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WO2000066267A1 - Systeme empechant l'intercontamination dans une plaque multipuits - Google Patents

Systeme empechant l'intercontamination dans une plaque multipuits Download PDF

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Publication number
WO2000066267A1
WO2000066267A1 PCT/US2000/011505 US0011505W WO0066267A1 WO 2000066267 A1 WO2000066267 A1 WO 2000066267A1 US 0011505 W US0011505 W US 0011505W WO 0066267 A1 WO0066267 A1 WO 0066267A1
Authority
WO
WIPO (PCT)
Prior art keywords
plate
openings
matrix member
nozzles
wells
Prior art date
Application number
PCT/US2000/011505
Other languages
English (en)
Inventor
Darin P. O'brien
Ellen M. Heath
Original Assignee
Gentra Systems, 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 Gentra Systems, Inc. filed Critical Gentra Systems, Inc.
Priority to AU46767/00A priority Critical patent/AU4676700A/en
Publication of WO2000066267A1 publication Critical patent/WO2000066267A1/fr

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B01L3/50255—Multi-well filtration

Definitions

  • the present invention relates generally to sample processing equipment and more specifically to multi-well purification devices.
  • Multi-tier multi-well plate systems have a top plate in which the sample is placed to be purified and isolated.
  • the top plate has holes which permit the flow-through of the samples or waste from processing.
  • Multi-tier systems also have a collection reservoir, which is commonly either a vacuum manifold or a bottom plate with wells, to collect either the waste or the final product.
  • a collection reservoir which is commonly either a vacuum manifold or a bottom plate with wells, to collect either the waste or the final product.
  • Multi-well plates can be used to purify specific components from biological, environmental, or pharmaceutical samples. These components can be proteins, lipids, nucleic acid or carbohydrates as well as metabolytes or environmental elements, or combinations thereof.
  • One such use for multi-well plates is to purify deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from biological materials.
  • DNA is a nucleic acid molecule that is the carrier of coded genetic information.
  • DNA and RNA are used in many applications, including diagnosis of certain infections, forensic sciences, clinical applications, recombinant DNA research, cloning, sequencing and the like. However, the DNA or RNA molecules need to be separated or purified from body tissue or fluid samples.
  • a problem that arises when using multi-well plates is that cross- contamination may occur among adjacent wells of the plate.
  • the cross-contamination also occurs between the openings and nozzles of the flow-through top plate when the sample is exiting the plate.
  • Cross- contamination occurs when a sample from one well becomes mixed with a sample in another well. This compromises sample integrity and may lead to ruined samples and inaccurate and misleading diagnoses.
  • the cross- contamination can occur during many steps of the purification and testing process, such as when transferring samples between wells, adding a reagent, centrifuging, and the like.
  • the ability to process multiple samples at the same time without cross-contaminating the samples is crucial for clinical diagnostic and forensic laboratories. This is especially true for molecular diagnostics, where extremely sensitive PCR (Polymerase Chain Reaction) and RT-PCR assays and other amplification protocols employed can detect as little as a single molecule of cross-contamination.
  • U.S. Patent 4,680,269 to Naylor discloses a method to prevent cross- contamination in single plate systems in which the plate does not have a flow- through design.
  • Naylor describes a single plate antimicrobial test kit in which cross-contamination is prevented by use of an impregnated filter paper which covers the plate's well openings and is held down by a lid. The impregnated filter attracts and absorbs the volatile microbes.
  • the Naylor invention is only described in terms of a single tier multi-well sealable system, not multi-tier systems that have a top plate with a flow-through design, in which case the nozzles or openings are unsealable during processing.
  • Naylor requires an impregnated filter, which can increase the system's complexity. Therefore, the Naylor solution is not applicable to flow-through plates, because in such systems the nozzles cannot be sealed, and the lower plate's collection reservoir cannot be sealed, and that is a potential source of cross-contamination.
  • the present invention provides an easy to use system that prevents cross- contamination of samples in a multi-well testing system.
  • the system includes a multi-holed matrix member positioned against a flow-through plate of the system.
  • the plate has openings and nozzles that match and correspond to the openings in the matrix member.
  • the sample or waste product goes through the unsealed nozzles, while the matrix member prevents any cross contamination among wells or between nozzles.
  • a user can prevent any cross-contamination between the unsealed nozzles and wells of the flow-through plate.
  • the system includes a collection reservoir or base plate, to be used for DNA sample collection after it has been isolated in the top plate.
  • the matrix member also prevents cross-contamination among the base plate's unsealed wells during sample processing.
  • the present system provides the advantage of preventing cross- contamination of components such as nucleic acids in a multi-well plate kit used for purification and sample preparation.
  • the system is simple to use for multi-tier multi-well plate systems, advantageously providing sample and waste flow- through to an unsealed collection reservoir, while still preventing cross- contamination.
  • the system prevents cross-contamination during all the steps of the purification process, including centrifuging the multi-well plate, heating the plate, or vacuum aspirating the sample through the multi-well plate.
  • Fig. 1 is an exploded perspective view of one embodiment of a multi-well sample processing system according to the invention.
  • Fig. 2 is an assembled view of the system of Fig. 1.
  • Fig. 3 A is a top and side view of one embodiment of a flow-through plate according to the invention. The side view is along the line 3A-3A.
  • Fig. 3B is a top and side view of another embodiment of a flow-through plate according to the invention. The side view is along the line 3B-3B.
  • Fig. 4A is a top and side view of one embodiment of a matrix member according to the invention. The side view is along the line 4A-4A.
  • Fig. 4B is a view of another embodiment of a matrix member according to the invention applied to a plate.
  • Fig. 5 A is a top and side view of one embodiment of a base plate according to the invention. The side view is along the line 5A-5A.
  • Fig. 5B is a top and side view of another embodiment of a base plate according to the invention. The side view is along the line 5B-5B.
  • Fig. 5C is a top and side view of another embodiment of a base plate according to the invention. The side view is along the line 5C-5C.
  • FIGS 1 -5 show one embodiment of a multi-well sample processing system 100.
  • System 100 includes a flow-through top plate 105.
  • plate 105 contains ninety-six holes or openings 106.
  • At a bottom surface of plate 105 there is a plurality of drip directors or nozzles 107.
  • Each nozzle 107 is mounted directly beneath a corresponding hole or opening of flow- through plate 105.
  • Nozzles 107 control the flow-through rate and direction of waste product and samples through plate 105.
  • Figure 3B shows another embodiment of plate 105 without any nozzles.
  • the flow- through rate of samples can be regulated by the shape and size of holes 106 themselves.
  • System 100 also includes a filter or guard, such as matrix member 103 for absorbing volatile components, thus preventing cross-contamination among samples when they leave nozzles 107 or are in the wells of a collection reservoir, such as a vacuum manifold or a base plate 101.
  • a filter or guard such as matrix member 103 for absorbing volatile components, thus preventing cross-contamination among samples when they leave nozzles 107 or are in the wells of a collection reservoir, such as a vacuum manifold or a base plate 101.
  • Figure 4A shows one embodiment of a matrix member 103.
  • matrix member 103 is made from a filter material such as Whatman Chromatography Paper 3mm Chr (Cat.
  • Matrix member 103 is cut into a shape substantially equivalent to the shape and dimensions of plate 105, in this embodiment, for example, the shape would be approximately 8 cm x 12 cm.
  • Matrix member 103 has a plurality of openings 104 which match and correspond to openings 106 in plate 105.
  • Openings 104 are substantially the diameter of the outside diameter of nozzles 107, approximately 1/8 inch in diameter, although those skilled in the art will recognize that other sizes can be used depending on the size of the nozzles or plate openings. Openings 104 are punched to correspond to the 8 x 12 ninety-six well format described above for the top plate. In this embodiment, openings 104 in matrix member 103 surround drip directors 107. When matrix member 103 is positioned against plate 105 the matrix member 103 intercepts and absorbs volatile nucleic acids leaving nozzles 107, or coming up from the wells of plate 101.
  • openings 104 in matrix member 103 are cut to a size slightly smaller than the diameter of nozzles 107. This permits matrix member 103 to be positioned against plate 105 by pushing the nozzles through the openings of the matrix member. Because the openings are slightly smaller than the nozzles, the matrix member is seated in position such that the matrix member surrounds the base of nozzles 107.
  • matrix member 103 can have an adhesive, such as 3M Super 77 Spray Adhesive (manufactured by 3M, St. Paul, MN), applied to the side which is to face the top plate. When pressed against the plate, the adhesive holds the matrix member in position.
  • Figure 4B shows another embodiment of a matrix member 403.
  • the matrix member includes a series of horizontal strips 401 and a series of vertical strips 402.
  • Strips 401 and 402 are made from the same material as the matrix member described above. However, in this embodiment, the material is cut in strips and then adhesively applied to plate 105. Strips 401 and 402 are applied so that each nozzle 107 is surrounded by absorbent material.
  • FIG. 5 A shows a top and side view of one embodiment of a receptacle plate or base plate 101.
  • Base plate 101 has a plurality of wells 102.
  • Wells 102 provide a cavity or space to capture waste from the samples or a place to capture a purified sample from nozzles 107.
  • Wells 102 run down substantially the whole depth of plate 101 as shown in Figure 5 A.
  • base plate 101 has ninety-six wells in an 8 x 12 configuration, corresponding and matching the configuration of matrix member openings 104 and top plate openings 106.
  • Each well 102 has a well opening 108 at the top surface of base plate 101.
  • FIG. 5A shows a base plate.
  • FIG. 5B shows a base plate.
  • a base plate well 508 includes an individual tube 509, thus permitting the user to individually handle collected samples.
  • no base plate is necessarily needed as a waste collection reservoir because the top flow-through plate and matrix member assembly can also be used above a vacuum manifold or other conduit to collect waste product.
  • matrix member 103 is positioned and mounted against the surface of flow-through top plate 105 containing nozzles 107. As described above, the matrix member may be either press fit or adhesively mounted against the plate.
  • the plate/matrix member assembly is next positioned upon a first base plate 101, as shown in Figure 2. Samples from which DNA is to be isolated, such as from whole blood, bone marrow, buffy coat, or body fluids, are placed in each opening 106 of plate 105. A purification solution is added to each opening. A seal is placed over the top openings of the top plate. The completed two-plate assembly is then centrifuged. More purification solution is added and the assembly is centrifuged again.
  • the system and process described above was effective in preventing cross-contamination during a test of the system. It was tested by application of the matrix member to a GENERATION ® Capture Plate (Gentra Systems, Inc., Minneapolis, MN) used for the batch purification of genomic DNA from whole blood from 96 samples simultaneously in a 96-well plate. Two plate systems were ran simultaneously, one with the matrix member applied and one without the matrix member. To test for cross-contamination, 200 ⁇ l of whole blood were loaded in alternate wells of the top flow-through plates starting with the first well in the first row of the plate. Wells not loaded with blood were loaded with a solution not containing DNA, phosphate buffered saline.
  • the system described above provides the advantage of preventing cross- contamination in a multi-well kit.
  • the system is simple to use in multi-tier multi-well plates, providing flow-through access to a collection reservoir while still preventing cross-contamination between unsealed nozzles and wells of the flow-through top plate and between wells of the unsealed collection reservoir.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un système empêchant l'intercontamination dans un système de traitement d'échantillon multipuits. Le système selon l'invention comprend un élément de matrice absorbant ou de filtre à trous multiples, placé entre une plaque à circulation continue supérieure et une plaque collectrice inférieure du système. La plaque supérieure comporte des ouvertures et des buses et la plaque inférieure comporte des ouvertures correspondant aux ouvertures dans l'élément de matrice. L'échantillon peut traverser les ouvertures et buses de la plaque supérieure en direction des puits de la plaque inférieure, mais l'élément de matrice empêche l'intercontamination entre les puits de la plaque inférieure ou entre les buses. Ainsi, un utilisateur peut empêcher l'intercontamination entre les buses non étanches de la plaque supérieure et les puits non étanches de la plaque inférieure.
PCT/US2000/011505 1999-04-30 2000-04-28 Systeme empechant l'intercontamination dans une plaque multipuits WO2000066267A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU46767/00A AU4676700A (en) 1999-04-30 2000-04-28 Preventing cross-contamination in a multi-well plate

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US30285799A 1999-04-30 1999-04-30
US09/302,857 1999-04-30

Publications (1)

Publication Number Publication Date
WO2000066267A1 true WO2000066267A1 (fr) 2000-11-09

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Application Number Title Priority Date Filing Date
PCT/US2000/011505 WO2000066267A1 (fr) 1999-04-30 2000-04-28 Systeme empechant l'intercontamination dans une plaque multipuits

Country Status (2)

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AU (1) AU4676700A (fr)
WO (1) WO2000066267A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003028878A1 (fr) * 2001-09-28 2003-04-10 Dynametrix Limited Procedes et moyens permettant de creer des arrangements
DE112006000361B4 (de) * 2005-02-18 2012-06-06 National University Corporation Saitama University Verfahren zur Einführung und Überführung einer Vielzahl kleinster Probenmengen

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4895706A (en) * 1986-10-28 1990-01-23 Costar Corporation Multi-well filter strip and composite assemblies
DE29722473U1 (de) * 1997-12-19 1998-02-19 Macherey, Nagel GmbH & Co. Handelsgesellschaft, 52355 Düren Trenneinrichtung zur Trennung von Substanzen
US5741463A (en) * 1993-04-19 1998-04-21 Sanadi; Ashok Ramesh Apparatus for preventing cross-contamination of multi-well test plates
DE19652327A1 (de) * 1996-12-16 1998-06-18 Europ Lab Molekularbiolog Vorrichtung zur Durchführung chemischer Reaktionsfolgen
WO2000025922A2 (fr) * 1998-10-29 2000-05-11 The Perkin-Elmer Corporation Appareil de microfiltration a plusieurs puits

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4895706A (en) * 1986-10-28 1990-01-23 Costar Corporation Multi-well filter strip and composite assemblies
US5741463A (en) * 1993-04-19 1998-04-21 Sanadi; Ashok Ramesh Apparatus for preventing cross-contamination of multi-well test plates
DE19652327A1 (de) * 1996-12-16 1998-06-18 Europ Lab Molekularbiolog Vorrichtung zur Durchführung chemischer Reaktionsfolgen
DE29722473U1 (de) * 1997-12-19 1998-02-19 Macherey, Nagel GmbH & Co. Handelsgesellschaft, 52355 Düren Trenneinrichtung zur Trennung von Substanzen
WO2000025922A2 (fr) * 1998-10-29 2000-05-11 The Perkin-Elmer Corporation Appareil de microfiltration a plusieurs puits

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003028878A1 (fr) * 2001-09-28 2003-04-10 Dynametrix Limited Procedes et moyens permettant de creer des arrangements
DE112006000361B4 (de) * 2005-02-18 2012-06-06 National University Corporation Saitama University Verfahren zur Einführung und Überführung einer Vielzahl kleinster Probenmengen
US8664005B2 (en) 2005-02-18 2014-03-04 National University Corporation Saitama University Method for introducing and transferring multiple minute quantity samples

Also Published As

Publication number Publication date
AU4676700A (en) 2000-11-17

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