for microbiological testing
US11731083B2 (en) 2017-09-06 2023-08-22 Merck Patent Gmbh Filtration assembly and method for microbiological testing

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