JP2007515943A - Cage for sample collection and preparation - Google Patents

Abstract

  A holder for PCR sample collection and preparation, having a buffer container housing removably connected to a plunger housing, and a method for using the same. A swab attached to one end of the plunger takes a sample of the sample to be analyzed for the presence or absence of a biological warfare agent. The swab and the plunger are inserted into the plunger housing, the buffer container is disposed inside the buffer container housing, and the buffer container housing and the plunger housing are attached. The buffer passes through the swab to elute the sample and the sample is mixed with the reagents. The prepared sample is fed into the reaction tube by a whipping operation for analysis.

Description

(Cross-reference of related applications)
This application is a continuation-in-part of US patent application Ser. No. 10 / 727,037, filed Dec. 4, 2003, and is therefore US Provisional Application No. 60 / 430,994, filed Dec. 4, 2002. You have the right to benefit from the issue. This application also claims the priority and benefit of US Provisional Application No. 60 / 473,539, filed May 28, 2003. All of the above three applications are incorporated herein by reference in their entirety.

  This application relates to a system designed to detect polymerase chain reaction (PCR). The system includes a device designed to detect the presence or absence of a biological agent through DNA amplification. Detection is accomplished through heating and cooling of the PCR reaction mixture placed in a disposable reaction tube. The presence of the biological agent to be detected is indicated by an increase in fluorescence emission.

  The present invention relates to a holder for polymerase chain reaction (PCR) collection and preparation comprising a plunger housing and a buffer container housing operatively connectable to a reaction tube.

  The present invention relates to the collection and preparation of samples to be analyzed outdoors or in the field for the presence of biological warfare agents (eg, anthrax, wild boar, plague, or smallpox).

  Traditional PCR collection and preparation requires skills that exceed those of most field personnel. General PCR collection and preparation is a complex procedure.

  Conventional PCR collection and preparation requires processing the sample prior to analysis. The disadvantage of sampling and preparation is that it involves at least three complex steps. First, it removes inhibitors such as humic acid and metals in the environment. Second, the sample is filtered and concentrated. Third, the combination of reagents required for the PCR assay is added to the sample.

  Another disadvantage of conventional PCR collection and preparation is that the work is generally performed in the laboratory rather than outdoors. In general, such collection and preparation cannot be carried for field use for field inspection of biological warfare agents.

  In one embodiment of the present invention, an apparatus for collecting and preparing samples for PCR analysis is provided. The apparatus includes a buffer container housing and a plunger housing. The buffer container housing includes a buffer container disposed inside the buffer container housing. The plunger housing includes a plunger configured to be disposed inside the plunger housing. A swab is attached to one end of the plunger, and a reaction tube is attached to one end of the plunger housing. The buffer container housing is configured to be operatively connected to the plunger housing.

  In another aspect of the invention, a method for preparing PCR analysis is provided. The method includes providing an apparatus including a swab and a plunger and wiping the test surface with a swab to obtain a sample. The PCR analysis preparation method further includes inserting the swab into the buffer container by attaching the plunger chassis to the buffer container chassis using a torsional motion between the plunger housing and the buffer container housing, so that the buffer is released. Including allowing the swab to pass through and allowing the buffer and sample to proceed into the chamber for mixing with the reagent mixture. The method further includes whipping the device and thus feeding the prepared sample into the reaction tube.

  It should be understood that the foregoing general description and the following detailed description are exemplary only and are not restrictive of the claimed invention.

  These and other features, aspects, and advantages of the present invention will become apparent from the accompanying exemplary embodiments shown in the drawings, which are briefly described in the following description, appended claims, and brief description of the drawings. Will.

  Embodiments of the present invention will be described below with reference to the accompanying drawings.

  An apparatus 1 for collecting and preparing a sample for PCR analysis to determine the presence or absence of a biological agent (eg, anthrax, wild boar, plague, or smallpox) is disclosed in FIG. The apparatus 1 includes a buffer container housing 10 operatively connected to a plunger housing 60, a buffer container 20, a plunger 50, a swab 30, a swab support 40, and a reaction tube 70.

  As can be seen from FIGS. 2 (a) and 2 (b), the buffer container housing 10 includes a tab 14 at one end of the cylindrical housing portion 16. The cylindrical housing portion 16 includes an outwardly protruding thread 12. As can be seen from FIGS. 4A and 4B, the screw thread 12 is configured to correspond to the screw groove 62 of the plunger housing 60. Tab 14 is configured to assist the user in twisting or screwing buffer container housing 10 with plunger housing 60.

  The buffer container 20 may be any suitable container for containing a buffer, such as Whatman's Mini UniPrep brand buffer chamber 20.

  A swab or swabbing matrix 30 shown in FIG. 1 is attached to one end of the plunger 50. The swab support 40 supports the swab 30 on the plunger 50. As can be seen from FIGS. 3 (a) and 3 (b), the swab support 40 includes a convex framework 42 and a frame 44. The frame 44 can be substantially circular. The convex framework 42 is disposed inside the frame 44 and is configured to ensure that the swab 30 protrudes away from the plunger 50 and engages the surface on which the sample is taken for analysis. The

  The swab 30 is configured to sample a sample for PCR analysis that determines the presence or absence of a biological agent. The swab 30 can be formed in a dome shape.

  The swab 30 can be a filter-like material or any other material suitable for collecting the substance to be analyzed, such as IsoCode paper.

  As can be seen from FIGS. 4 (a), 4 (b), and 4 (c), the plunger housing 60 includes a thread groove 62 on the inner surface of the plunger housing 60. The plunger housing 60 can have a cylindrical shape. The radially extending tab 64 can project from the outer surface of the plunger housing 60. A radially extending tab 64 ensures that the reaction tube 70 properly engages a slot of a detector for further analysis of the sample (not shown), and (2) that the device is properly handled with a heavy hazardous material glove. Make sure you can.

  The plunger 50 shown in FIG. 1 can be placed inside the plunger housing 60 with the swab 30 and swab support 40 attached. For example, reagent beads (not shown), such as ceramic mixing beads, can be placed inside the interior chamber (not shown) of the plunger 50.

  Plunger 50 may include Whatman's Mini UniPrep brand plunger 50 or any other suitable plunger 50.

  As can be seen from FIGS. 5 (a), 5 (b), and 5 (c), the reaction tube 70 includes a tube portion 74 and a cup portion 72 at one end of the tube portion 74. An alignment tab 73 extends from the edge of the cup 72. The alignment tab 73 is configured to ensure that the reaction tube 70 is properly attached, aligned and held on the plunger housing 60.

  The reaction tube 70 can include any suitable tube for holding the prepared sample for further analysis, such as a Bio-Seq brand reaction tube 70. The reaction tube 70 can be disposable.

  The device 1 can be used to wipe surfaces that may be contaminated with biological warfare agents. The swab 30 of the device 1 attached to one end of the plunger 50 takes a sample of possible contaminants and then inserts it into the buffer container 20. The swab 30 is inserted into the buffer container 20 by a twisting operation between the plunger housing 60 and the buffer container housing 10. The speed at which the swab 30 is placed inside the buffer container 20 is controlled by a twisting action, and therefore it controls the speed at which the buffer contained within the buffer container 20 passes through the swab 30 and elutes or washes out the sample. The sample and buffer go into the internal chamber, thus rehydrating the lyophilized reagent beads, which are then mixed with the sample. The sample is then prepared for analysis and fed into the reaction tube 70. The sample advances into the reaction tube 70 by a whipping operation. The user of the apparatus 1 whips the apparatus 1 several times to feed the prepared sample into the reaction tube 70. The reaction tube 70 attached to the apparatus 1 can then be placed inside a hand-held detector for analyzing the sample.

  The device 1 can be configured to an appropriate size and weight so that it can be easily carried for use outdoors. For example, the device 1 can be carried by an operator's hand so that it can be easily carried.

  In an alternative embodiment, the buffer container 20 includes a membrane 22 (not shown) that covers one end of the buffer container 20. Plunger 50 and swab 30 break membrane 22 by a twisting action that pushes swab 30 into buffer container 20. When the membrane 22 breaks, a buffer (not shown) is released from the buffer container 20 and passes through the swab 30 to elute the sample.

  In the alternative embodiment shown in FIG. 6, a spike 125 is placed between the swab 130 and the buffer container 120. The spike 125 is configured to rupture the membrane 122 covering the buffer container 120 to release the buffer from the buffer container 120 and flow through the swab 130. The spike 125 ruptures the membrane 122 covering the buffer container 120 by pushing the spike 125 into the buffer container 120 by a twisting operation between the buffer container housing 110 and the plunger housing 160.

  In another alternative embodiment, the swab 30 can be covered by a mesh 32 (not shown). Alternatively, the swab 30 can cover the mesh 32.

The device can include additives, such as indicator chemical dyes, that improve the visibility of the combined sample-reagent mixture prior to introduction into the instrument. This dye can also be sensitized to reagent materials so that one color appears before rehydration of the reagent beads and the sample and another color appears after the sample is thoroughly mixed with the reagent. it can. Another feature of the dye is that, prior to PCR treatment, a color change can be used to indicate that the pH of the unknown sample is not compatible with the reagent system. In a preferred embodiment, Cresol Red, chemical formula C ₂₅ H ₁₈ O ₅ S is placed in the buffer. The normal color of this material in neutral solution is yellow. When properly mixed with PCR reagents, the solution turns purple to red. If for some reason no reagent is present, or if the sample has the wrong pH, the buffer will turn yellow indicating a problem with the reaction.

  Given the disclosure of the present invention, those skilled in the art will recognize that there are other embodiments and variations within the scope and spirit of the invention. Accordingly, all modifications that can be attained by a person skilled in the art from the present disclosure within the scope and spirit of the present invention shall be included as further embodiments of the present invention. The scope of the invention is to be defined as set forth in the claims.

1 is an exploded view of an embodiment of an apparatus for collecting and preparing a sample for PCR analysis. FIG. It is a perspective view of the buffer container housing | casing of FIG. It is a side view of the buffer container housing | casing of FIG. It is a perspective view of the swab support body of FIG. It is a side view of the swab support body of FIG. It is a perspective view of the plunger housing | casing of FIG. It is sectional drawing of the plunger housing | casing of FIG. It is a side view of the plunger housing | casing of FIG. It is a perspective view of the reaction tube of FIG. It is a side view of the reaction tube of FIG. It is sectional drawing of the reaction tube of FIG. FIG. 6 is an exploded view of another embodiment of an apparatus for collecting and preparing a sample for PCR analysis.

Claims (34)

An apparatus for collecting and preparing a sample for polymerase chain reaction analysis comprising:
A buffer container housing;
A buffer container;
A swab attached to one end of the plunger;
A reaction tube attached to the plunger housing,
The plunger is configured to be disposed within the plunger housing; and
The buffer container is disposed inside the buffer container housing, and the buffer container housing is operatively connected to the plunger housing;
apparatus.   The apparatus of claim 1, wherein the buffer container housing includes a tab at one end of the cylindrical housing portion.   The apparatus of claim 2, further comprising an outwardly projecting thread on the cylindrical housing portion.   The apparatus of claim 1, wherein the swab comprises a filter-like material.   The apparatus of claim 1, further comprising a swab support for supporting the swab.   The apparatus of claim 5, wherein the swab support includes a convex framework disposed inside a frame configured to position the swab such that the swab protrudes away from the plunger.   The apparatus of claim 1, further comprising a thread in an inner surface of the plunger housing.   The apparatus of claim 7, wherein the thread groove is configured to correspond to a thread projecting from a housing portion of the buffer container housing.   The apparatus of claim 1, further comprising at least one radially extending tab protruding from the plunger housing.   The apparatus of claim 1, wherein the reaction tube comprises a cup and a tube.   The apparatus of claim 10, further comprising at least one alignment tab disposed at an edge of the cup and configured to hold the reaction tube within the plunger housing.   The apparatus of claim 1, wherein the swab is inserted into the buffer container by a twisting motion between the plunger housing and the buffer container housing.   The buffer container housing and the plunger housing are configured such that a buffer contained in the buffer container passes through the swab when the swab is inserted into the buffer container. Equipment.   The apparatus of claim 1, further comprising at least one reagent bead disposed in the interior chamber of the plunger for mixing with the sample.   The apparatus of claim 1, wherein the reaction tube is configured to receive a sample of material prepared to undergo PCR analysis by whipping the apparatus.   The apparatus according to claim 1, wherein the buffer container housing, the plunger housing, and the reaction tube are configured to be portable.   The apparatus of claim 1, wherein the buffer container includes a membrane covering one end of the buffer container.   18. The apparatus of claim 17, wherein the membrane is ruptured by a twisting action that pushes the swab into the buffer container and the buffer is released by rupture of the membrane.   The apparatus of claim 1, further comprising a spike disposed between the buffer container and the swab, wherein the spike is configured to break a membrane covering one end of the buffer container.   The apparatus of claim 1, further comprising a mesh covering the swab.   The apparatus of claim 1, further comprising an additive to improve the visibility of the sample and reagent mixture prior to processing.   The apparatus of claim 1 further comprising an indicator dye.   23. The apparatus of claim 22, wherein the color of the dye changes when a reagent is present and properly mixed.   23. The apparatus of claim 22, wherein the color of the dye changes if the pH of the sample is correct.   23. The device of claim 22, wherein the dye comprises cresol red. An apparatus for holding a sample for polymerase chain reaction analysis comprising:
A swab holder, a buffer holder, and a container for collecting a sample, wherein the swab holder and the buffer holder are configured to be joined together, thereby placing the sample in the container Configured as
apparatus.   27. The apparatus of claim 26, wherein the container is tubular.   27. The apparatus of claim 26, wherein the container includes a funnel-shaped sample receptacle.   27. The apparatus of claim 26, wherein the container contains a reagent mixture.   27. The apparatus of claim 26, wherein the plunger includes at least one ceramic mixing bead.   27. The apparatus of claim 26, further comprising a housing covering the buffer holder and including a gripping portion.   27. The apparatus of claim 26, further comprising a swab at one end of the swab retainer.   34. The apparatus of claim 32, further comprising a mesh covering the swab. Providing a device comprising a swab and a plunger, wiping the test surface with the swab and taking a sample;
By attaching the plunger housing to the buffer container housing using a torsional motion between the plunger housing and the buffer container housing, the swab is inserted into the buffer container so that the buffer is released and passes through the swab. Allowing the buffer and sample to proceed into the chamber for mixing with the reagent mixture;
Whipping the device and thus feeding the prepared sample into the reaction tube;
A method for preparing polymerase chain reaction analysis, comprising:

Images