AT414209B - COLLECTION TANK FOR LIQUIDS - Google Patents

Description

2

AT 414 209 B

The invention relates to a collection vessel for liquids for use in nucleic acid analysis, e.g. Blood collection tube, with one of a vessel shell and optionally a vessel bottom, which (r) a vessel inner wall form (t), at least partially surrounded vessel interior through at least one vessel opening, which is closable with a septum 5 provided with a closure device, accessible, a carrier for reagents or reagent mixtures for use in a collecting vessel, with a carrier body with a carrier body surface, and an analysis kit for analyzing liquids of biological origin with at least one collecting vessel for receiving the liquid or components of the liquid and at least one carrier for receiving a reagent or io reagent mixture.

In the field of laboratory diagnostics, the past is characterized by making analytical methods or analyzers for the examination of fluids of biological origin, such as blood, more and more efficient, not least in order to lower the detection limits for certain blood components. However, it is still necessary for the analysis of complex blood components, such as genetic information, i. Desoxy and ribonucleic acid (DNA, RNA), a variety of different preparation steps is required. One reason for this is that the DNA or RNA are not or only to a limited extent stable and thus already from the time of blood sampling undergo a continuous degradation process, triggered by nucleases, enzymes or the like., Which are contained in cells, and Thus, the result of the analysis is falsified by unwanted fission products. Reproduction of physiological control, which does not affect the action of cellular and extracellular nucleases as long as the cells are in their normal environment, has not been successful. The removal of blood leads to more or less 25 changes in the nucleic acids contained in the cells, so that just a long-term storage of blood leads to aging or destruction of the cell. Along with this, naturally problems also arise in the investigation of viruses contained in the blood via their genetic coding. One of today's usual ways to store blood unchanged over a longer period, is the lowering of the storage temperature, for example with the help of liquid nitrogen. Another possibility is that certain additives are added to the blood or, more generally, to a biological fluid. If possible, these additives should be selected so as not to interfere with subsequent analysis. For example, from EP 0 818 542 A1 a process is known with the aid of which the nucleases are to be inhibited by the addition of a guanidinium salt in powder form.

DE 195 19 886 A1 describes a blood collection device, which can be closed by a puncturable plug. This plug is designed so that in a Ausspa-40 tion protrudes a vessel with an additive contained therein, this vessel also has a puncturable, but not resealable bottom. A cannula penetrating the stopper penetrates into the vessel with the additive, and through the further advancement of the cannula, its bottom is pierced. This makes it possible to bring the additive into contact with the blood already during blood sampling. By means of this technique, the disadvantage resulting from the use of polyethylene terephthalate tubes, namely the water vapor permeability and thus the impairment of the additive present in the blood collection device, for example by lump formation, should be eliminated.

A disadvantage both in the method according to EP-A1 and in the blood collection device according to DE-A1 is that, in principle, only a single method step in the chain of preparation steps necessary for the analysis of blood constituents is covered, namely in the case of EP -A1 the stabilization of the nucleic acid and in the case of DE-A1 the addition of a blood coagulation preventing or a blood coagulation accelerating additive. In addition, the DE-A1 has the disadvantage that no defined addition of the additive 55 takes place, as always a non-predetermined residue remains in the vessel. 3

AT 414 209 B

EP 0 875 202 A2 shows another way of introducing an additive into a blood sample or into a part of a blood sample. For this purpose, this blood collection tube in the lower part of a matrix of fiber material, which is impregnated with a solution of an aqueous additive. To protect the additive, this blood collection tube further contains a water-immiscible liquid which has a specific gravity greater than 1.07, and thus the aqueous additive can be encapsulated. In the blood collection tube, a device is also included, which allows a clean separation of the phases after filling the tube and centrifuging. With the help of this system, a coagulation reagent should be added to the blood sample and, in turn, only a single step of the preparatory phase for the detection of individual components of the blood can be covered.

US 4,703,763 A1 describes a syringe for blood sampling, which is specially designed to obtain blood samples for subsequent blood gas analysis. In particular, error rates during sampling by contaminating the sample with air are verrin-15 siege.

For this purpose, the syringe in the interior of an air-permeable and liquid-impermeable closure device with a channel, which is closed for example by a self-closing, porous polyethylene. Upon contact of the blood with the polyethylene, this is dissolved in the aqueous phase of the blood, so that it is thickened and the channel in the closure element is liquid-impermeable.

The inner surface of the syringe body is preferably coated with heperin to prevent coagulation of the blood. Contact between the heperin and the blood takes place during the blood draw.

From US 4,639,419 A1 there is provided a test tube for the immunological analysis of body fluids, e.g. Blood, serum, plasma, etc., known. 30 The test is designed as a staining test. For this purpose, two separate, colored reagents are presented in the test tube. The first consists of a carrier to which a dye is attached and which is coated with an antibody or antiserum to obtain the specific antigen-antibody reaction of a positive assay. The second reagent also consists of a colored carrier, the color being different from that of the first reagent, and this second carrier is a serum protein or an inert molecule or colored carrier particles coated in such a way that the antigen-antibody Reaction not done, formed.

After filling the test tube with the liquids to be analyzed, the two reagents are dissolved or suspended in this liquid and subsequently a mixed color, if the test is negative. If the result is positive, the specific antigen-antibody reaction takes place and the supernatant fluid has the color of the second reagent. The invention has for its object to provide a way to simplify the analysis of liquids of biological origin or biological matrices.

This object of the invention is achieved in that a lysis reagent is introduced into the vessel interior of the collecting vessel and at least one reagent or reagent mixture with which 50 components of the liquid disrupted by the lysis are separated or immobilized and / or stabilized is arranged on a carrier. The advantage here is that the user now has a collecting vessel, e.g. can be made available for blood sampling, which can be used not only for receiving the liquid, but at the same time preferably with the introduction of the liquid into this collecting vessel an application of the liquid liquid or fission products of this liquid and optionally one at least 4

AT 414 209 B partial analysis of individual components of this liquid takes place. It can thus be used e.g. the blood loss, in particular the time to the presence of an analysis result, be shortened and, if appropriate, be carried out by the user without the intervention of a laboratory. In addition, an opening of the 5 cells is already possible during the filling process of the collection vessel.

An advantage of this is also an embodiment according to claim 2, since thus the genetic information is available faster and, if necessary, nucleic acids can be stored over a longer period in the matrix, as is advantageous, for example, for the elucidation of io criminal cases.

It is further an embodiment according to claim 3 of advantage, since thus an indirect, qualitative statement about the presence or absence of nucleic acids or nucleic acid mixtures can be made. 15

However, it is also an embodiment according to claim 4 advantageous, since thus a binding of the substance to be analyzed on a solid phase is possible and thus the latter can be separated from the matrix together with the substance to be analyzed. Furthermore, it is in this embodiment of advantage that thus a possibility can be created, 20 can be provided with the reagents over a longer period of time depending on dissolution behavior in the liquid.

With the development according to claim 5, the advantage can be achieved that the collected liquids can be mixed without much effort with the reagent or reagent mixture 25.

By forming the reagent or reagent mixture according to claim 6, a division of the components of the liquid 2 to a plurality of phases and thus a simpler separation of the substances to be examined can be achieved. 30

Another advantage is an embodiment of the reagent or reagent mixture according to claim 7, since the use of at least one of these chaotropic salts, the study of genetic material, in particular the stability of nucleic acids in the matrix, can be improved. 35

However, a further development according to claim 8 is also advantageous, since it can effectively provide a collecting vessel with a reagent or reagent mixture of specific concentration for carrying out a separation according to density gradients. A further advantage is an embodiment according to claim 9, as it allows the analysis of hydrophobic substances can be facilitated.

By continuing according to claim 10, it is possible to design the reagent or reagent mixture so that this can be used both for analysis and for the physical separation of the constituents of the liquid 45.

According to the embodiment of claim 11, it is possible to separate the carrier either together with the substance to be analyzed, or with the matrix from the rest of the liquid. 50

By the embodiments according to claims 12 and 13, an advantageous surface enlargement of the carrier can be achieved.

According to the embodiment according to claim 14, a separation by molecular weight and thus a corresponding fractionation of the substances to be analyzed is possible. 5 AT 414 209 ß

Due to the development according to claim 15, the carrier can be adapted to a wide variety of requirements in a simple manner.

Another advantage, however, is a further development according to claim 16, since this allows a simple separation of the carrier from the matrix.

The design of the collecting vessel according to claim 17, any liquids can be divided into several phases and separated, regardless of whether it is clear before using the collection vessel, whether the substance to be analyzed is present in the heavier or lighter phase io.

But also advantageous is a development according to claim 18, whereby a separation of a medium-heavy phase of a light and a heavy phase is possible without the latter must be removed from the vessel. But it is also possible an embodiment according to claim 19, since it allows the division of the components of the liquid to be examined in several phases using laboratory-standard aids. It is further possible a variant embodiment according to claim 20, is possible with the individual components of the liquid to be analyzed selectively from the matrix without the aid of other auxiliaries.

However, the object of the invention is also achieved by a carrier for reagents or reagent mixtures for use in a collecting vessel in which a reagent or reagent mixture is disposed on the carrier body, with which at least individual components of a liquid of biological origin are separated or immobilized and / or stabilized. The advantage here is that both the carrier and the reagent or reagent mixture can be easily adapted to the problem asked and also such carrier universal 30 can be used in any vessel. In particular, by this embodiment of the carrier this can be separated in a simple manner from the liquid to be analyzed, for example with the aid of mechanical means.

It is advantageous if according to claim 22, the carrier body surface is coated with the reagent or reagent mixture, since thus a subsequent separation of the spent reagent or reagent mixture or the adherent substance to be analyzed by the carrier is possible and thus the latter under circumstances appropriate preparation can be used again. In this case, embodiments according to claims 23 to 25 are advantageous, whereby a correspondingly increased surface area can be made available for the reaction of the substance to be analyzed.

By continuing according to claim 26 of the carrier according to the invention can be introduced in a simple manner in a liquid to be analyzed and removed from it.

Another advantage is an embodiment according to claim 27, whereby a selective application of polarizable or polar substances is possible. 50

In this case, the embodiments according to claims 28 and 29, a certain cohesion of the individual particles and thus their easier removal from the liquid to be analyzed allows, at the same time this carrier, for example. can fill the entire cross-section of a receiving vessel without the passage of the liquid is impaired too much, at the same time achieved an increase in surface area 6

AT 414 209 B can be.

It is also advantageous embodiment according to claim 30, which in particular an adaptation of the available carrier body surface of elongated Analysge-5 vessels is possible. Furthermore, the carrier can be provided with a closure device, so that the carrier can simultaneously fulfill the closure function after insertion into a corresponding vessel.

In this case, a development according to claim 31 is advantageous, according to which a tight closure of the corresponding receptacle can be achieved and thus intimate mixing of the liquid to be analyzed with the carrier or the components of the carrier without spilling the liquid is possible.

It is further an embodiment according to claim 32 possible because it can be achieved 15 that a closed container with such a carrier can be filled without this carrier must be removed from the vessel.

But are also advantageous embodiments according to claims 33 and 34, as it allows the supply of other reagents or washing liquids by the carrier in a simple way 20 and manner.

But also advantageous is a development according to claim 35, since thus an adaptation of the carrier to any cross-sections is possible. By the embodiment according to claim 36, an adaptation of the carrier to a variety of applications can be realized in a simple way.

The object of the invention is further solved by the analysis kit, in which the collecting vessel and / or the carrier is designed according to the invention / are. It is advantageous that the An-30 wender a coordinated system for the analysis of liquids can be provided, with a rapid investigation while reducing the necessary steps can be made possible.

By arranging further collecting vessels according to claim 38, the advantage 35 can be achieved that within a closed system several steps for the analysis of liquids can be performed and in this way it is also possible to achieve a simple separation of components of the liquids.

In this case, an embodiment according to claim 39 is advantageous because after separation of several-40 rer collection vessels from each other, the individual collection vessels are automatically sealed.

By assembling the analysis kit according to claims 40 and 41 further simplification of the analysis can be achieved. 45

The invention will be explained in more detail with reference to the embodiments illustrated in the drawings, wherein all representations are made greatly simplified.

Show: 50

1 shows an inventive collecting vessel with applied to the inner wall coating a reactive component.

FIG. 2 shows a collecting vessel according to the invention with an inserted carrier provided with a reactive component; FIG. 55 Fig. 3 shows another embodiment of the collecting vessel according to the invention with a 5 5 7

AT 414 209 B set carrier;

4 shows a collecting vessel according to the invention with coated carrier particles arranged in the collecting vessel interior;

5 shows an arrangement variant of the carrier particles in the collecting vessel;

FIG. 6 shows a carrier for introducing the reactive component into the collecting vessel; FIG.

Fig. 7 shows another embodiment of the carrier according to the invention;

8 shows a collecting vessel according to the invention with liquid reactive component;

9 shows a collecting vessel with a carrier for the reactive component which can be screwed onto it; 10 15 20 25

10 shows an analysis kit consisting of a collecting vessel and several mounted carriers;

11 shows an analysis kit consisting of a plurality of collecting vessels communicating with one another via cannulas;

12 shows a collecting vessel according to the invention with inserted separating device for better phase separation during or after centrifuging;

13 shows a collecting vessel according to the invention with a cap in which a carrier with the reactive component is contained;

Fig. 14 shows the carrier for the reactive component as part of a cannula;

15 shows a collecting vessel with carrier for the reactive component arranged on a closure device for the collecting vessel;

Fig. 16 is a branch vessel system with a reactive component coating attached to the inner vessel;

17 shows a further embodiment of the collecting vessel according to the invention in a simplified, schematic representation;

FIG. 18 shows an embodiment variant of the collecting vessel for multi-component analysis; FIG.

19 shows the closure device for the collecting vessel according to FIG. 18 in a schematically simplified plan view.

By way of introduction, it should be noted that in the differently described embodiments, 30 identical parts are provided with the same reference numerals and the same component names, wherein the disclosures contained in the entire description can be mutatis mutandis transferred to like parts with the same reference numerals or component names. Also, the location information chosen in the description, such as top, bottom, side, etc. related to the immediately described and illustrated figure and are to be transferred at 35 a change in position mutatis mutandis to the new situation. Furthermore, individual features or combinations of features from the different exemplary embodiments shown and described can also represent independent, inventive or inventive solutions. 40 In FIG. 1, a collecting vessel 1 according to the invention for liquids 2 of biological origin or biological matrices is shown schematically in section. A vessel interior 3 is accessible through at least one vessel opening 4. The vessel interior 3 is bounded by a vessel inner wall 5, which of a vessel shell 6 with two with respect to a Gefäßmantelmittelachse 7 opposite each other arranged Gefäßmantelstirnflächen 45 8, 9, where appropriate, as shown in FIG. 1, with the Gefäßmantelstirnfläche 9 a Ge fäßboden 10 may be integrally connected, and at least one, the vessel opening 4 occlusive closure device 11 is formed. This collection vessel 1 may be formed, for example, as known from the prior art blood collection tube. However, it is also possible to use the collecting vessel 1 for receiving other liquids 2. 50

The collecting vessel 1 may for example be made of a preferably thermoplastic material, e.g. Polyethylene terephthalate (PET), polypropylene (PP), polyvinyl chloride (PVC), polystyrene (PS) or the like., Glass, metal, ceramic or the like., Be formed or can also form several of these materials, the receptacle 1. 55 8

AT 414 209 B

The closure device 11 is preferably designed so that a perforable septum 12 is held therein. For this purpose, the closure device 11 may have an outer casing 13, which in turn may be made of plastic. The closure device 11 may e.g. be designed as a screw cap, plug cap or the like., And to allow a perforation of the septum 5 12, in the plane parallel to the Gefäßmantelstimflächen 8 a bore 14 have. Moreover, the closure device 11 may correspond to the known from the prior art closure devices for example blood collection tube.

It is advantageous if the outer casing 13 in the direction parallel to the vessel casing central axis 7 io the vessel shell 6 overlaps at least partially, as thus the risk of accidental ejection of the liquid 2 can be largely prevented when you open the filled receptacle 1.

The attachment of the closure device 11 to the collection vessel 1, i. on the vessel jacket 6, 15 can e.g. via a arranged on an outer jacket inner side 15 of the closure device 11, in Fig. 1 indicated by dashed lines thread 16. For this purpose, a corresponding web 17, for example, in the region of the vessel shell end face 8, be arranged on the vessel shell 6, so that engagement of this web 17 in the thread 16 is possible or it is also possible to provide a Gefäßmantelaußenwand 18 with a corresponding thread. 20

It is also possible, in particular when the closure device 11 is formed as a plug-in closure, that the attachment of the closure device 11 takes place solely by adhesive forces between the septum 17 and the vessel inner wall 5. Of course, these adhesive forces can assist in the variant with the thread 16. 25

Of course, it is also possible that the closure device 11 is formed solely by the septum 12 or are other closure devices 11, for example, have no bore 14, also usable. The material used for the septum 12 is above all perforable materials, e.g. Elastomers, for example rubber or the like, into consideration. This septum 12 is intended to fulfill the task that the perforation, which is caused by the piercing of the septum 12 with a cannula 19 or a corresponding other suitable device, after the withdrawal of the cannula 19 is again closed so that leakage of the liquid 2 is prevented from the 35 collection vessel 1. It should thus be avoided as well as by the aforementioned embodiment of the closure device 11, a contamination of the user of the collecting vessel 1, for example, with bacteria, viruses or the like., Which may be contained in the liquid 2. Preferably, in the vessel interior 3, a pressure greater than the standard pressure, i. 1013 mbar at 298.15 K, lower pressure prevailing. This can be achieved that the liquid 2 is spent by this pressure gradient automatically when needed in the vessel interior 3.

The pressure in the interior can be adjusted so that a predefined liquid volume 45 is introduced into the vessel interior 3.

The vessel bottom 10 can, as shown in FIG. 1, be designed semicircular or it is also possible to choose other embodiments. According to the invention, at least one reagent or reagent mixture 20 accessible through the vessel opening 4 is arranged in the vessel interior 3, with which at least individual constituents of the liquid 2 or the biological matrix or cleavage products of these constituents can be separated off and / or immobilized and / or stabilized. The arrangement may be such that the vessel inner wall 5 is coated with this reagent or reagent mixture 20 55. This coating can be made by any conventional, suitable techniques, 9

AT 414 209 B, which are known from the prior art, can be applied, for example it is possible to introduce a suspension of the reagent or reagent mixture 20 into a corresponding solvent, which should have a high vapor pressure, into the collecting vessel 1, whereupon Excess suspension is removed again and the solvent is evaporated to 5 drying of the reagent or reagent mixture 20, optionally with an increase in temperature, so that in the end the reagent or reagent mixture 20 is present in the solid state.

Of course, known methods of the spray technique for coating the io collecting vessel 1 with the suspension or the reagent or reagent mixture 20 can be used.

The reagent or reagent mixture 20 may for example be selected or composed so that it is selective for nucleic acids or nucleic acid mixtures. In this case, the selectivity can be designed both for individual nucleic acids, for example deoxyribonucleic acids (DNA) or ribonucleic acids (RNA). In this way, it is possible to bind nucleic acids of different origin to this reagent or reagent mixture 20, so that in the sequence after removal of the remaining liquid matrix these nucleic acids from the reagent or reagent mixture 20 eluted or the resulting reaction products 20 can be processed and so directly available for further analysis. Thus, the advantage can be achieved that essentially the substance to be analyzed or the substance mixture can be prepared so far with only one step that their detection either directly or with the interposition of only a few further preparation steps is possible (eg elution of the analyzed Substance). For this purpose, the reagent or reagent mixture 20 should be selected or assembled in such a way that stabilization and / or lysing takes place in the case of examination of nucleic acids in the blood during or shortly after blood collection and the subsequent immobilization of the nucleic acids takes place. 30 Lysing is necessary because blood usually consists of a mixture of cells, proteins, metabolites, etc. The cells in turn consist of erythrocytes, platelets and leukocytes, only the latter containing the nucleic acids. Thus, to deliver these nucleic acids to a possible reaction, it is necessary to lyse the cells. For qualitative analysis, it is further possible for the reagent or reagent mixture to contain a substance which reacts selectively for a particular nucleic acid, for example in order to make the nucleic acid accessible to subsequent fluorescence detection.

There is also the possibility that with the aid of the reagent or reagent mixture 20 a controlled degradation of the nucleic acid takes place and that the reagent or reagent mixture 20 selectively on these nucleic acid components, for. Mononucleotides, oligonucleotides or the like. Reacted. Furthermore, it is possible that, in the case where the liquid 2 contains components which react specifically with the particular nucleic acids, the reagent or reagent mixture 20 is selectively selected for these substances. Such nucleic acid degradation-ending ingredients may be e.g. Proteins, metabolites, nucleases, enzymes or the like.

In a modification of this embodiment, it is possible to use materials for the collecting vessel 1, which at least partially form the reagent or reagent mixture 20. For example, it has long been known that glass surfaces can selectively bind nucleic acids. This principle is also applicable to plastics, if they are modified accordingly. Thus, it is possible, for example, to use polymers which have terminal, charged or by appropriate pre-preparation, for example by acid treatments, loadable groups. By way of example, amino groups or hydroxyl groups may be mentioned for this purpose, in which amino groups, e.g. Diethylaminogruppen, by an appropriate the acid treatment step, the nitrogen can be positively charged and thus the Anbin- 10

For example, AT 414 209 B fertilizes the DNA via the phosphate group, i. In particular, an oxygen of this phosphate group is possible. Likewise, of course, nitrile polymers or copolymers or the like. Can be used, and it must always be ensured that these polymers have corresponding polarizable or polarized, preferably unbranched substituents 5. With the aid of this embodiment of the collecting vessel 1, it is possible in a simple manner, if an additional reagent or reagent mixture 20 is present in the collecting vessel 1, which takes the lysis and / or stabilization of blood samples, in particular nucleic acids, preferably quantitatively separate them. Since these are selectively held by the wall of the collection vessel 1, it is also possible to pour off the excess portion of the blood sample, so that the collection vessel 1 can be used in the further sequence for the washing and further processing of the nucleic acids.

Another example of such a variant of the collection vessel 1 is the use of a plastic with carboxy groups, which allows the attachment of Olygonucleoti-15. The corresponding lysis of the cells is of course also required in this embodiment.

In Fig. 2, another embodiment of the collecting vessel 1 according to the invention is shown. In contrast to the preceding embodiment variant, the reagent or reagent mixture 20 is not arranged as a coating on the vessel inner wall 5 but is located on a separate carrier 21.

This carrier 21 may be connected to the vessel 1, for example, by bonding with the vessel inner wall 5, or held by means of static friction at the desired position in Sam-25 melgefäß 1. It is also possible that, as indicated by dashed lines in Fig. 2, the collection vessel 1 on the vessel inner wall 5 has a protruding into the vessel interior 3 web 22, said web may be circumferentially formed over the entire circumference or even only partially over the scope of the collecting vessel 1 extends. As a circumference is the scope of the vessel inner wall 5 perpendicular to the Gefäßmantelmit-30 telachse 7 to understand. The latter embodiment has the advantage that the carrier 21 can be removed from the collecting vessel 1 in a simple manner and thus is available for further processing for the analysis of the respective substance to be detected or after removal of this substance for reuse. The carrier 21 may be formed, for example, in the form of a preferably inert filter. Thus, it is possible to use the carrier 21 made of glass, e.g. Glass fibers, silicate materials, cellulose, latex, etc. to manufacture.

In Fig. 3, another embodiment for the arrangement of the carrier 21 in the collection-40 vessel 1 is shown. In this case, the vessel shell 6 in the direction of the vessel jacket central axis 7, a dimension change 23, which is designed so that the diameter of the collecting vessel 1 is reduced in the direction of the vessel bottom 10. The resulting constriction is carried out in one stage according to the example of FIG. 3, but it is also possible that this dimensional change 23 takes place continuously and thus the diameter of the Ge-45 fäßöffnung 4 in the direction of the vessel bottom 10 decreases continuously. With the help of this dimensional change 23, it is possible to support the carrier 21, which has a predeterminable diameter, measured from the vessel bottom 10 in the direction of the vessel opening 4 at a certain height 24 and are therefore no further measures for holding the carrier 21 in the collecting vessel 1 needed. It can thus be achieved that the carrier 21 only so rests on the dimensional change 23 and thus a simple removal of the carrier 21 from the collecting vessel 1 is possible, for example by means of tweezers.

As shown in Fig. 3 further by the dashed or dot-dash line within the carrier 21, it is possible to divide the carrier 21 zonar and in these zones the Rea-gene gene or reagent mixture 20 composed differently, so that in the sequence 1 1

AT 414 209 B

Multi-component analysis can be done.

Likewise, it is of course possible to stack in this way a plurality of carriers 21 on top of each other, so that a separation of the individual carriers is possible in a simple manner and can be separated in sequence 5, the substances to be detected without the need for further separation steps.

If necessary, of course, the substance to be detected, such as DNA, RNA can be eluted from the carrier with the aid of an appropriate solvent and it is subsequently possible io further use by regeneration of the carrier.

The carrier 21 may have capillaries in its interior, for example in the manner of a filter, so that the liquid 2 to be examined is forced to flow through these capillaries and thereby the contact with the reagent or reagent mixture 20 can take place. These capillaries may further be designed to have a diameter which allows at least partial separation according to the molecular weight of the components of the liquid 2. This can be achieved because the DNA or RNA molecules usually have a higher molecular weight than the remaining substances in the blood that these molecules can not enter the interior of the capillaries and thus a separation of the DNA-20 or RNA molecules of the remaining components of the liquid to be analyzed 2 takes place and thus a fractionation by molecular weight is possible.

If the carrier 21 has a multilayer structure, in addition to the possibility that several individual components are analyzed in parallel, there is also the possibility that a layer for the lysis, a further layer for the stabilization and a third layer for the qualitative or quantitative separation of the used to analyze the substance.

Other layers for additional analysis steps are of course possible. The carrier 21 according to FIGS. 2 and 3 may for example have the form of a tablet.

FIG. 4 shows a variant embodiment of the collecting vessel 1, wherein the carrier 21 in the form of particles 25 is arranged in the vessel interior 3. These particles 25 may have permanent magnetic properties, e.g. be made of a Fe203, Ni core or the like. With plastic 35 or glass sheath, and be coated over at least part of its outer surface with the reagent or reagent mixture 20. The shape of the particles 25 can be chosen arbitrarily, for example spherical, lenticular, rod-shaped, etc. With suitable formation of the reagent or reagent mixture 20 on these particles 25, an affinity separation of the substances present in the liquid 2 can take place, ie Reagent mixture 20 can be selected so that there is a preferably particularly high affinity for the substance to be analyzed. In order, for example, to bind the DNA or RNA molecules to these supports 25, it is possible to provide a corresponding buffer in the interior of the vessel 3, with the aid of which the blood sample can be used, for example. be stabilized and the lysis of the cells can be done. 45

As already mentioned, it is possible, for example, for the reagent or reagent mixture 20 to cause the lysing of biological cells and thus, e.g. can cause stabilization of nucleic acids (DNA, RNA). In this case, the reagent or reagent mixture 20 may contain a guanidinium salt, e.g. a guanidinium halide, e.g. Guanidinium chloride, guanidinium thiocyanate, guanidinium carbonate, guanidinium nitrate, guanidinium acetate, guanidinium sulfate, guanidium stearate, guanidinium dihydrogen or hydrogen phosphate or the like, guanidinoalkyl sulfate esters, chaotropic reagents such as sodium iodide, potassium thiocyanate, ammonium sulfate or the like, urea, perchlorates or the like; Chloroform / phenol mixtures or the like. Included. In addition, it is of course possible, in order to maintain certain environmental conditions 55, that this reagent or reagent mixture 20 further buffer substances 1 2

AT 414 209 B and / or detergents, e.g. Tris (2,3-dibromopropyl) phosphate, tris (hydroxymethyl) aminomethane, ethoxylates of 4- (1,1,3,3-tetramethylbutyl) phenol, benzyltrimethylammonium hydroxide, (4- (2-hydroxyethyl) piperazino) ethanesulfonic acid, 3-morpholino-1-propanesulfonic acid, sodium dodecyl sulfate, polyoxyethylene derivatives of sorbitan esters, such as laurate, palmitate, 5-stearate, tristearate or oleate. Besides, reducing agents, e.g. Be contained ß-mercaptoethanol.

It is also possible that the vessel inner wall 5 is coated with reagents in the solid state, e.g. with anticoagulants such as EDTA, the latter also being presented as a solution.

Furthermore, the reagent or reagent mixture may contain citrate and / or phosphate buffer, as well as polydocanol as a detergent and dithiothreitol, TCEP as reducing agent. The guanidinium component may be present at a concentration between 0.05 M to 10 M and the buffer may be in the range between 1 mM and 500 mM. The detergent and the reducing agent may in turn be present in concentrations of 1 to 45 wt .-% and 0.1 to 15 wt .-%. The pH is preferably in the acidic range, for example between 3 and 6.5, but may also assume values of up to 8.

The advantage that can be achieved by the loosely presented particles 25 is a simple separation of the latter from the liquid matrix remaining in the collecting vessel 1. This separation may be e.g. done so that, as indicated by dashed lines in Fig. 4, a separation device 26 is inserted into the vessel interior 3. In order to avoid an unwanted spilling of the liquid 2, it is possible to provide this separation device 26 with the closure device 11, wherein the septum 12 can be dispensed with. In particular, it is possible to perform the closure device 11 as a cap in which the Separa-30 tion device 26 is supported.

If particles 25 having permanent-magnetic properties are selected as the carrier 21, it is possible, if the separating device 26 also has magnetic properties, to bind these particles to the separation device 26 on the basis of the laws of magnetism, which is shown in FIG the arrows 28 indicating a magnetic field 27 are shown schematically. The separation device 26 may in turn either have permanent magnetic properties or it is also possible to use the forces of electromagnetism. After the particles 25 have been bonded to the surface of the separation device 26, they can be removed from the liquid 2 and, for example, be transferred to a further collection vessel 1, where subsequently the particles 25 are released again from the separation device 26 and e.g. the analysis by detachment of the reagent or reagent mixture 20 with the substance from the carrier 21 and by subsequent detection of the substance allows 45 becomes light.

Of course, it is also possible that the reagent or reagent mixture 20 remains on the particles 25 and only the substance to be analyzed, e.g. DNA, RNA, is eluted. 50

Of course, it is also possible that this separation device 26 is carried out differently, for example, that this on the vessel bottom 10 side facing a device 29, as shown in Fig. 5, having, for example, the shape of a network. On this device 29, the particles 25 may be supported, so that on the one hand, the particles 25 55 when removing the separation device 26 from the collecting vessel 1 on the device 13th

AT 414 209 B 29 remain and on the other hand it is possible that excess liquid 2 remains through this network in the vessel interior 3 of the collecting vessel 1.

The particles 25 can be used as a loose bed of optionally multi-layered individual particles, e.g. Glass balls, silica gel particles or the like., To be arranged in the collection vessel 1. It is possible to arrange this loose bed in a predefinable height 24 from the bottom of the vessel 10, for which purpose a corresponding support device should be present in the collecting vessel 1. This support device may for example be designed as a perforated plate, which is supported by suitable means, for example by arranging the web 22, but also by the training io the collecting vessel of FIG. 3 on the predefinable height 24.

Furthermore, it is possible that the particles 25, e.g. are encapsulated in a separate shell, whereby a better cohesion of these particles 25 is possible and should this envelope be suitable for the passage of liquid, i. for example, have a corresponding perforation 15. It is also possible that this shell of absorbent materials, such as cellulose fibers or the like., Is. By this encapsulation of the particles 25, in turn, a simple removal of the latter can be made possible, so that in a subsequent operation, the analyte or reaction products of the reagent or reagent mixture 20 can be separated, for example by means of an Eluationsvorganges, 20 and is in turn the detection of to be analyzed substance without additional steps possible.

Figures 6 and 7 show possible embodiments for the carrier 21. This may e.g. a carrier body surface 31 may comprise, for example, the shape of a rod, a chain or the like, and it is possible to coat this carrier body surface 31 with the reagent or reagent mixture 20. Corresponding precautions can be taken to increase the surface area, for example by arranging surface enlargements 32 of at least approximately hemispherical design. Of course, these surface enlargements 32 can have any shape adapted to the particular application.

These surface enlargements 32 can either be connected immovably to the carrier body 30 or form part of the carrier body 30, for example by direct molding during the manufacturing process. 35

By means of these surface enlargements 32, it is furthermore possible to divide zonar the carrier body 30, whereby the arrangement of different reagent or reagent mixtures 20 on the carrier body 30 and thus a multi-component analysis becomes possible. Furthermore, it is possible for the particles 25 to be suitably fixed on the carrier body surface 31 and, in turn, for this purpose to be separated in a separate carrier particle receptacle, e.g. in already mentioned shell, be supported, whereby an easy removal of the particles 25 from the carrier body 30 is possible. The carrier particle reception is preferably designed for the passage of the liquid 2, whereby the reaction with the reagent or reagent mixture 20 becomes possible.

It is also possible that these particles have 25 capillaries in order, on the one hand, to increase the surface area of the particles 25 available for the reaction with the substance or, on the other hand, to effect a selective separation according to molecular weight, i. according to molecule size, to achieve so.

Of course, the carrier bodies 30 shown in FIGS. 6 and 7 can have any suitable, arbitrary shape. It proves to be advantageous if the carrier body 30 is elongated with two oppositely arranged

AT 414 209 B Neten carrier body ends 33 is formed. This makes it possible that at least one carrier body end 33, the closure device 11 is arranged, with any suitable connection between the carrier body 30 and the closure device 11 can be selected or it is of course possible that the closure device 11 with the support body 30 made in one piece is. It can thus be achieved that the carrier body 30 is inserted into the collecting vessel 1 so that an inadvertent spillage of the liquid 2 contained therein is prevented by sealing the vessel opening 4 of the collecting vessel 1. Thus, for example, intimate mixing of the liquid, e.g. by shaking, possible without the person operating the collecting vessel 1 running the risk, if appropriate with viruses and bacteria present in the liquid 2, e.g. HIV viruses, tuberculosis viruses, etc., to come in contact. For this purpose, the closure device 11 may for example be provided with a thread and e.g. be designed as a screw cap. Likewise, it is of course possible to exploit corresponding frictional forces between the closure device 11 and the surface of the collecting vessel 1 and thus form the closure device as a simple plug-in closure.

To improve the sealing of the vessel opening 4 by the closure device 11, it is of course possible that corresponding sealing elements 34 are arranged in the latter. These sealing elements 34 may be made of materials known from the prior art and may consist, for example, of elastomers. Furthermore, it is possible that the closure device 11 additionally comprises the septum 12. This allows the loaded carrier 21 to be removed from a first collection vessel 1 and placed in a second collection vessel 1 (not shown), and subsequent detachment of the substance to be assayed by insertion of a corresponding device, such as a cannula 25 (not shown) ), through the septum 12 a suitable solvent can be brought into the collecting vessel 1 and in turn there is the possibility, due to the collecting vessel 1 tightly closing closure device 11, the collecting vessel 1 for better and faster detachment of the loaded substance to be analyzed with the reagent or Reagent mixture 20 from carrier body 30 (or the species resulting from the reaction of the substance with the reagent or reagent mixture 20). In order to achieve a better distribution of a solvent to be supplied through the septum 12, the carrier body 30 may be provided with a channel 35 extending from the septum 12 at least approximately to the carrier body end 33 opposite thereto (both the septum 12 and the channel 35 are indicated by dashed lines in Fig. 6). 35

Furthermore, it is possible for this channel to branch off distribution channels which extend as far as the carrier body surface 31 or into the particles 12. It should be mentioned at this point that due to the affinity of RNA or DNA molecules to glass, it is of course possible to use carrier bodies 30 or carriers 21 which are not coated with the reagent 40 or reagent mixture 20.

It is also possible to use carrier bodies 30 which consist of a silicate material, cellulose, plastic or the like. Furthermore, it is possible to make the carrier body 30 from a gel, e.g. an agarose gel to manufacture.

Furthermore, it is possible that in the collecting vessel 1, in which the carrier body 30 is used, a corresponding reagent or reagent mixture 20 is presented, with the aid of such an appropriate preparation of the liquid 2 can take place, for example, the stabilization of the liquid 2, e.g. Blood, and / or the lysis of the cells causes.

However, the carrier 21 or carrier body 30 can also be made of an open-celled or closed-cell foam, which is impregnated with a corresponding reagent or reagent mixture 20 and which passes through the liquid 2 1 5 to be analyzed

AT 414 209 B allowed. This foam can be designed, for example, soft elastic and it is possible to manufacture this polyurethane. Of course, semi-hard to hard versions of the foam are possible. Fig. 8 shows, in part, a collecting vessel 1 in which the reagent or reagent mixture 20 is not initially charged as solid, but in the form of a corresponding liquid 36 or a liquid mixture and / or a solution of solids required for analysis in the corresponding solvents. The advantage that can be achieved thereby is that by feeding or introducing the liquid 2 to be analyzed into the collecting vessel 1, the reagent or reagent mixture 20 does not first have to be dissolved by the liquid 2 in order to be reacted be available with the substance to be analyzed. Since it is also possible to measure the prevailing in the receptacle 1 vacuum so that a pre-determinable amount of liquid 2 is sucked into the receptacle 1, the concentration of the reagent or reagent mixture 20 may always be present in excess, so that a rapid reaction of to be analyzed substance or substances with the reagent or reagent mixture 20 can take place and thereby optionally a stabilization of, for example, DNA or RNA molecules is not necessary. Of course, this information regarding the concentration may also apply in the case where the reagent or reagent mixture 20 is in the solid state. 20

Of course, it is necessary that in order to maintain the vacuum in the collecting vessel 1 over a longer period of time, as is necessary, for example, for storage of this collecting vessel 1, the closure device is designed accordingly. FIG. 9 again shows an embodiment variant of the collecting vessel 1 according to the invention, in which the carrier 21 is arranged in a collecting vessel extension 37, this collecting vessel extension 37 preferably having collecting vessel extension openings 38. One of these receptacle extension openings 38 is in communication with the vessel bottom 10 opposite Gefäßmantelstirnflächen 8 and can be connected, for example, a thread with the vessel shell 6 or, of course, other connection methods can be selected, for example, this connection can be designed as a plug connection. In any case, it should be ensured that this connection is so dense that the prevailing in the receptacle 1 vacuum is not degraded by incoming air. 35

The carrier 21 can now either be connected immovably to this collecting vessel extension 37 or it is possible to rest this carrier 21 on at least part of the vessel shell end faces 8 and to prevent only lateral slipping of the carrier 21 by means of the collecting vessel extension 37. 40

The second collection vessel extension opening 38 is preferably in turn closed with a closure device 11 - expedient manner with the septum 12 -. This makes it possible that by perforation of this septum 12, for example, with a cannula 19, not shown, the liquid 2 to be analyzed after its entry into the Sammelge-45 vessel 1 first hits the carrier 21, which in turn may contain the reagent or reagent mixture 20 , Run in the carrier 21, the required reactions and finally runs the unneeded liquid matrix in the lower part of the receptacle 1. For further analysis or for detachment of the reaction products or the substance to be analyzed from the carrier 21, this can either, if the carrier 21 is fixedly connected to the Sammelgefäßverso extension 37, placed together with this on a further collection vessel 1 and, for example, again by the Septum 12 of the closure device 11 are acted upon with a corre sponding solvent, so that the substance to be examined or the substance mixture to be examined runs into the further collection vessel 1 and is available there for the further process steps or the detection. 55 1 6

AT 414 209 B

However, if the carrier 21 is not immovably connected to the collection vessel extension 37, it is possible to transfer the carrier 21 separately into a further collection vessel 1 or to supply it directly to a final detection of the substances to be determined. In this way, it is possible to compose a complete analysis kit 39 for analyzing biological biological fluids or biological matrices, as shown in FIG. 10, for example. In this case, several collection vessel extensions 37 are arranged in the manner already described on the collecting vessel 1, these collecting vessel extensions 37 in turn, as already described, containing the carrier 21. A conclusion io forms the already described closure device 11, whereby on the one hand, the built-up in the receptacle 1 vacuum is obtained and on the other hand, the inlet of the liquid 2, not shown, is possible. In this way, by arranging a plurality of carriers 21 one above the other, a plurality of substances or mixtures of substances to be analyzed can be separated from the liquid 2 virtually simultaneously and thus these are available for further analysis.

However, this arrangement according to FIG. 10 can also be used so that the reagent or reagent mixture 20 (not shown) is separated into individual stages, so that the individual process steps required, such as stabilization and / or lysis of the cell and / or or the immobilization of the molecules to be studied, at different levels.

Of course, it is also possible in this case that the carrier 21, as already described, is designed to be multi-layered. 25

In Fig. 11, another embodiment for such an analysis kit 39 is shown. The collecting vessel 1 is in this case again from the vessel shell 6, but instead of the previously described vessel bottom 10, a further vessel opening 4 is present. However, it is also possible, as shown in dashed lines in the lower part of FIG. 11, that in the vessel shell 6 a further vessel opening 4, e.g. with the cannula 19, is arranged.

The two vessel openings 4 are preferably closed by the closure device 11, which contains the septum 12. As a result, on the one hand it can be achieved that the liquid 2 to be analyzed 2 is supplied to the vessel interior 3 via a corresponding device, for example the cannula 19. On the other hand, it is also possible thereby to connect a plurality of these collecting vessels 1 via a cannula 19, in turn, as shown in FIG. 11. Such a design makes it possible to arrange different reagents or reagent mixtures 20, not shown, in the various collection vessels 1, wherein this can optionally be done in solid or liquid form with the aid of which a separation into components or fission products of the liquid 2 can be done. After separation of the individual collection vessels 1, these are thus automatically available separately for a further analysis without additional manipulation steps.

Of course, it is also possible in this variant of the analysis kit 39 to arrange in the collecting vessels 1 carrier 21, which can be removed through the respective vessel opening 4, but are not shown in Fig. 11. 50

It is also possible with this method to equip the analysis kit 39 in such a way that one of the collection vessels 1 is presented with the eluent required for the elution of the substance to be analyzed by the carrier 21 and this after the last mentioned collection vessel 1 has been assembled with one or more carriers 21 containing collecting vessels 1 runs over the 55 respective carrier 21 and thus causes the eluation. 17

AT 414 209 B

It is further possible for the analysis kit 39 to be designed such that several of these collection vessels contain salt solutions, e.g. Cesium chloride solutions, different density, whereby a separation of the liquid to be analyzed 2 by centrifugation, preferably at high g-values, using a cesium chloride gradient is possible. It goes without saying that the corresponding collecting vessel 1 is connected to the desired cesium chloride concentration after each centrifugation step with the collecting vessel of the previous cesium chloride concentration.

In Fig. 12, in turn, a variant of the collecting vessel 1 is shown, which can be provided with 2 Gefäßöffnun-gene gene 4, which are closed with the septum 12 containing closure device 11. In the interior of the vessel 3, a device 40 is arranged, by means of which a physical separation of the constituents of the liquid 2 during or after the centrifugation can be effected. This device 40 is dimensioned in its density so that the density of one of the two phases formed during centrifugation is greater and the density of the other phase is lower, so that this device 40 comes to rest at the interface of the two phases.

In order to allow phase separation, this device 40 may have a schematically indicated, after centrifugation again occlusive device opening 41, 20 by which it is the phase with the lower specific weight allows to rise in the upper part of the receptacle 1.

However, the device 40 can also be designed so that it does not abut the vessel shell 6 over the entire circumference during centrifugation, i. on the vessel inner wall 5, so that 25 thus the phase with the lower specific weight between the vessel inner wall 5 and the device 40 can reach into the upper part of the collecting vessel 1.

It goes without saying that the tube-like connections shown in FIG. 12, for example the cannulas 19, via which a plurality of collection vessels 1 can be connected, are not attached to the collecting vessel 1 or the septum 12 during the centrifugation.

By separating the two phases and by the accessibility of the collecting vessel 1 via at least two vessel openings 4, the two phases can be separated from each other in a simple manner, so that an after treatment of at least one of the phases with a corresponding reagent or reagent mixture 20 in the same or a connected to this receptacle 1 further collecting vessel 1 is possible.

Through the two vessel openings 4 but also access to a arranged in the collection vessel 1, in Fig. 12 but not shown carrier 21 made from both sides. 40

If the phase separation leads to more than two different phases, then it is possible to arrange at least one further vessel opening 4 along the vessel shell 6, as already described, which in turn may be closed by a corresponding closure device 11. In the case where a plurality of collection vessels 1 are connected to an analysis kit 39 as shown in FIG. 10, there is a possibility that the vacuum in the collection vessel 1 into which the liquid 2 to be analyzed last enters is selected so that the liquid 2 is sucked through all of the collecting vessels 1 present. 50

FIG. 13 shows an embodiment variant of the collecting vessel 1, in which a conventional collection vessel, for example a blood collection tube, is connected to a collecting vessel 1 according to the invention. The connection can in turn via a thread, a plug-in closure or the like. Take place or it is also possible that between the two collection 55 vessels several receptacle extensions 37 (not shown in Fig. 13) arranged 18th

AT 414 209 B are.

This design makes it possible to first collect the liquid 2 to be analyzed in the conventional collecting vessel 1 and to stabilize it there for any subsequent analysis and to store it therewith.

In the collecting vessel 1 according to the invention, in turn, the carrier 21 can be arranged and it is possible that by a tilting movement of this arrangement, the liquid 2 passes through the carrier and thus the substance to be analyzed adheres to the carrier and is available for further analysis io available.

Of course, the reverse process is possible, namely, that arranged on the support 21 reagent or reagent mixture 20 selectively binds or reacts with those substances which are not to be analyzed, so that only the sub-15 to be analyzed mixture or die analyzing substance passes through the carrier 21.

Fig. 14 shows an arrangement of the carrier 21 in the course of the already described tubular connection, for example the cannula 19, by means of which e.g. the blood of a patient can be removed. This carrier 21 may in turn be loaded with the reagent or reagent mixture 20.

It is possible in this way, since the carrier is now not connected to the collecting vessel 1 or contained in the vessel interior 3, the substance to be analyzed simply from the liquid matrix, which runs after passing through the carrier 21 in the collecting vessel 1, 25 with which is connected to the cannula 19 via the septum 12, or it is therefore not necessary to open the receptacle 1 to remove the carrier 21 from the vessel interior 3 and thus again a higher level of security for the operating personnel can be achieved. FIG. 15 shows an embodiment variant of the collecting vessel 1 in which the carrier 21 is connected directly to the septum 12 of the closure device 11, for example, being integrally formed thereon. Of course, other connection methods are possible.

In order to enable the reaction of the substances contained in the liquid 2 with the reagent or reagent mixture 20 contained on the carrier 21, the tubular connection, for example the cannula 19, should not be completely pushed by the carrier 21.

With this embodiment, it is possible with the removal of the closure device 40 11 from the collection vessel 1 at the same time to remove the carrier 21 with the substances to be analyzed.

Finally, FIG. 16 shows a variant embodiment in which an inner collecting vessel 1 is contained in an outer collecting vessel 1 or can be inserted into the outer collecting vessel 1.

The inner collecting vessel 1 has at least one vessel jacket opening 42 in the vessel jacket 6. The connection of the interior with the outer receptacle 1, for example, by a sliding closure 43, which is arranged at the vessel opening 4 of the outer receptacle 1 or screwed to the vessel shell 6 of the outer receptacle 1, take place and this slide closure 43, for example, as from the Prior art known Quickfit be executed. Through the central opening, which has this sliding closure 43, the inner receptacle 1 can be inserted into the outer receptacle 1. 55 19

AT 414 209 B

If a liquid 2 to be analyzed is placed in the outer collecting vessel 1, the insertion of the inner collecting vessel 1 into the outer collecting vessel 1 produces a corresponding pressure on the liquid 2, so that this, if a diameter 44 is chosen to be smaller than a collecting vessel inner diameter 45 of the outer collecting vessel 1, of course, the wall thickness of the inner receptacle 1 must be taken into account, escapes into the gap between these two receptacles 1 and subsequently enters through the vessel shell openings 42 in the inner receptacle 1 in the vessel interior 3 of the inner receptacle 1. If the inner collecting vessel 1 is coated on its outside with a reagent or reagent mixture 20, the liquid 2 or its components can react with this reagent or reagent mixture 20 on the way from the outer collecting vessel 1 into the inner collecting vessel 1, so that individual Components or cleavage products or mixtures of substances can be deposited thereon.

Now, if a sliding closure seal 46 is designed so that upon withdrawal of the inner from the outer receptacle 1, the reagent or reagent mixture 20 is stripped with the substance thereon or the mixture of substances from the outer vessel surface of the inner receptacle 1, so it can be an automatic Separation of the substances to be analyzed done. Of course, should the reagent or reagent mixture 20 interfere with the analysis, in particular when it is selected as a carrier 21, be separated from the substances to be analyzed in the sequence.

This embodiment variant of the collecting vessel 1 or analysis kits 39 is particularly suitable even if the liquid to be analyzed is temporarily stored for the time being and the analysis takes place only at a later point in time.

The reagent or reagent mixture 20 may be selected or composed so that the lysis and / or stabilization of individual components of the liquid 2 or the biological matrix is effected. Likewise, it is of course possible that this reagent or reagent mixture 20 or a part of this Regenzgemisches 20 is an extractant, e.g. a phenolic solution, a phenol / chloroform mixture or the like, in order to separate proteins from the DNA, for example, by means of which, as usual with extractants, a part of the liquid 2 or the biological matrix is transferred into this extractant. According to Neernst's law, the extractive agent should be chosen so that, if possible, by a one-step " shake-out " the concentration of analyte in the residual fluid matrix approaches zero.

For stabilization, especially when the liquid 2 is blood, the reagent or reagent mixture 20 may contain a guanidinium salt.

Furthermore, the reagent or reagent mixture 20 may at least partially contain cesium chloride in order to be able to carry out a cesium chloride centrifugation known from the prior art.

However, the reagent or reagent mixture 20 can also be chosen so that the liquid 2 or components of the biological matrix are divided into several phases.

It is further possible that the reagent or reagent mixture 20 causes a selective precipitation of individual constituents of the liquid 2 or the biological matrix, in order to be used e.g. to allow a purification of the nucleic acids. Thus, the reagent or reagent mixture 20 may be selected so that the DNA or RNA molecules or cleavage products thereof are deposited. On the other hand, it is also possible that, for example, by the so-called salting-out effect or changes in the pH, a selective precipitation of proteins takes place.

Thus, it is possible, for example, that one of the collection vessels according to the invention 1 Isopro-

AT 414 209 B panol or ethanol contains, in addition to the precipitation in addition to achieve a concentration of DNA or RNA molecules or fission products thereof or individual other components of the liquid 2 or the biological matrix. Should it be expected that the concentration of nucleic acid is low, it is also possible to mix reagent or reagent mixture 20 with an inert carrier 21, for example glycogen, in order to increase the efficiency of precipitation formation.

However, the reagent or reagent mixture 20 can also consist of lithium chloride or contain lithium chloride. Lithium chloride is e.g. preferably used when only RNA is to be knocked down and not proteins or DNA.

In the case of permanent magnetic supports 21, it is possible that they are coated with, for example, streptavidin, e.g. for the analysis of poly (A) + mRNA. Other compositions of the reagent or reagent mixture 20 are possible in order to specifically separate or adsorptively bind other nucleic acids as well. However, it is also possible that the reagent or reagent mixture 20 is a gel, e.g. an agarose gel, a plastic gel, or the like, is such that gel filtration may subsequently be performed, the gel being a matrix of defined pore size. Such molecules, whose diameter is smaller than the defined pore size, can thus diffuse into these pores, whereas size-20 ren molecules is denied entry and they can thus be separated. The molecules which have penetrated into the pores may subsequently be delayed in time according to their molecular size, e.g. eluted according to increasing molecular size.

In addition to the above-mentioned selection for the reagent or reagent mixture 20, the latter can of course be chosen so that a purification of the nucleic acids can take place, for example by chromatography, electrophoresis, centrifugation, affinity deposition or the like. possible that, as already mentioned, a carrier 21 made of glass is used, since the DNA molecules have an affinity for glass and so e.g. can be separated from proteins and RNA. 30

Furthermore, it is possible to form the reagent or reagent mixture 20 from endo- and / or exonucleases in order to enable a structural clarification of the DNA or RNA. The endonucleases may be, for example, deoxy- and / or ribonucleases, where both enzyme groups cleave DNA or RNA, but only if they consist of 3 ', 5' -phosphorus-diester compounds. For example, it is possible to convert high molecular weight DNA into nucleotides with the two DNAses I or II, whereby DNAse I leads to tri- and tetradesoxin-nucleotides having a 5-substituted phosphate group. DNAase II forms oligodeoxynucleotides with 3 'phosphate ends. Of course, it is also possible that the reagent or reagent mixture 20 forms a corresponding buffer in order to provide a suitable medium for analysis or deposition, stabilization, immobilization, etc. available.

Furthermore, it is possible with the aid of the reagent or reagent mixture 20 to carry out an indirect detection, for example by the biotin / streptavidin amplification method, wherein this reagent or reagent mixture should contain at least 2 antibodies.

The use of the collecting vessel 1 or of the carrier 21 or of the analysis kit 39 can now be carried out so that the reagent or reagent mixture 20 already during the introduction of the liquid 2 into the collecting vessel 1 or the biological matrix or at least individual constituents of which are separated and / or immobilized and / or stabilized. For further separation of the liquid 2, a plurality of collecting vessels 1 can be connected to one another, as has already been explained. If the reagent or reagent mixture 20 does not already cause a separation of individual components from the liquid 2 or the biological matrix 55, it is possible with the aid of the carrier body 30, this separation by the 21st

AT 414 209 B subsequent introduction into the collecting vessel.

FIG. 17 shows a further embodiment of the collecting vessel 1 according to the invention. Although designed in a single-layered manner, this possibility, as well as all variants described above, makes it possible to design the collecting vessel 1 in one or more layers, ie. So that the vessel shell 6 can be designed as a laminate of different plastics. It is also possible to select the plastics in such a way that at least one assumes the supporting function of the collecting vessel 1 and another plastic is carried out correspondingly tight with respect to the permeation of liquids and / or gases. For example, it is possible for the part of such a laminate of polypropylene, which faces the vessel interior 3, and the outer layer of this laminate, which is preferably formed thereon, to consist of polyethylene terephthalate.

There is also the possibility of carrying out the vessel jacket 6 of the collecting vessel 1 not as a laminate 15 of different plastics, but in the form of at least two individual vessels pushed one inside the other, which in turn are made of different plastics, e.g. above requirements can exist.

In the embodiment according to FIG. 17, it is shown how the particles 20, which in turn can be embodied, for example, as permanent-magnetic particles with or without coating, together with the substance to be analyzed from the matrix, e.g. the removed blood, can be removed. For this purpose, the closure device 11, for example a screw cap, a finger-like, preferably made of plastic, glass or the like. Forsatz existing 47 have. Preferably, this extension 25 47 is made of transparent material and has an opening 48 towards the environment.

The along the vessel central axis 7 of this opening 48 opposite, preferably the vessel bottom 10 adjacent side of the extension 47 is preferably closed.

With the help of this extension 47, it is possible, for example, a rod 49 inserted therein ren, which may have magnetic properties. Under the influence of the magnetic field emanating from this magnetic rod 49, the particles 25 are now bound to an outer surface 50 of the extension 47 and thereby removal of these particles 25 together with the substance to be analyzed from the liquid 2, for example the closure device 11 is removed. 35

In a further embodiment, it is possible that a holding device 51 is arranged on the closure device 11, by means of which the rod 49 can be arranged at least approximately immovable in the extension 47. This holding device 51 may be embodied, for example, in the manner of a seal, wherein over the frictional forces between the Halteeinrich-40 device, for example, arranged in this elastomer, and the rod 49, a corresponding support can be achieved. The holding device 51 is indicated by dashed lines in Fig. 17 and, for example, the closure device 11 may be formed in this case again as a Quickfit. Of course, it is also possible in this embodiment of the receptacle 1, that the closure device 11, the septum 12 (not shown in FIG. 17), so that a subsequent replacement of the closure device by those of the type described for Fig. 17 is omitted and can, for example the septum 12 may be formed annularly around the extension 47. In the event that this collection vessel 1 according to FIG. 17 is used for blood collection with simultaneous lysis of the cells and stabilization of the nucleic acids in order to prevent degradation of the latter, a solution, as already described, may preferably be present in the collecting vessel 1 a chaotropic substance, eg a guanidinium salt, a buffer, a detergent, and preferably a reducing agent. In this case 22

AT 414 209 B, it may be sufficient if the particles 25 are designed only as permanent magnets and a coating of the latter is dispensed with, if a selective analysis for certain nucleic acids is not required. For this purpose, as already mentioned, these particles 25 may be multi-layered and comprise, for example, a magnetic core 5 of Fe 2 O 3 or nickel or the like, which is surrounded by another layer, for example of plastic or glass. On the other hand, it is of course possible that the layer structure is chosen differently, namely from a carrier substance for the core, which is coated with a magnetic material and in turn a polymeric layer is arranged. 10

Instead of a closure device 11, as shown in Fig. 17, with the extension 47, it is possible to use a conventional, provided with or without septum 12 closure device 11 and the separation of the preferably magnetic particles 25 by means of a from the outside to the collecting vessel. 1 make the magnet mentioned. In this case, the separation of the liquid residual substance takes place by removing the closure device 11 and emptying the collecting vessel 1.

It is also possible, the collection vessel 1, in particular the closure device 11, to be designed such that the extension 47 is not formed on the Verschußvorrichtung 11 but 20 rather can be removed. This makes it possible for this extension 47, together with the magnets and the particles 25 adhering to the extension 47, to be automatically, e.g. in an automatic system for analysis, and for example to transfer the particles 25 with the substance to be analyzed thereon into another reaction vessel. The detachment of the particles 25 from the extension 47 can take place, for example, by pulling the rod 49 out of the extension 47. In order to prevent in this embodiment, a stripping of the particles 25 upon removal of the extension 47 from the closure device 11, it is possible to remove a portion 52 of the closure device 11 to the extension 47 together with the latter. This subregion 52 is indicated in phantom in FIG. 17, and if this is possible, e.g. to provide a predetermined breaking point in the closure device 11 30.

In addition to the materials already mentioned for these particles 25, the latter can also consist of porous glass, for example of silicon dioxide. This has the advantage that, as a result of the chemical inertness of this material, organic as well as aqueous solvents can be used over a wide pH range even in the presence of aggressive reagents or reagent mixtures. Furthermore, in both these and all other described particles 25, it is possible to coat them. For example, in the case of the glass particles, the surface may be provided with hydrophilic and / or hydrophobic reactive groups, e.g. be bound to the porous glass via covalent siloxane bonds.

Furthermore, it is possible to produce these particles 25 of polystyrene.

The diameter of the particles 25 may be in the range between 0.1 pm and 250 pm, for example 45 between 0.4 and 100 pm, in particular between 1.0 pm and 50 pm. For individual applications, however, the largest grain may also have a diameter of more than 500 μm.

In a further embodiment it is possible to use the closure device 11 with a permanent magnet or e.g. to equip an electromagnet. This makes it possible, so to bind the permanent magnetic particles 25 by simply turning the collection vessel 1 to the closure device 11, and these can be safely removed together with the closure device 11 from the collection vessel. Particularly advantageous is the use of an electromagnet, for example, by installing a coil in the closure device 11 with corresponding connections for power supply, since thus an unintentional adhesion of the particles 25 on the closure device 11 before their actual 23

AT 414 209 B use can be prevented. By appropriate design of the closure device 11, a subsequent cleaning of these and optionally a sterilization is possible and thus the closure device 11 can be used several times. In FIGS. 18 and 19, a further embodiment of the collecting vessel 1 according to the invention with a closure device 11 is shown schematically simplified. The closure device 11 is used in this embodiment for holding at least two analysis rods 53, which may optionally be coated with a variety of reagents or reagent mixtures 20. It is thereby a multi-component analysis possible. 10

These analysis rods 53 may either be connected immovably to the closure device 11 so as to be removed from the liquid 2 together with the closure device 11 and the substances to be analyzed. On the other hand, it is of course possible that these analysis rods 53, as indicated by dashed lines in Fig. 18, again 15 are supported by holding means 51.

As FIG. 19 shows better, these analysis rods can be arranged symmetrically around the preferably present septum 12 in the closure device 11, so that the separation of various substances to be analyzed already during filling of the collecting vessel 1 20 through this septum 12, in particular through a cannula 19 (in Fig. 19 not shown) begins.

These analysis rods 53 are preferably designed as carriers for reagents or reagent mixtures 20 already described, and this embodiment variant of the collection container 1 according to the invention can be used, for example, for the qualitative analysis of e.g. Blood be used. It is possible that the reagents or reagent mixtures 20 different, preferably color indicators include, which can be concluded solely from the successful or failed color change on the presence or absence of the sought substance. 30

It is also possible that in the event that the closure device 11 is designed as a magnetic carrier, for example, pyroelectric or piezocrystals are arranged in this, so that in the sequence required for a corresponding solenoid current is generated by increasing the temperature or by pressurization. 35

For the sake of the order, it should be pointed out that, for a better understanding of the structure of the collecting vessel, the carrier and the analysis kit, these or their components have been shown partly unevenly and / or enlarged and / or reduced in size. The problem underlying the independent inventive solutions can be seen from the description.

Above all, the individual in Figs. 1; 2; 3; 4; 5; 6; 7; 8th; 9; 10; 11; 12; 13; 14; 15; 16; 17; 18, 19 shown embodiments form the subject of independent, inventive solutions 45. The relevant objects and solutions according to the invention can be found in the detailed descriptions of these figures.

Reference designation so 1 collection vessel 2 liquid 3 vessel interior 4 vessel opening 5 vessel interior wall 41 device opening 42 vessel shell opening 43 sliding closure 44 diameter 45 collection vessel diameter 55

Claims (41)

24 AT 414 209 B 6 Vessel sheath 7 Vessel axis 8 Vessel sheath end face 9 Vessel sheath end face 10 10 Vessel bottom 11 Closure device 12 Septum 13 Outer jacket 10 14 Bore 15 Outer shell inner side 16 Thread 17 Bar 15 18 Vessel shell outer wall 19 Cannula 20 Reagent mixture 21 Carrier 20 22 Bar 23 Dimensional change 24 Height 25 Particles 25 26 Separation device 27 Magnetic field 28 Arrow 29 Device 30 30 Carrier body 31 Carrier body surface 32 Surface enlargement 33 Carrier body end 34 Sealing element 35 35 Channel 46 Slide seal 47 Extension 48 Opening 49 Rod 50 Surface 51 Holding device 52 Subsection 53 Analysis bar 36 Liquid 37 Sump extension 38 Sump extension 40 39 Analysis Kit 40 Apparatus Claims: 45 1. Collection container for liquids for use in nucleic acid analysis, eg Blood collection tube, with one of a vessel shell and optionally a vessel bottom, which (r) a vessel inner wall (t), at least partially surrounded vessel interior space by at least one vessel opening which is closable with a septum verse-50 hene closure device is accessible , characterized in that a lysis reagent is introduced into the interior of the vessel (3) and at least one reagent or reagent mixture (20) with which components of the liquid (2) disrupted by the lysis are separated off or immobilized and / or stabilized, on a support (21 ) is arranged. 55 25 AT 414 209 B 2. Collection vessel according to claim 1, characterized in that the reagent or reagent mixture (20) selectively for nucleic acids, nucleic acid mixtures or nucleic acid components, e.g. Mononucleotides, oligonucleotides, is. 3. A collecting vessel according to claim 1, characterized in that the reagent or reagent mixture (20) selectively for nucleic acid degrading components of the liquid (2) or the biological matrix, e.g. Proteins, metabolites, nucleases, enzymes or the like. 4. Collection vessel according to one of the preceding claims, characterized in that io the reagent or reagent mixture (20) is present in the solid state. 5. Collection vessel according to one of the preceding claims, characterized in that the reagent or reagent mixture (20) is present as a liquid or liquid mixture and / or solution. 15 6. Collection vessel according to one of the preceding claims, characterized in that a part of the reagent mixture (20) is an extractant, e.g. a phenolic solution, a phenol / chloroform mixture. A collecting vessel according to any one of the preceding claims, characterized in that the reagent or reagent mixture (20) contains at least one guanidinium salt, e.g. Guanidinium chloride, guanidinium thiocyanate, guanidinium carbonate, guanidinium nitrate, guanidinium acetate, guanidinium sulfate, guanidinium stearate, guanidinium dihydrogen phosphate, guanidinium hydrogen phosphate, guanidinoalkyl sulfate ester. 25 8. Collection vessel according to one of the preceding claims, characterized in that the reagent or reagent mixture (20) contains cesium chloride. 9. Collection vessel according to one of the preceding claims, characterized in that the reagent or reagent mixture (20) contains ethanol and / or lithium chloride. 10. Container according to one of the preceding claims, characterized in that the reagent or reagent mixture (20) is a gel, e.g. an agarose gel, a plastic gel, is. 35 11. Collection vessel according to one of the preceding claims, characterized in that the carrier (21) is designed in the form of an inert filter. A collecting vessel according to claim 11, characterized in that the filter is separated by a loose bed of single particles, e.g. Glass beads, silica gel particles or the like. Is formed. 13. Collection vessel according to claim 11 or 12, characterized in that the filter has capillaries. 14. Collection vessel according to claim 13, characterized in that the capillaries have a diameter which allows at least partial separation according to the molecular weight of the components of the liquids (2) of biological origin. A collecting vessel according to any one of claims 1 to 10, characterized in that the carrier (21) is made of glass, e.g. Glass fibers, silicate materials, cellulose, latex, plastic, foam. Collection vessel according to one of the preceding claims, characterized in that the carrier (21) is in the form of particles (25) with e.g. permanent magnetic properties 55 is formed. 26 AT 414 209 B 17. Collection vessel according to one of the preceding claims, characterized in that the carrier (21) or the reagent or reagent mixture via two opposing closure devices (11) is accessible. 18. Collection vessel according to one of the preceding claims, characterized in that the carrier (21) or the reagent or reagent mixture (20) via at least one further vessel opening in the vessel shell along the vessel central axis is accessible. 19. Collecting vessel according to one of the preceding claims, characterized in that in the vessel interior (3) a device (40) is arranged, which physical separation of the components of the liquid (2) of biological origin or the biological matrix, for. after centrifugation causes. 20. Collection vessel according to one of the preceding claims, characterized in that the vessel shell 15 and / or the vessel bottom is formed by a plastic with polar or polari sierbaren groups. 21. A carrier for reagents or reagent mixtures for use in a collecting vessel according to one of claims 1 to 20, with a carrier body with a Trägerkörperoberflä- 20 che, characterized in that on the carrier body (30) a reagent or reagent mixture (20) is arranged, with which at least individual components of a liquid (2) of biological origin are separated or immobilized and / or stabilized. 22. A carrier according to claim 21, characterized in that the carrier body surface (31) 25 with the reagent or reagent mixture (20) is coated. 23. A carrier according to claim 21 or 22, characterized in that the carrier body consists of individual particles (25). 24. A carrier according to claim 23, characterized in that the particles (25) have capillaries. 25. A carrier according to any one of claims 21 to 24, characterized in that the carrier body (30) is formed in the form of chain links. 35 26. A carrier according to any one of claims 23 to 25, characterized in that the carrier body (30) comprises a holding device, e.g. a rod, to which the particles (25) are attached. 27. A carrier according to any one of claims 23 to 26, characterized in that the particles (25) are designed as permanent magnets. 28. A carrier according to any one of claims 23 to 27, characterized in that the particles (25) are supported by a carrier body particle receptacle. 45 29. A carrier according to any one of claim 28, characterized in that the carrier body particle receiving is formed lattice-shaped. 30. A carrier according to claim 21 to 22, characterized in that the carrier body 30 is longitudinally 50 Lieh with two mutually opposite carrier body ends (33) is formed, and at one of the carrier body ends (33) a closure device (11) is arranged. 31. A carrier according to claim 30, characterized in that the closure device (11), e.g. designed as a screw cap or plug closure. 55 2 7 AT 414 209 B 32. A carrier according to claim 30 or 31, characterized in that the closure device (11) comprises a septum. 33. A carrier according to claim 32, characterized in that the carrier body (30) with a 5 channel (35) is provided which extends from the septum (12) to at least approximately to the sem opposite carrier body end (33). 34. A carrier according to claim 33, characterized in that branch off from the channel (35) distribution channels, which extend to the carrier body surface (31). 10 35. A carrier according to claim 21, characterized in that the carrier body (30) is a gel. 36. A carrier according to any one of claims 21 to 34, characterized in that the carrier body (30) consists of glass, a silicate material, cellulose or a plastic, foam material. 37. Analysis kit for analyzing liquids of biological origin with at least one collecting vessel for holding the liquid or constituents of the liquid and at least one carrier for holding a reagent or reagent mixture, characterized in that the collecting vessel (1) according to one of claims 1 to 20 and / or the carrier (21) according to one of claims 21 to 36 is / are formed. Analysis kit according to claim 37, characterized in that at least one further collecting vessel is connected via a tubular connection, e.g. a cannula (19) connected to the collection vessel 25 (1). 39. Analysis kit according to claim 37 or 38, characterized in that for connecting the tubular connection to the collecting vessel (1), a septum (12) is arranged. 40. Analysis kit according to one of claims 37 to 39, characterized in that a Sam melgefäß (1) contains an eluent for the elution of the carrier (21) bound components of the liquid (2) of biological origin. 41. Analysis kit according to one of Claims 37 to 40, characterized in that the seminal vials (1) contain a salt solution, e.g. a cesium chloride solution, wherein the density of these solutions in the individual collection vessels (1) is different. For this purpose 5 sheets drawings 40 45 50 55