Method and device for preparing a sample of biological origin in order to determine at least one constituent contained therein

Abstract

The invention relates to a method of preparing an original sample ( 3 ) of biological origin with a view to detecting at least one component contained in it, whereby the original sample ( 3 ) is enclosed in a first container ( 1 ) and at least a part of this original sample ( 3 ) is transferred from the first container ( 1 ) into at least one reaction container, a reagent ( 9 ) or reagent mixture being placed beforehand in at least one of the reaction containers in order to prepare the original sample ( 3 ) and the component to be detected. In order to transfer at least a part of the original sample ( 3 ), two respective containers ( 1, 6 ) are connected to one another to form a closed, airtight system at least until at least the component(s) to be detected has (have) been rendered stable by reacting it (them) with the reagent ( 9 ) or reagent mixture at room temperature.

1 - 65 . (canceled) 
     
     
         66 . Device for establishing a closed, airtight flow connection between an interior of a first container and an interior of at least one other container, with a connecting region for the first container and at least one other connecting region for the at least one other container, a flow passage being disposed between the connecting regions, and with a respective piercing means for piercing a container seal in the connecting regions, wherein at least the connecting region ( 26 ) for the first container ( 1 ) has a retaining device for releasably securing the first container, by means of which a retaining force can be exerted on the first container ( 1 ) which is greater than the force counteracting separation of the other container ( 6 ) in the other connecting region ( 25 ) from the other connecting region ( 25 ). 
     
     
         67 . Device for establishing a closed, airtight flow connection between an interior of a first container and an interior of at least one other container, with a first connecting region for the first container and at least one other connecting region for the at least one other container, a flow passage being disposed between the connecting regions, and with a respective piercing means for piercing a container seal in the connecting regions, wherein a shut-off element ( 70 ) is disposed in the flow passage ( 38 ), by means of which the flow connection can be interrupted. 
     
     
         68 . Device as claimed in  claim 67 , wherein the shut-off element ( 70 ) has a rotatable passage which can be displaced to establish a flow connection with the flow passage ( 38 ). 
     
     
         69 . Device as claimed in  claim 66 , wherein the flow passage ( 38 ,  128 ,  154 ) is a double-ended cannula and the cannula ends serve as the means for piercing a container seal ( 115 ,  140 ), such as a septum ( 13 ,  28 ,  75 ,  87 ,  110 ,  146 ) for example. 
     
     
         70 . Device as claimed in  claim 66 , wherein a middle region ( 27 ) is provided between the connecting regions ( 25 ,  26 ) and the flow passage ( 38 ,  128 ,  154 ) is secured in this middle region ( 27 ) with the aid of one or more locking devices. 
     
     
         71 . Device as claimed in  claim 66 , wherein a cavity ( 43 ) is disposed between these two connecting regions ( 25 ,  26 ), through which the flow passage ( 38 ,  128 ,  154 ) is inserted, the volume of which is so dimensioned that at least a part of an airtight elastically deformable protective cap ( 46 ) secured to a part of the flow passage ( 38 ,  128 ,  154 ) projecting into the connecting region ( 26 ), in particular in the cavity ( 43 ), and made from a self-closing, elastic material which can be pierced, e.g. a valve bellows, can be accommodated by the cavity ( 43 ). 
     
     
         72 . Device as claimed in  claim 66 , wherein the connecting region(s) ( 25 ,  26 ) is or are at least partially provided in the form of an at least substantially cylindrical wall ( 31 ,  32 ), and an internal diameter of the cylindrical wall ( 31 ,  32 ) may be so dimensioned that a friction lock is created between an internal surface of the cylindrical wall ( 31 ,  32 ) and a container surface. 
     
     
         73 . Device as claimed in  claim 72 , wherein a catch mechanism is provided on the internal face of the cylindrical wall ( 31 ,  32 ) for receiving a part of the container ( 1 ,  6 ,  83 ,  98 ,  136 ), in particular for a part of a cap of a blood sample tube. 
     
     
         74 . Device as claimed in  claim 72 , wherein the cylindrical wall ( 31 ,  32 ) is made from a deformable material, which conforms to the external contours of the inserted container ( 1 ,  6 ,  83 ,  98 ,  136 ) when inserted. 
     
     
         75 . Device as claimed in  claim 66 , wherein a cross-sectional tapered region ( 42 ) is disposed in the middle region ( 27 ) between the first and the second connecting region ( 25 ,  26 ). 
     
     
         76 . Device as claimed in  claim 75 , wherein an external device surface in the region of the cross-sectional tapered region ( 42 ) is provided with knurling. 
     
     
         77 . Device as claimed in  claim 66 , wherein a sealing device, e.g. a sealing ring, is disposed on the device surface in the second connecting region ( 26 ) for the second container ( 6 ,  83 ,  98 ,  136 ). 
     
     
         78 . Device as claimed in  claim 66 , wherein the flow passage ( 38 ) is retained so that it can slide. 
     
     
         79 . Device as claimed in  claim 66 , wherein the flow passage ( 38 ,  128 ,  154 ) is made up of two parts and a flow passage part ( 61 ,  62 ) co-operates with each connecting region ( 25 ,  26 ). 
     
     
         80 . Device as claimed in  claim 79 , wherein a spring element ( 63 ) is disposed between the two flow passage parts ( 61 ,  62 ). 
     
     
         81 . Device as claimed in  claim 80 , wherein the spring force of the spring element ( 63 ) is greater than the force needed to pierce the container seal ( 115 ,  140 ), in particular the septum ( 13 ,  28 ,  75 ,  87 ,  110 ,  146 ). 
     
     
         82 . Device as claimed in  claim 80 , wherein a releasable locking device ( 64 ) co-operates with the spring element ( 63 ), which holds the spring element ( 63 ) in a biased state. 
     
     
         83 . Device as claimed in  claim 66 , wherein it is made up of at least two parts which are joined to one another by means of a releasable connection. 
     
     
         84 . Device as claimed in  claim 83 , wherein the releasable connection is a screw connection, a bayonet fitting, a friction-locking connection or similar. 
     
     
         85 . Device as claimed in  claim 66 , wherein the flow passage ( 38 ,  128 ,  154 ) has several, in particular mutually inter-connecting, part-flow passages ( 112 ) at least in the region of a connecting region ( 25 ,  26 ) and of a device for piercing a container seal, and these part-flow passages ( 112 ) serve as the device for piercing a container seal. 
     
     
         86 . Container with an interior which is at least partially enclosed by a container base and a container wall, the container wall having an inlet opening which is preferably disposed opposite the container base in the direction of a container longitudinal mid-axis, wherein the inlet opening ( 123 ) is provided in the form of a device ( 125 ) for establishing a closed, airtight connection to another container or the device fits into or on the inlet opening ( 123 ), this device ( 125 ) having a connecting region ( 132 ) for releasably attaching another container, such as a screw connection, a bayonet fitting, in which a means for piercing a container seal of the other container is disposed. 
     
     
         87 . Container as claimed in  claim 86 , wherein at least one other opening is provided in the container base ( 120 ) and/or in the container wall ( 121 ) and is designed as a device ( 125 ) for establishing an airtight connection to another container or the device ( 125 ) is disposed in or on the opening. 
     
     
         88 . Container as claimed in  claim 86 , wherein the container base ( 120 ) and/or the container wall ( 121 ) is of an elastically deformable design. 
     
     
         89 . Container as claimed in  claim 86 , wherein at least a part of the container base ( 120 ) is designed as a liquid-tight, sealing plunger. 
     
     
         90 . Container as claimed in  claim 86 , wherein a reagent ( 9 ) or reagent mixture is disposed in its interior, such as a lysing buffer, an eluting fluid, for example. 
     
     
         91 . Container as claimed in  claim 86 , wherein at least one dividing wall is disposed in the interior. 
     
     
         92 . Container as claimed in  claim 86 , wherein a displaceable plunger is disposed in at least one of the part-volumes created by the dividing wall, the maximum cross section of which matches the cross section of the part-volume. 
     
     
         93 . Container as claimed in  claim 89 , wherein the plunger can be locked in at least one of its end positions. 
     
     
         94 . Container as claimed in  claim 89 , wherein a hollow plunger rod incorporating a chamber is disposed on the side of the plunger remote from the part-volume. 
     
     
         95 . Container as claimed in  claim 94 , wherein a receptacle containing a matrix is disposed in the chamber. 
     
     
         96 . Container as claimed in  claim 95 , wherein the matrix is intended as a means of separating a component from at least a part of the original sample. 
     
     
         97 . Container as claimed in  claim 95 , wherein the matrix is disposed in a receptacle incorporating an outlet. 
     
     
         98 . Container as claimed in  claim 97 , wherein the outlet is fitted with a barrier device, which can be moved from a position in which it seals the outlet to a position in which it releases it. 
     
     
         99 . Container as claimed in  claim 92 , wherein a device is disposed in an end region of the plunger rod remote from the part-volume in order to receive another container. 
     
     
         100 . Container as claimed in  claim 92 , wherein a reagent ( 9 ) or reagent mixture is placed in a second part-volume beforehand. 
     
     
         101 . Container as claimed in  claim 92 , wherein the two part-volumes are linked to one another via a transfer passage. 
     
     
         102 . Container as claimed in  claim 101 , wherein the transfer passage can be closed off by at least one barrier mechanism. 
     
     
         103 . Container as claimed in  claim 92 , wherein at least one part-volume has an opening which is sealed by a septum. 
     
     
         104 . Container as claimed in  claim 86 , wherein at least one membrane, e.g. an air bellows, is disposed in the interior and is provided with a connecting piece for a supply line via an airtight valve system which can be shut off. 
     
     
         105 . Container as claimed in  claim 86 , wherein the flow passage ( 128 ) is a hollow needle. 
     
     
         106 . Container as claimed in  claim 86 , wherein a cavity is disposed below the connecting region ( 132 ) in the direction towards the interior through which the flow passage ( 128 ) is inserted and its volume is so dimensioned that at least a part of a protective cap ( 129 ) is accommodated by this flow passage ( 128 ), the former being secured in the latter in particular. 
     
     
         107 . Container as claimed in  claim 86 , wherein the connecting region ( 132 ) is an open cylinder with a wall. 
     
     
         108 . Container as claimed in  claim 107 , wherein the internal diameter of the cylinder is such that a friction lock is created between an internal surface of its wall and the other container. 
     
     
         109 . Container as claimed in  claim 107 , wherein a catch mechanism is provided on the internal surface of the wall of the cylinder, in particular for the cap of a blood sample tube. 
     
     
         110 . Container as claimed in  claim 107 , wherein the wall of the cylinder is made from a material which conforms to the external contours of the other container when the latter is inserted. 
     
     
         111 . Container as claimed in  claim 86 , wherein the flow passage ( 128 ) is retained so that it can slide. 
     
     
         112 . Device for cutting a sample of biological origin into smaller pieces, comprising a container with an interior which is at least partially bounded by a container wall and a container base, wherein at least one cutting device ( 142 ), e.g. a blade, is disposed on a surface of the container wall ( 138 ) and/or the container base facing the interior ( 137 ). 
     
     
         113 . Device for cutting a sample of biological origin into smaller pieces, incorporating a sealing device for a container, such as a blood sample vessel, a centrifuge vessel, for example, with a sealing body with and end region facing the container to be sealed, wherein a cutting device ( 142 ) is disposed in the region of the end region and a drive shaft ( 148 ) of the cutting device ( 142 ) is inserted through the sealing body in order to establish a connection to a drive unit ( 150 ), such as a motor for example. 
     
     
         114 . Device as claimed in  claim 113 , wherein the sealing body is provided at least partially in the form of a self-closing septum ( 146 ). 
     
     
         115 . Device as claimed in  claim 113 , wherein the drive shaft ( 148 ) is a tube and the end region lying opposite the cutting device ( 142 ) may optionally be designed to pierce a septum. 
     
     
         116 . Device as claimed in  claim 113 , wherein the sealing body is retained by a screw cap. 
     
     
         117 . Analysis kit consisting of at least a first container enclosing a first interior which is designed as the sample vessel and is designed in particular to contain samples of biological origin and biological matrices, such as blood and phases separated therefrom for example, another container enclosing a second interior in which at least one reagent is placed, such as a lysing buffer for cells such as blood cells for example, a stabilising buffer for nucleic acids and proteins, a releasing buffer, a preserving buffer and the solid salts of these buffers, and, optionally, a device for connecting the first interior to the second interior, wherein at least one container is of the type as recited above and the device for connecting the interiors is of the type as claimed in  claim 66 . 
     
     
         118 . Analysis kit as claimed in  claim 117 , wherein it incorporates a third container enclosing a matrix ( 15 ) to which sample constituents such as nucleic acids, proteins, are bonded, in particular by adsorption. 
     
     
         119 . Buffer for preserving and/or releasing at least one component from an original sample of biological origin, e.g. a nucleic acid, a protein, containing ammonium sulphate in a quantity within a range having a lower limit of 10% w/v, preferably 25% w/v, in particular 30% w/v, and an upper limit of 100% w/v, preferably 80% w/v, in particular 60% w/v, a chaotropic salt in a concentration selected from a range with a lower limit of 1 mol/l, preferably 2 mol/l, in particular 2.5 mol/l, and an upper limit of 6 mol/l, preferably 5 mol/l, in particular 3.5 mol/l, and a buffer substance in a concentration selected from a range with a lower limit of 0.1 mol/l, preferably 0.2 mol/l, in particular 0.25 mol/l and an upper limit of 0.5 mol/l, preferably 0.4 mol/l, in particular 0.3 mol/l, and optionally a detergent and/or a complexing agent. 
     
     
         120 . Use of the device as claimed in  claim 66 , for analyzing samples of biological origin or containing biological matrices. 
     
     
         121 . Use of the device as claimed in  claim 66 , for detecting nucleic acid. 
     
     
         122 . Use of the container as claimed in  claim 86 , for analyzing samples of biological origin or containing biological matrices. 
     
     
         123 . Use of the container as claimed in  claim 86 , to detect nucleic acid. 
     
     
         124 . Use of the device as claimed in  claim 112 , to cut tissue samples into smaller pieces. 
     
     
         125 . Use of the analysis kit as claimed in  claim 117 , for analyzing samples of biological origin or containing biological matrices.