Title of Invention

DEVICE AND METHOD FOR SIMULTANEOUSLY IDENTIFYING BLOOD GROUP ANTIGENS

Abstract Device for the simultaneous and qualitative or quantitative determination of a plurality of analytes in a liquid sample, comprising a membrane 2 with 5 - an application zone 5 for the application of the liquid sample, at least one group of at least two indicator zones, which are able to interact with the analyte(s) and at least one absorption region 3 which takes up the liquid after having passed the indicator zones 10 wherein the indicator zones are located between the application zone 5 and the absorption region 3, characterized in that the flow directions from the application zone 5 through the respective indicator zones of a group towards an absorption region 3 (flow tracks) are substantially parallel and that at least two different flow tracks are present. 15 Furthermore, the invention relates to a method for the determination of a plurality of analytes or their derivatives in a liquid sample, comprising: the application of the sample onto the application zone 5 of a membrane 2 of the device according to any one of the preceding claims 1 to 8, wherein this sample is 20 present in adequate amounts in order to induce the test liquid to flow in the direction of the absorption region 3 through the indicator zones and to induce the analytes or their derivatives in the test liquid to form a complex in the indicator zones.
Full Text Device and method for simultaneously identifying
blood group antigens
The invention relates to a device for lateral-diagonal flow multi-parameter tests, in
particular in the field of blood group serology, for the simultaneous, qualitative or
quantitative determination of a plurality of analytes in a liquid sample, including a
membrane with an application zone for the application of the liquid sample, at least two
indicator zones which are able to interact with the analyte(s) and at least one absorption
region which absorbs the liquid after having passed the indicator zones, the indicator
zones being positioned between the application zone and an absorption region,
characterized in that the directions of flow from the application zone through the
respective indicator zones to an absorption region (flow tracks) are essentially parallel
and there being present at least two different flow tracks.
The invention further relates to a process for analyzing a plurality of analytes in a liquid
sample, comprising applying the sample onto the application zone of a membrane of the
apparatus according to the invention, wherein this sample is present in abequate quantity
in order to induce the sample liquid to flow in the direction of the adsorption region
through the indicator zones and in order to induce the analytes or their derivatives in the
sample liquid to form a complex in the indicator zones, in particular for the simultaneous
determination of cellular and plasmatic parameters, preferably for the simultaneous
conduction of blood group antigens.
In blood group serological diagnostics, parameters are generally tested which are of
particular relevance in the context of transfusions or of morbus haemolyticus neonatorum
(Mhn). This includes inter alia the detection of antigens on the surface of erythrocytes
which are characteristic for the blood groups. Further important antigen systems are
present also on thrombocytes, granulocytes, lymphocytes which likewise play a role in the
context of transfusions and/or transplantations.
It is known that for determining blood group antigens the erythrocytes of the persons to
be tested, (donors or recipients) are brought together with reagents which contain blood
group specific antibodies. Generally these tests are performed in the liquid state, in which
by mixing of an eirthrocyte-containing sample with a sample containing antibodies
directed against a specific blood group characteristic a testing batch is produced. The
testing batch is then incubated over a defined period and under defined conditions and
Oafter conclusion of the incubation, either directly or after a centrifugation step, is tested
visually or by optical methods for a possible agglutination or adsorption of the
erythrocytes. The predominant end point measurement in blood group serology is still the
haemaglutination test. For each blood group to be determined a separate batch must be
pipetted, i.e. e.g. for the determination of the nine most important blood groups A, B, D,
C, c, E, e, Cw and K, nine separate batches are needed, without counting any control.
Lateral flow tests nowadays are frequently applied as quick tests e.g. as pregnancy tests,
for determining infection markers or for drug screening. A lateral flow test device in a
known manner includes a rigid support on which an application zone for the sample to be
tested is provided, a separating membrane, on which bonding elements, e.g. catcher
antibodies or antigens are bound and on which the bonding reactions can be detected, and
a suction generating absorption region which causes the sample to be tested to flow in a
linear manner through the separating membrane.
Test membranes of conventional lateral flow tests are generally described involving a
chromatography-like separation. The analyte in the sample bonds specifically to the
bonding elements fixed in a membrane which as a rule are present in consecutive or
superimposed bands serving as indicator zones. The bonding complex is rendered visible
by indicator particles which as a rule are already present in the device in dehydrated form
in a conjugate liberation pad. The conjugate liberation pad is typically provided between
the application zone and the membrane. The pre-coated coloured indicator particles are
coated for example with an antibody directed against the analyte to be tested for.
The conventional lateral flow test format corresponds to a so-called "sandwich assay", in
which both the indicator zone as well as the indicator particles are coated with a ligand
aimed at the analyte tested for, normally an antibody. In that context the ligand (bonding
element) is immobilized on the membrane. The detector reagent, normally an antibody
bonded to a coloured polystyrene particle or to colloidal metals, is deposited in the
conjugate liberation pad in a leachable manner. This bonding complex serves as indicator
particle. Once the sample to be tested has been applied it very rapidly wets the conjugate
liberation pad, whereby the indicator particles are mobilized. The indicator particles
migrate with the liquid front along the porous membrane. An analyte present in the
sample becomes bonded by the antibody coupled to the indicator particle. As the sample
passes the indicator zone, the analyte/indicator particle complex in the indicator zone is
immobilized by reaction of the analyte with the antibody bonded in the indicator zone,
resulting in a visible signal.
A further known test format for small analytes comprising but a single antigenic
determinant, incapable of simultaneously bonding two antibodies, is the so-called
20"competition assay". The detector reagent bonded to the indicator particle is normally a
molecule identical to or analogous with the analyte. The indicator particles are deposited
in the conjugate liberation pad. The indicator particles migrate with the liquid front along
the porous membrane. If the sample contains the analytes, and if the indicator particles
(which effectively likewise contain analyte) pass the indicator zone, part of the analyte
molecules in the sample bond to part of the indicator particles. The more analyte is
present in the sample the more effective will it compete with the bonding of the indicator
particle and the weaker will the signal become.
According to the prior art these indicator particles are predominantly composed of
colloidal gold or of polystyrene, manufactured and coated according to methods known to
the skilled person. In the typical lateral flow test formats the analytes are determined
indirectly. In this context a direct determination of an analyte denotes that the analyte is
already bonded naturally to the indicator particle (e.g. erythrocyte). In the more common
situation of indirect determination of the analytes the sample to be tested as a rule
contains a non-cellularly bonded, e.g. plasmatic component as the analyte and, besides the
sample to be tested, two reagent components are required, i.e. indicator particles and a
bonding element. In the indirect determination the analyte initially bonds to the indicator
particle dissolved out of the conjugate liberation pad, before this complex becomes
immobilized in the indicator zones with the bonding element by way of a second reaction.
When using conventional lateral flow tests with erythrocytes as indicator particles which
have been bonded to the analytes to be determined, for example blood group specific
antigens, it is at present usual for antibodies to be provided in the indicator zones against
corresponding blood group antigens serving as bonding elements in successive or
superimposed bands in but a single flow track such as for example anti-A, anti-B against
the Rh blood group system. In this context conventional lateral flow tests suffer from the
disadvantage that the erythrocytes bonded to the antibodies form a flow barrier against the
analytes still to be tested for, for example further cell associated antigens, in a sample.
Due to agglutination or adsorption of cells in a band of bonding elements arranged
proximally to the application zone, additional analytes, in particular associated with cells
or cell fragments in the sample to be tested, can no longer be separated unimpededly and
visibly and can therefore not be tested for unambiguously or completely. For example in
a person who is blood group AB Rh D positive this may result in a weakening or
elimination of the B and the D bands, which may result in a faulty interpretation of being
blood group A Rh negative. For that reason it was hitherto not possible, specifically in
blood group serological diagnostics to employ a lateral flow test with more than one
indicator zone. In order to determine a plurality of, in particular cellular and plasmatic
blood group parameters, it is to date necessary to conduct single parameter tests
separately.
It is an object of the invention to overcome the disadvantages referred to of the state-of-
the-art, in particular of the successive or superposed indicator or testing zones of
conventional lateral flow tests for a simultaneous determination of different sample
parameters, in particular of cellular and plasmatic parameters.
The object is attained according to the invention on the one hand by a device for the
simultaneous qualitative or quantitative determination of one or more analytes in a liquid
sample or a plurality of liquid samples comprising a membrane with an application zone
for applying the liquid sample, at least two indicator zones which can enter into
interaction with the analyte(s) or with which analytes can interact and at least one
absorption region which takes up the liquid after having passed the indicator zones,
wherein the indicator zones are positioned between the applications zone and the
absorption region characterized in that the flow directions from the applications zone
through the respective indicator zones towards an absorption region, representing flow
tracts are substantially parallel, there being present at least two different flow tracks.
The indicator zones of the device according to the invention are present on the membrane
and comprise bonding elements which capture or bond the analyte(s) to be determined in
the sample. The bonding reactions between the analyte and the bonding element are
detected in the indicator zones.
In an embodiment of the invention the indicator zones are so arranged that the sample
liquid for each flow track will flow through not more than one indicator zone. For
example, the indicator zones are provided on the membrane in staggered relationship.
This arrangement of the indicator zones is, in this context, preferably configured in a row
extending diagonally from proximal to distal or vice versa. Particular embodiments are
V-shaped, W-, M- or N-shaped or reversed V-shaped W-, M- or N-shaped. In a further
embodiment the indicator zones are staggered parallel side by side in a linear row.
The provision of parallel staggered indicator zones is a precondition for a multi-parameter
testing with erythrocytes as indicator particles in a lateral arrangement. The particularly
preferred embodiment of a diagonal arrangement offers the advantage that the denotation
of the results can be applied to the device according to the invention in a particularly
practical and easily readable manner; because each parameter to be tested for occupies a
defined X and Y position the arrangement of the device according to the invention can be
considered as a coordinate system having an ordinate (plane of the direction of flow) and
an abscissar (plane of the application zone).
The indicator zones comprise antibodies or antibody fragments and/or lectines or
fragments thereof, which capture or bond to the blood antigens to be tested for and
thereby to the cells in the sample carrying those antigens. Antibodies or antibody
fragments and/or lectines or fragments thereof against antigens of all conceivable blood
group systems in die indicator zones are applied as preferred bonding elements in the
indicator zones on the porous membrane. Preferably a control bonding element (control =
ctl) which indicates the flow of the sample through the indicator zones positively, is
provided in an indicator zone, preferably in an indicator zone distally positioned in
relation to all remaining indicator zones. The control bonding element is preferably a
polyclonal anti-erythrocyte.
In a preferred embodiment it is provided that in one indicator zone a bonding element is
present, preferably an antibody or an antibody fragment against one analyte to be tested
for. Preferred embodiments of antibodies or antibody fragments and/or lectines or
fragments thereof in the indicator zones are antibodies or lectines against antigens of the
ABO blood group system, the Rh-, Kell-, Lewis- Hh, Duffy- Kidd, MNS-, Lutheran-, P-
systems. Further preferred as binding elements of the indicator zones are antibodies
against antigens of the blood group systems Diego, Yt, Scianna, Dombrock, Colton,
Chido/Rodgers, Gerbich, Cromer, Knops, Landsteiner-Wiener, Xg, Rx, India, Ok, Raph,
John Milton Hagen, Langereis, and/or Sid. A particularly preferred embodiment of the
device according to the invention includes indicator zones with the bonding elements
anti-A, -B, -AB, -D, -D, -C, -c, -E, -e, -Cw and/or -K antibodies or their antibody
fragments respectively wherein the two anti-D represent two different antibodies or their
antibody fragments. In particular, in the case of patients, pregnant women or new borns
these are preferably monoclonal antibodies of the IgM class which do not include the DVI
category. In the case of donors this is preferably an antibody which includes the D^
category and an antibody which does not embrace the D^ category.
Due to the device according to the invention it is no longer necessary for a blood group
determination to separately perform a pipetting for every single determination, instead,
for one sample, it is possible to simultaneously determine a large number of antigens of
interest for the blood group systems to be investigated, for example the most important
blood group characteristics of the blood group systems ABO, Rh and Kell (A, B, AB, D,
C, c, E, e, Cw, K). This represents an extraordinary rationalization of the procedures.
Likewise the reading of the results displayed in a diagonal pattern is substantially more
advantageous. Furthermore, with the device according to the invention it is possible to
determine and read side by side for example ABO- and Rh-properties in one device. The
allocation of the results to the respective patient is facilitated. The two-dimensional
planar result display as well as the stable end point of the reaction facilitate not only
reading with the naked eye but also an automatic reading of the results with conventional
display analysis procedures, such as e.g. CCD cameras. The work expenditure is reduced
even with manual processing. The device according to the invention moreover leads to a
reduction of environmental impact and to cost-effectiveness. Even in emergency
situations with time pressure it is possible to perform in a short period in a single test
setup, for example a complete ABO blood group/Rh-I intergroup determination. From a
production technological point of view the lateral diagonal flow design offers substantial
advantages over the state-of-the-art, in that a considerably reduced consumption of
reagents is achieved and also due to the provision of a multitude of test parameters in a
single device.
The device according to the invention provides a lateral flow test, in particular for blood
group serological diagnostics, wherein erythrocytes are used as indicator particles and
wherein in a single test batch simultaneously a plurality of cellular, in particular
erythrocytal antigens or antigen epitopes, plasmatic parameters and/or blood cell
properties can be determined.
Moreover, a test system is provided in this manner which can be produced in the simplest
possible manner and is easy to use, in particular with a small number of test runs and
without sample preparation and by means of which simultaneously a variety of cellular
parameters and/or plasmatic parameters of a sample or of a plurality of samples, in
particular blood group characteristics can be determined.
The membrane of the device according to the invention is a porous membrane. Preferred
membrane materials are, for example, nitrocellulose (e.g. uniS-art of Sartorius, HiFlow of
Millipore, Whatman, AE99 or FF85/100 of Schleicher & Schuell), polyethylene (Lateral
Flo of Porex Corporation) or nylon (Novalon of CUNO). Preferably the membrane has
the largest possible pore size because a high porocity of the membrane facilitates the
influx in particular of cellular components of the sample to be tested e.g. of erythrocytes
into the porous structure. The use of absorbent membranes is particularly advantageous.
However, the device according to the invention is not limited by such properties.
15Preferred are all membranes having a high capillary flow rate (capillary speed) wherein
the capillary flow rate represents that time which is required by a dye solution in order to
travel forty millimeters on a given membrane. Particularly preferred are membranes
having a capillary flow rate less than 100.
M a preferred embodiment of the invention a sealing element is provided on the porous
membrane downstream and upstream of the indicator zones of the application zone of the
device in accordance with the invention. Two- or three-dimensional sealing elements,
which are placed onto the porous membrane and by means of which a sample application
zone separate from the remaining surface of the porous membrane, are employed.
25According to the invention the sealing element primarily has the effect of a liquid barrier
and permits the directional distribution of sample liquid and test reagents in the porous
membrane. Moreover the sealing element according to the invention seals off the sample
application zone in order to prevent an inadvertent entry of liquid into the remaining parts
of the lateral flow device.
Preferred embodiments of the sealing element are web shapes or trough shapes or funnel
shapes. The shaping of the sealing element takes place by cutting processes from the
material used for the production of the sealing element. In the case of the funnel or
trough shape the sealing element is provided with an inner aperture, the preferred
modifications of which are round, square or rectangular and tapering towards the
underside (membrane contact side) of the sealing element in the case of the funnel shape.
Preferred materials for the sealing element are materials which are hydrophobic. In a
special embodiment, the materials are coated on one side with an adhesive film, for
example, a pressure sensitive or self-adhesive acrylate adhesive. Accordingly, the sealing
element can be adhesively bonded directly onto the surface of the porous membrane.
Alternatively, the sealing element can be bonded to the lateral flow casing, for example
adhesively bonded such that in this embodiment the lateral flow casing presses the sealing
element against the surface of the porous membrane such that the functions of the sealing
element are attained.
Preferred materials for the formation of two-dimensional sealing elements are any form of
adhesive tape or adhesive foils (e.g. Tesa 4124 of Beiersdorf AG, ARcare 7815 of
Adhesives Research).
Preferred materials for the formation of three-dimensional sealing elements are flexible,
closed pore elastomer materials or flexible silicon materials of variable material
thicknesses, preferably 3-5 mm (e.g. cellular caoutchouk EPDM140 of Pitzner, silicone
rubber or solid caoutchouk, hardness 40° or less of Castan).
Due to the structure according to the invention the device according to the invention is
capable of accommodating liquid samples which contain cells, for example complete
blood without filtering of the cells. Moreover, the sealing element permits the application
of large volumes of sample onto the porous membrane (application zone) without
fiooding thereof. Accordingly the sealing element supports the utilization of the
absorbent properties of the porous membrane. Furthermore, the sealing element ensures a
directional flow of sample. Nevertheless, the device according to the invention can
function well with or without any sealing element.
For the absorption region (absorption pad) of the device according to the invention,
mechanicaHy stable materials are preferred, preferably having a water absorption capacity
of 20-30 g/100 cm2 (e.g. Wicking Papier, type 300, Schleicher and SchuH). The contact
between the absorption pad and the lateral flow membrane of the device according to the
invention is produced by contact pressure and overlapping with the porous membrane.
The exact positioning of the absorption pad on the membrane is attained by adhesive
bonding of the absorption pad to the carrier layer (backing sheet) carrying the lateral flow
membrane.
In a further embodiment the components of the device according to the invention are
applied for purposes of mechanical strengthening onto a support or carrier layer. The
15device according to the invention can however also function without a carrier layer.
Preferably mechanically stable and non-water absorbent materials, preferably having
thicknesses of 100 um or more coated on one or both sides with an adhesive film e.g. a
pressure sensitive or self-adhesive acrylate adhesive (e.g. 0,005 inch polyester W7 GL-
187, G & L). On the carrier layer the porous membrane and the absorption pad are fixed.
In the case of a carrier layer rendered adhesive on both sides, the adhesive second side is
employed for fixing the stack onto further surfaces, e.g. inside the lateral flow casing.
In a further embodiment the device according to the invention, either with or without a
carrier layer, onto which the components of the device according to the invention have
been applied, is integrated in a casing, by which the membrane components are pressed
onto one another and the casing supports the sealing element function. However, in this
context the device according to the invention can function as well with as without a
casing.
A further subject of the invention is the use of the device according to the invention for
the analysis of blood, in particular for simultaneously performing the blood group
determination and serum cross-checking and/or antibody detection test and/or for the
simultaneous performance of the blood determination and the detection of antibodies
against infectious in particular bacterial and/or viral agents or fragments thereof or of
antigens of infective agents and/or for the simultaneous performance of blood group
determinations and the detection of antibodies against erythrocytes other than blood cells,
in particular anti-thrombocyte and/or anti-lymphocyte antibodies or fragments thereof.
A further subject of the invention is the use of the device according to the invention for
the analysis of blood, in particular simultaneous determination of blood group antigens or
antigen epitopes of any conceivable blood group system, preferably any conceivable
analytes on the surface of the red blood particles. The antigens or antigen epitopes to be
tested for are for example those of the ABO blood group system, the Rh-, Kell-, Lewis-
Hh-, Duffy- Kidd, MNS-, Lutheran-, P-system, the blood group systems Diego, Yt,
Scianna, Dombrock, Colton, Chido/Rodgers, Gerbich, Cromer, Knops, Landsteiner-
Wiener, Xg, Kx, India, Ok, Raph, John Milton Hagen, Langereis, and/or Sid, in particular
Al, A2, B, D, C, c E, e, Cw, K, k, M, N, S, s, Jk(a), Jk(b), Fy(a), Fy(b), Kp(a), Kp(b), Js
(a), Js(b), Le(a), Le(b), Lu(a), Lu(b), PI, I, H, Xg(a), U, Vw, Wr(a), Lan.
A preferred embodiment of the device according to the invention determines
simultaneously several blood group characteristics, for example A, B, AB, D, C, c, E, e,
Cw and K. The sample to be tested, for example native or anti-coagulated complete
blood or erythrocyte concentrates or diluted erythrocyte suspensions are applied onto the
application zone of the device according to the invention. The erythrocytes contained in
the sample which carry the analyte(s) serve simultaneously as indicator particles.
The object is attained according to the invention also by a process for determining a
plurality of analytes or their derivatives in a liquid sample comprising the application of
the sample onto the application zone of a membrane of the device according to the
invention, wherein the sample is present in adequate amount in order to induce the
sample liquid to flow in the direction of the absorption region through the indicator zones
and in order to induce the analytes or their derivatives in the sample liquid to become
bonded to the respective indicator zones or to form a complex in the indicator zones.
In the process according to the invention the analytes to be determined are in particular
blood group antigens or antigen epitopes of all blood group systems, preferably those
which are present on the surface of the red blood corpuscles. The antigens or antigen
epitopes to be tested for are for example those of the ABO blood group system, Rh-,
Kell-, Lewis- Hh-, Duffy- Kidd, MNS-, Lutheran-, P-systems, the blood group systems
Diego, Yt, Scianna, Dombrock, Colton, Chido/Rodgers, Gerbich, Cromer, Knops,
Landsteiner-Wiener, Xg, Kx, India, Ok, Raph, John Milton Hagen, Langereis, and/or Sid,
in particular Al, A2, B, D, C, c, E, e, Cw, K, k, M, N, S, s, Jk(a), Jk(b), Fy(a), Fy(b), Kp
(a), Kp(b), Js(a), Js(b), Le(a), Le(b), Lu(a), Lu(b), Lu(b), PI, I, H, Xg(a), U, Vw, Wr(a),
Lan.
A preferred embodiment of the process according to the invention identifies
simultaneously a plurality of blood group characteristics, for example A, B, AB, D, C, c,
E, e, Cw and K. The sample to be tested, for example native or anti-coagulated complete
blood or erythrocytes suspensions with or without test liquid, such as control blood is
applied onto the application zone of the device according to the invention. The
erythrocytes contained in the sample which carry the analyte(s) simultaneously serve as
indicator particles.
BRIF DESCRIPTION OF THE ACCOMPANYING DRAWINGS
In the following the invention will be further illucidated by figures and examples without
being limited thereby. There is shown in:
Fig. 1 a perspective view of a device according to the invention for lateral flow tests for
the simultaneous determination of blood group characteristics A, B, AB, D and CDE;
Fig. 2 an explosive view of the device for lateral flow tests according to the invention
illustrated in Fig. 1;
Fig. 3 a perspective view of a device according to the invention for lateral flow tests for
the simultaneous performance of determination of blood group characteristics A, B, AB,
D and CDE carried out with a three-dimensional sealing element in the form of a web;
Fig. 4 an explosive view of the device according to the invention for lateral flow tests
illustrated in Fig. 3;
Fig. 5 a perspective view of a device according to the invention for lateral flow tests for
the simultaneous determination of blood group characteristics performed with a three-
dimensional sealing element in the form of a trough;
Fig. 6 an explosive view of the device for lateral flow tests according to the invention
illustrated in Fig. 5;
Fig. 7 a perspective view of a device according to the invention for lateral flow tests for
the simultaneous determination of blood group characteristics A, B, AD, C, c, E, e, Cw
and K;
Fig. 8 a perspective view of a device according to the invention for lateral flow tests
performed as bedside tests for testing the ABO identity of the recipient and the blood
preparation.
Fig. 9 an exploded view of the device according to the invention for lateral flow tests
illustrated in Fig. 8.
Fig. 10 a perspective view of a device according to the invention for lateral flow tests for
the simultaneous determination of blood group characteristics A, B, AB, D and CDE.
Fig. 11a perspective view of a device according to the invention for lateral flow tests for
the simultaneous determination of blood group characteristics A, B, AB, D and CDE.
Fig. 12. a perspective view of a device according to the invention for lateral flow tests for
the simultaneous determination of blood group characteristics A, B, AB, D and CDE.
Fig. 13 a perspective view of a device according to the invention for lateral flow tests for
the simultaneous determination of blood group characteristics A, B, AB, D and CDE.
Fig. 14 a perspective view of a device according to the invention for bi-directional lateral
flow tests for the simultaneous determination of blood group characteristics A, B, AB, D
and CDE.
Fig. 15 an exploded view of the device according to the invention for lateral flow tests as
illustrated in Fig. 14.
Fig. 1 shows by way of example a perspective illustration of a device according to the
invention for lateral flow tests for the simultaneous performance determination of blood
group A, B, AB, D and CDE. In the present example the device comprises a support
layer 1, the porous membrane 2 the absorption pad 3 and the two-dimensional sealing
element 4 in the form of a strip. The porous membrane 2 is fixed onto the support layer 1
by means of a pressure sensitive acrylic adhesive. Likewise, the absorption pad 3 is fixed
onto the support layer 1, part of the absorption pad 3 overlapping the porous membrane 2.
The sealing element 4 fixed on the upper side of the porous membrane 2 separates the
application zone 5 from the remaining membrane surface and permits the directed
distribution of sample liquid and test reagents into the porous membrane 2. Between the
application zone 5 and the region of the porous membrane 2 which is in contact with the
absorption pad 3 the indicator zone region 6 is provided. The latter is formed by
diagonally staggered point-shaped indicator zones I-VI arranged in defined X and Y
positions wherein the indicator zones are composed of the following bonding elements:
Indicator zone VI is the control (ctl) and contains polyclonal anti-erythrocyte antibodies.
They are arranged in distal relationship to all remaining indicator zones.
In Fig. 2 an explosive view of the device according to the invention for lateral flow tests
illustrated in Fig. 1 is shown comprising the components support layer 1, porous
membrane 2, absorption pad 3 and sealing element 4 which separate the application zone
5 from the remainder of the membrane which in turn comprises the indicator zone region
including the indicator zones I-IV diagonally staggered from proximal to distal.
In Fig. 3 a perspective view of a device according to the invention for lateral flow tests
for the simultaneous determination of blood group characteristics A, B, AD, D and CDE
is shown by way of example. In the present example the components of the device
correspond to the components of the device illustrated in Fig. 1 except for the sealing
element 4 fixed to the upper side of the porous membrane 2 in the form of a three-
dimensional batten.
In Fig. 4 an explosive view is shown of the device according to the invention illustrated in
Fig. 3 for lateral flow tests including the components support layer 1, porous membrane
2, absorption pad 3 and a sealing element 4 in the form of a three-dimensional batten
which separates the application zone from the remaining membrane which in turn
contains the indicator zone region 6 comprising the indicator zone regions determination
including the indicator zones I-VI diagonally staggered from proximal to distal.
In Fig. 5 is shown by way of example a perspective illustration of a device according to
the invention for lateral flow tests for the simultaneous determination of blood group
characteristics A, B, AB, D and CDE. In the present example the components of the
device correspond to the components of the device as illustrated in Fig. 1 except for the
sealing element 4 fixed to the porous membrane 2 in the form of a three-dimensional
trough.
In Fig. 6 an exploded view of the device according to the invention for lateral flow tests
illustrated in Fig. 5 is shown including the components support layer 1, porous membrane
2, absorption pad 3 and sealing element 4 in three-dimensional trough form which
separates the application zone 5 from the remainder of the membrane which in turn
consists of the indicator zone region 6, including the indicator zones I-V diagonally
staggered from proximal to distal.
Fig. 7 illustrates by way of example a perspective view of a device according to the
invention for lateral flow tests for the simultaneous determination of blood group
characteristics A, B, AB, D, C, c, E, e, Cw and K In the present case the device is
composed of a support layer 1, the porous membrane 2, the absorption pad 3 and the two-
dimensional sealing element 4 in the form of a strip. The membrane 2 is affixed on the
support layer 1 provided with a pressure sensitive acrylic adhesive. Likewise the
absorption pad 3 is affixed to the support layer 1 with part of the absorption pad 3
overlapping the porous membrane 2. The sealing element 4 fixed to the upper side of the
porous membrane 2 separates the application zone 5 from the remaining membrane
surface and permits the directional distribution of sample liquid and test reagents into the
porous membrane 2. Between the application zone 5 and the region of the porous
membrane 2 which is in contact with the absorption pad 3, the indicator zone region 6 is
provided. The latter is formed by diagonally staggered point-shaped indicator zones 1-XI
provided in defined X and Y positions, consisting of the following binding elements:

Indicator zone XI is the control (ctl) and contains polyclonol anti-erythrocyte antibodies.
They are arranged in distal relationship to all remaining indicator zones.
In Fig. 8 there is illustrated by way of example a perspective view of a device according
to the invention for lateral flow tests, designed as a bedside test for testing the ABO
identity of the recipient and the blood preserve. In the present example the device
comprises a support layer 1, the porous membranes 2a and 2b present in dual form, the
absorption pad 3 and the two-dimensional sealing elements 4a and 4b presented in strip
form. The two porous membranes 2a and 2b are fixed by way of a support layer 1
provided with a pressure sensitive or acrylate adhesive parallel to one another and
orientated in the same direction. Likewise the absorption pad 3 is fixed onto the support
layer 1, part of the absorption pad 3 overlapping equidistantly with both porous
membranes 2a and 2b. The sealing elements 4a and 4b fixed onto the upper side of the
porous membranes 2a and 2b separate the respective application zones 5 a and 5b from the
remaining membrane surface and permit the directed distribution of sample liquid and
test reagents in the porous membranes 2a and 2b. Between the application zone 5a or 5b
respectively and the respective regions of the porous membranes 2a and 2b, which are in
contact with the absorption pad 3, the indicator zone regions 6a and 6b are provided.
These are formed by point-shaped indicator zones Ia-IHa and Ib-IHb respectively in
defined X and Y positions, the indicator zones being composed of the following bonding
elements.

Indicator zones IIIa and IIIb are the controls (ctl) and contain polyclonal anti-erythrocyte
antibodies. They are provided in distal relationship to all remaining indicator zones.
In Fig. 9 there is illustrated an exploded view of the device according to the invention for
lateral flow tests including the components support layer I, porous membranes 2a and 2b,
absorption pad 3 and the sealing elements 4a and 4b as shown in Fig. 8, the latter each
separating the application zones 5 a and 5b respectively from the remainder of the
membrane which in turn contains the indicator zone regions 6a and 6b respectively
including the indicator zones Ia-IHa and Ib-IHb provided in diagonal staggered
relationship from proximal to distal.
In Fig. 10 there is shown by way of example a perspective view of a device according to
the invention for lateral flow tests for the simultaneous determination of the blood group
characteristics A, B, AB, D and CDE. The present example represents a lateral flow test
device for right-handed operators and comprises a support layer 1, the porous membrane
2, the sealing element 4. The porous membrane 2 is fixed onto the support layer 1
provided with a pressure sensitive or self-adhesive acrylate adhesive. Likewise the
absorption pad 3 is fixed on the support layer 1, part of the absorption pad 3 overlapping
with the porous membrane 2. The sealing element 4 fixed onto the upper side of the
porous membrane 2 separates the application zone 5 from the remainder of the membrane
surface and permits a directional distribution of sample liquid and test reagents in the
porous membrane 2. The indicator zone region 6 is provided between the application
zone 5 and the region of the porous membrane 2 which is in contact with the absorption
pad 3. This indicator region is formed by point-shaped indicator zones I-VI arranged
parallel side by side in staggered relationship in a linear row in defined X and Y
positions, wherein the indicator zones are composed of the following bonding elements:

Indicator zone VI represents the control (ctl) and contains polyclonal anti-erythrocyte
antibodies. It is arranged in distal relationship to all remaining indicator zones.
In Fig. 11 there is shown by way of example a perspective view of a device according to
the invention for lateral flow tests for the simultaneous determination of the blood group
characteristics A, B, AB, D and CDE. The present example represents a lateral flow test
device for left-handed operators and comprises a support layer 1, the porous membrane 2,
the absorption pad 3 and the two-dimensional sealing element 4 in strip form. The porous
membrane 2 is fixed onto the support layer 1 provided with a pressure sensitive or self-
adhesive acrylate adhesive. Likewise the absorption pad 3 is fixed onto the support layer
1, part of the absorption pad 3 overlapping with the porous membrane 2. The sealing
element 4 fixed onto the upper side of the porous membrane 2 separates the application
zone 5 from the remainder of the membrane surface and permits the directional spreading
of the sample liquid and the test reagents in the porous membrane 2. Between the
application zone 5 and the region of the porous membrane 2 which is in contact with the
absorption pad 3 the indicator zone region 6 is provided. The latter is formed by point-
shaped indicator zones 1-VI, arranged parallel staggered side by side in a linear row in
defined X and Y positions, the indicator zones being composed of the following bonding
elements:

Indicator zone 6 is the control (ctl) and contains polyclonal anti-erythrocyte antibodies. It
is provided in distal relationship to all remaining indicator zones.
In Fig. 12 there is shown by way of example a perspective view of a device according to
the invention for lateral flow tests for the simultaneous determination of the blood group
characteristic A, B, AB, D and CDE. The present example represents a lateral flow test
device for right-handed operators and comprises a support layer 1, the porous membrane
2, the absorption pad 3 and the two-dimensional sealing element 4 in the form of a strip.
The porous membrane 2 is fixed onto the support layer 1 by means of a pressure sensitive
or self-adhesive acrylate adhesive. Likewise the absorption pad 3 is fixed onto the
support layer 1 such that a portion of the absorption pad 3 overlaps with the porous
membrane 2. The sealing element 4 fixed onto the upper side of the porous membrane 2
separates the application zone 5 from the remaining membrane surface and permits the
directional spreading of sample liquid and test reagents into the porous membrane 2. The
indicator zone 6 is arranged between the application zone 5 and the region of the porous
membrane 2 which is in contact with the absorption pad 3. This indicator zone region is
formed by elongate or band-shaped indicator zones I-VI staggered side by side parallel to
one another in defined X and Y positions, the indicator zones being composed of the
following bonding elements:

Indicator zone VI is the control (ctl) and contains polyclonal anti-erythrocyte antibodies.
It is provided in distal relationship to all remaining indicator zones.
In Fig. 13 there is shown by way of example a perspective view of a device according to
the invention for lateral flow tests for the simultaneous determination of blood group
characteristics A, B, AB, D and CDE. The present example represents a lateral flow test
device for left-handed operators and comprises a support layer 1, the porous membrane 2,
the absorption pad 3 and the two-dimensional sealing elements 4 provided in a strip form.
The porous membrane 2 is fixed onto the support layer 1 provided with a pressure
sensitive or self-adhesive acrylate adhesive. Likewise the absorption pad 3 is fixed onto
the support layer 1, part of the absorption pad 3 overlapping with the porous membrane 2.
The sealing element 4 fixed onto the upper side of the porous membrane 2 separates the
application zone 5 from the remaining membrane surface and permits the directed
spreading of sample liquid and test reagents into the porous membrane 2. Between the
application zone 5 and the region of the porous membrane 2 which is in contact with the
absorption pad 3, an indicator zone 6 is provided. The latter is formed by elongate or
band-shaped indicator zones I-VI arranged parallel side by side in staggered relationship
in defined X and Y positions, the indicator zones being composed of the following
bonding elements:

Indicator zone VI is the control (ctl) and contains polyclonal and anti-erythrocyte
antibodies. It is arranged in distal relationship to all remaining indicator zones.
In Fig. 14 there is shown by way of example a perspective view of a device according to
the invention for lateral flow tests with bi-directional flow for the simultaneous
determination of the blood group characteristics A, B, AB, D and CDE. In the present
example the device comprises a support layer 1, the porous membrane 2, the absorption
pads 3a and 3b and the two-dimensional sealing elements 4a and 4b provided in strip
form. The porous membrane 2 is fixed onto the support layer 1 by means of a support
layer provided with a pressure sensitive acrylate adhesive. Likewise the absorption pads
3a and 3b are fixed onto the support layer 1, a portion of the absorption pads 3a and 3b
overlapping with the porous membrane 2. The sealing elements 4a and 4b fixed onto the
upper side of the porous membrane 2 separate the application zone 5 provided in the
central region of the membrane from the remaining membrane surface and permit the bi-
directional distribution of sample liquid and test reagents in the porous membrane 2.
Between the application zone 5 and the region of the porous membrane 2 which is in
contact with the absorption pads 3a and 3b, the indicator zone regions 6a and 6b are
provided. These are represented by point-shaped indicator zones I-VI arranged diagonally
staggered in defined X and Y positions, the indicator zones being composed of the
following bonding elements:

The indicator zone VIa and VIb are the controls (ctl) and contain polyclonal anti-
erythrocyte antibodies. They are distally arranged in relation to the indicator zones I-III
and IV-V respectively.
In Fig. 15 there is shown an exploded view of the device according to the invention
illustrated in Fig. 14 for lateral flow tests with bi-directional flow comprising the
components support layer 1, porous membrane 2, absorption pads 3a and 3b and the
sealing elements 4a and 4b which separate the centrally disposed application zone 5 from
the remaining membrane surface which in turn contains two indicator zone regions 6a and
6b including the indicator zones I, II, II, VIa and IV, V, VIb respectively which are
arranged diagonally staggered from proximal to distal.
Examples
Example 1: Blood group determinations
Production of test strips:
The test strips comprise an application zone, an indicator zone region and an absorption
region. Membranes of the type Millipore HiFlow Plus 065 are cut to size in strips, sized
15 by 35 mm (width/length; x/y) for a six spot embodiment or respectively sized 26 by
40 mm for an 11 spot embodiment and adhesively fixed onto a support layer (backing
sheet e.g. of G&L). Diagonally or alternatively staggered in a linear row 0,2 ul dots of
solutions of various blood groups specific monoclonal antibodies are applied in the
indicator zone region using a dispenser, e.g. AD3200 (Biodot):
Anti-A clone Birma-1 (Serologicals, TLJ0105); anti-B clone ES-4 (Serologicals,
NCA0201); anti-AB clones AB6, AB26, AB92 (Medion Diagnostics, 010062); anti-D
clone LDM3 (SNBTS, Z7180100); anti-C clone MS-24 (Serologicals, unformulated,
KGK0212; Anti-c clone MS-33 (Seriologicals, KNI0207); anti-E clones MS-80 + MS-
258 (Serologicals, KXE0201); clones anti-e MS-21+MS-63 (Serologicals,
KLL0205+KQK0205); anti-Cw clone MS-110 (Serologicals, JPK0201); anti-K - clone
MS-56, (Serologicals, KOA0201).
The positioning of the anti-A antibodies takes place in position x=3/y=10 mm. All other
antibodies are dispensed iterating at distances of x=l,5/y=2,2 mm in relation to the
position of the anti-A antibody. The anti-erythrocytes specific control antibody (rabbit
IgG fraction of anti-human RBC, Rockland, 209-4139) are applied in x=2/y=3,5 mm
staggered in relation to the last spot of the series of blood groups specific antibodies. The
dilutions of the antibodies is performed in 15 mM potassium phosphate buffer pH 7,5, 10
% (v/v) methanol as follows: anti-A antibody 1:3, anti-B antibody 1:2, anti-AB antibody
1:4, anti-D antibody 1:4, anti-RBC antibody 1:3. All other antibody solutions are not pre-
diluted but are mixed with methanol to 10 % (v/v).
The membranes after the dispensing of the antibodies are dried for 20 minutes at 40°C
and subsequently stored at constant air humidity until the test is performed. At the end
distal to the application zone an absorption pad sized 15x10 mm or 26x10 mm as the case
may be (Schleicher & Schull, 300) is adhesively applied overlapping with the membrane
by 3 mm. The application zone is separated from the remainder of the membrane by the
adhesive application of a 1-2 mm wide adhesive strip (Tesa 4124) in position y=5 mm
extending over the entire width of the membrane.
Test batch:
Anti-coagulated full blood batches are used for the blood tests. For the test proper 100 ul
of diluted blood in a ratio of 1:6 blood to dilution buffer (Enlisstll, Medion Diagnostics or
diluent 1, DiaMed) (6 spot embodiment) or 150 ul (11 splot embodiment) are applied in
the application zone. Once the blood has left the application zone 100 ul or 150 ul
dilution buffer or preferably lµl hypo-osmotic rinsing buffer (15 mM potassium
phosphate buffer pH 7,4, 0,3-0,45 % (w/v) NaCl) are pipetted onto the application zone in
order to rinse unbound erythrocytes from the membrane. However, in the alternative, the
sample application may be performed with 50 ul 1:3 diluted or undiluted blood. With
those samples the membrane is rinsed twice with dilution buffer or once with dilution
buffer followed by hypo-osmotic rinsing buffer.
If the 1:6 dilution is selected the anti-RBC control as indicator of a successfully
performed test is visible after 2 minutes. With undiluted blood the test takes longer.
Result:
The test is valid if the anti-RBC control displays a clearly positive signal (red dot).
Depending on the presence or absence of the respective blood group antigens red dots
(positive) or the almost white background colouration of the membrane (negative) appear
in the respective positions.
Example 2: Bedside test
Production of test strip:
The bedside test comprises two membranes each ("blood preserve", "recipient") fixed
onto a support layer (backing sheet), each comprising an application zone, an indicator
zone region and an absorption region.
Membranes of the type Millipore HiFlow Plus 065 are cut to size in strips of a size 12,5 x
30 mm (width/length); x/y). Two each thereof are adhesively bonded onto a support layer
(backing sheet e.g. of G&L) spaced apart by 5 mm so that the overall assembly has a size
of 30 x 30 mm.
Using a dispenser, e.g. AD3200 (Biodot) the following identical applications are applied,
staggered diagonally onto each of the two membranes: 0,2 ul dots of solutions of the
monoclonal antibody anti-A clone Birma-1 (Serologicals, TLJ0105) in position x=4/y=12
mm; anti-B-clone ES-4 (Serologicals, NCA0201) in position x=7/y=14 mm. The anti-
erythrocyte specific control antibody (rabbit IgG fraction of anti-human RBC, Rockland,
209-4139) is applied in staggered relationship x=3/y=4 mm to the anti-B spot. The
dilutions of the antibodies are performed in 15 mM potassium phosphate buffer pH 7,5,
10 % (v/v) methanol as follows: anti-A antibody 1:3, anti-B antibody 1:2, anti-RBC
antibody 1:3.
The membranes, after dispensing the antibody, are dried for 20 minutes at 40°C and are
stored at constant air humidity until the test is performed. At the end distal to the
application zone an absorption pad (Schleicher & Schiill, 300) sized 30x10 mm is
applied, overlapping with both membranes by 3 mm. The application zone is separated
from the remainder of the membrane by the adhesive application of a 1-2 mm wide for
each test strip (Tesa 4124) in position y=5 mm extending over the entire membrane
width.
Test batch:
Serving as samples there are used: for the membrane "preserve": erythrocyte concentrate;
for the membrane "recipient": complete blood.
For the test proper 50 ul complete blood is applied in the respective application zone on
the side "recipient" and 50 ul erythrocyte concentrate on the side "preserve". After the
blood has been sucked in by the membrane entirely, rinsing is performed in each case
with 2 x 100 ul dilution buffer or once with dilution buffer following by hypo-osmotic
rinsing buffer.
Result:
The anti-RBC control as indicator of a successfully performed test becomes visible in
both membranes after about 2 minutes.
The test is valid if the anti-RBC control displays a clearly positive signal (red dot).
Depending on the presence or absence of the respective blood group antigens red dots
(positive) or the almost white background colouration of the membrane (negative) are
displayed at the respective positions. An identical display for "recipient" and "preserve"
denotes ABO identity between the recipient and the preserve.
Example 5: Blood group determination with bi-directional lateral flow test
Production of the test strips:
The test strips comprise an application zone in the central region of the membrane, two
indicator zone regions and two absorption regions. Membranes of the type Millipore
HiFlow Plus 065 are cut to size in dimensions of 15 x 50 mm (width/length, x/y) and
adhesively bonded onto a support layer (backing sheet e.g. G&L). Diagonally staggered
or alternatively in a linear row staggered, 0,2 ul dots of solutions of different blood group
specific monoclonal antibodies are applied in the indicator zone region. The central
region of the test strip (y=0 mm) serves as the reference dimension for the positioning of
the indicator zones in the y direction. The following antibodies are dispensed using a
dispenser, e.g. AD3200 (Biodot):
Anti-a-clone Birma-1 (Serologicals, TLJ0105); anti-B-clone ES-4 (Serologicals,
NCA0201); anti-AB-clones AB6, AB26, AB92 (Medion Diagnostics, 010062); anti-D-
clone LDM3 (SNBTS, Z7180100); anti-C-clone MS-24 (Serologicals, unformulated,
KGK0212); anti-E-clones MS-80 + MS-258 (Serologicals KXE0201). For the anti-CDE
indicator zones the anti-D and anti-C antibodies are concentrated two-fold the anti-E
antibodies three-fold and mixed in equal proportions by volume.
In the embodiment of diagonal staggering of the indicator zones, the dispensing of the
anti-A antibody is performed in position x=4/y=10 mm. The positions of the anti-B and
anti-AB antibodies proceed iterating at distances of x=l,5/y=2 mm in relation to the
position of the anti-A antibody. The anti-erythrocyte specific control antibody (rabbit IgG
fraction of anti-human RBC, Rockland, 209-4139) is applied at x=3,5/y=3,5 mm
staggering in relation to the last spot of the series of the anti-A, anti-B and anti-AB
antibodies. The dispensing of the anti-D antibody takes place in position x=4/y=10 mm.
The dispensing of the anti-CDE antibody at a distance of x=3,5/y=2 mm. The anti-
erythrocyte specific control antibody is applied at x=3,5/y=3,5 mm staggering in relation
to the spot of the anti-CDE antibody. The dilution of the antibodies proceeds in 15 mM
potassium phosphate buffer pH 7,5, 10% (v/v) methanol as follows: anti-A antibody 1:3,
anti-B antibody 1:2, anti-AB antibody 1:4, anti-D antibody 1:3 and anti-RBC antibody
1:3. The antibody mixture anti-CDE is not pre-diluted but is mixed with methanol to 10
% (v/v) concentration.
The membranes, after the dispensing of the antibodies, are dried for 20 minutes at 40°C
and subsequently stored at constant air humidity until the test is performed. At the end of
the membrane, distal to the application zone, two absorption pads (Schleicher & Schtill,
300) sized 15x10 mm are bonded adhesively onto the membrane overlapping by 3 mm.
The application zone is separated over the entire width of the membrane from the
remainder of the membrane by two 1-2 mm wide test strips (Tesa 4124) in position y=3
mm or y=3mm respectively.
Test batch:
Anti-coagulate complete blood batches are used as blood samples. For the test proper
100 ul of diluted blood in a ratio of 1:6 in dilution buffer (EnlisstE, Medion Diagnostics)
is applied in the application zone. Once the blood has left the application zone a single
application of 100 ul dilution buffer or of 100 ul hypo-osmotic rinsing buffer (15 mM
potassium phosphate buffer pH 7,4, 0,3-0, 45 % (w/v) NaCl) is pipetted onto the
application zone in order to rinse non-bonded erythrocytes from the membrane.
Alternatively, the sample application may proceed with 50 ul 1:3 diluted or undiluted
blood. In the case of those samples the membrane is rinsed twice with dilution buffer or
once with dilution buffer and thereafter with hypo-osmotic rinsing buffer.
In the case of the 1:6 dilution being selected, the anti-RBC control as indicator of a
successfully performed test is visible after 2 minutes. With undiluted blood the test takes
longer.
Result:
The test is valid, if the anti-RBC control displays a clearly visible signal (red dot).
Depending on the presence or absence of the respective blood group antigens, there are
displayed in the corresponding positions red dots (positive) or the almost white
background colouration of the membrane (negative).
WE CLAIM:
1. Device for the simultaneous and qualitative or quantitative determination of a plurality of
analytes in a liquid sample such as herein described, comprising a membrane (2) with
- an application zone (5) for the application of the liquid sample,
- at least one group of at least two indicator zones, which are able to interact with the analyte(s)
and
- at least one absorption region (3) which takes up the liquid after having passed the indicator
zones
wherein the indicator zones are located between the application zone (5) and the absorption
region (3), characterized in that the flow directions from the application zone (5) through the respective
indicator zones of a group towards an absorption region (3) (flow tracks) are substantially parallel and
that at least two different flow tracks are present, and wherein the indicator zones are so arranged that
the test liquids for any one flow track flow through not more than one indicator zone.
2. Device as claimed in claim 1, wherein the indicator zones are arranged in a diagonal V-, W-,
M-, N-shaped or linear row.
3. Device according any one of claims 1 to 2, wherein the indicator zones comprise antibodies or
antibody fragments or lectines, antigens or antigen epitopes and/or cells or cell fragments.
4. Device as claimed in any one of claims 1 to 3, wherein the indicator zones comprise in
particular anti-A, B, -HB, -D, -D, -C, -c, E, -e, -Cw and/or Kantibodies or antibody fragments.
5. Device as claimed in any one of claims 1 to 4, wherein all the membranes (2) preferably consist
of polyethylene, nitrocellulose or nylon.
6. Device as claimed in any one of claims 1 to 5, wherein downstream of the application zone (5)
and upstream of the indicator zones at least one sealing element (4) is provided on the membrane (2).
7. Device as claimed in any one of claims 1 to 6, wherein the components of the device have been
applied onto a support layer (1) for mechanical reinforcement.
8. Device as claimed in any one of claims 1 to 7, wherein the components of the device are
integrated in a casing.
9. A device as claimed in any one of claims 1 to 8 for the analysis of blood, in particular for the
determination of blood group antigens or antigen epitopes.
10. A device as claimed in any one of claims 1 to 9 for the analysis of blood, in particular for the
simultaneous determination of A-, B-, AB-, D-, C, c-, E-, e, Cw - and/or K-blood group antigens or
antigen epitopes.
11. Method for the determination of a plurality of analytes in a liquid sample such as herein
described, comprising:
the application of the sample onto the application zone (5) of a membrane (2) of the device as
claimed in any one of the preceding claims 1 to 8, wherein this sample is present in adequate amounts
in order to induce the test liquid to flow in the direction of the absorption region (3) through the
indicator zones and to induce the analytes or their derivatives in the test liquid to form a complex in the
indicator zones.
12. Method as claimed in claim 11, wherein the analytes are blood group antigens or antigen
epitopes.
13. Method as claimed in claim 11 or 12, wherein the analytes in particular include A-, B-, AB-, D-,
C, c-, E-, e, Cw - and/or K-blood group antigens or antigen epitopes.
14. Method as claimed in any one of claims 11 to 13, wherein the analytes A-, B-, AB-, D-, C, c-,
E-, e, Cw - and/or K-blood group antigens or antigen epitopes are detected simultaneously.
15. Method as claimed in any one of claims 11 to 14, wherein the indicator particles are
erythrocytes.
16. Method as claimed in any one of claims 11 to 15, wherein the membrane (2) after the!
application of indicator particles is rinsed.
17. Method as claimed in claim 16 wherein the rinsing liquor is preferably hypo-osmotic. I
18. Method as claimed in any one of claims 11 to 17, wherein the liquid sample is composed of
blood or blood components, preferably of complete blood, erythrocyte concentrate or test liquid such as
control blood.


Device for the simultaneous and qualitative or quantitative determination of a plurality of
analytes in a liquid sample, comprising a membrane 2 with
5 - an application zone 5 for the application of the liquid sample,
at least one group of at least two indicator zones, which are able to interact
with the analyte(s) and
at least one absorption region 3 which takes up the liquid after having
passed the indicator zones
10 wherein the indicator zones are located between the application zone 5 and
the absorption region 3, characterized in that
the flow directions from the application zone 5 through the respective indicator
zones of a group towards an absorption region 3 (flow tracks) are substantially
parallel and that at least two different flow tracks are present.
15
Furthermore, the invention relates to a method for the determination of a plurality of
analytes or their derivatives in a liquid sample, comprising:
the application of the sample onto the application zone 5 of a membrane 2 of the
device according to any one of the preceding claims 1 to 8, wherein this sample is
20 present in adequate amounts in order to induce the test liquid to flow in the
direction of the absorption region 3 through the indicator zones and to induce the
analytes or their derivatives in the test liquid to form a complex in the indicator
zones.

Documents:

00204-kolnp-2006-abstract.pdf

00204-kolnp-2006-claims.pdf

00204-kolnp-2006-description complete.pdf

00204-kolnp-2006-drawings.pdf

00204-kolnp-2006-form 1.pdf

00204-kolnp-2006-form 3.pdf

00204-kolnp-2006-form 5.pdf

00204-kolnp-2006-international publication.pdf

00204-kolnp-2006-international search authority.pdf

00204-kolnp-2006-pct forms.pdf

00204-kolnp-2006-priority document.pdf

204-KOLNP-2006-ABSTRACT 1.1.pdf

204-KOLNP-2006-ABSTRACT 1.2.pdf

204-KOLNP-2006-ANNEXURE FORM 3.pdf

204-kolnp-2006-assignment 1.2.pdf

204-KOLNP-2006-ASSIGNMENT.pdf

204-KOLNP-2006-CANCELLED PAGES.pdf

204-KOLNP-2006-CLAIMS 1.1.pdf

204-KOLNP-2006-CLAIMS 1.2.pdf

204-kolnp-2006-correspondence 1.2.pdf

204-KOLNP-2006-CORRESPONDENCE.pdf

204-KOLNP-2006-DESCRIPTION (COMPLETE) 1.1.pdf

204-KOLNP-2006-DESCRIPTION (COMPLETE) 1.2.pdf

204-KOLNP-2006-DRAWINGS 1.1.pdf

204-kolnp-2006-examination report.pdf

204-KOLNP-2006-FORM 1.1.1.pdf

204-KOLNP-2006-FORM 1.1.2.pdf

204-kolnp-2006-form 13 1.1.pdf

204-KOLNP-2006-FORM 13.pdf

204-kolnp-2006-form 18 1.1.pdf

204-KOLNP-2006-FORM 3.1.1.pdf

204-KOLNP-2006-FORM 5.1.1.pdf

204-KOLNP-2006-FORM 6.pdf

204-KOLNP-2006-FORM-27.pdf

204-kolnp-2006-gpa.pdf

204-kolnp-2006-granted form 3.pdf

204-kolnp-2006-granted form 5.pdf

204-kolnp-2006-granted-abstract.pdf

204-kolnp-2006-granted-claims.pdf

204-kolnp-2006-granted-description (complete).pdf

204-kolnp-2006-granted-drawings.pdf

204-kolnp-2006-granted-form 1.pdf

204-kolnp-2006-granted-specification.pdf

204-KOLNP-2006-OTHERS 1.1.pdf

204-KOLNP-2006-OTHERS.pdf

204-KOLNP-2006-PA.pdf

204-KOLNP-2006-PETITION UNDER RULE 137.pdf

204-KOLNP-2006-REPLY TO EXAMINATION REPORT 1.1.pdf

204-kolnp-2006-reply to examination report 1.2.pdf

204-KOLNP-2006-REPLY TO EXAMINATION REPORT.pdf

204-KOLNP-2006-TRANSLATED COPY OF PRIORITY DOCUMENT.pdf

abstract-00204-kolnp-2006.jpg


Patent Number 243402
Indian Patent Application Number 204/KOLNP/2006
PG Journal Number 42/2010
Publication Date 15-Oct-2010
Grant Date 13-Oct-2010
Date of Filing 27-Jan-2006
Name of Patentee MEDION DIAGNOSTICS AG
Applicant Address BONNSTRASSE 9, 3186 DUEDINGEN
Inventors:
# Inventor's Name Inventor's Address
1 SCHWIND PETER CHEMIN DU CALVAIRE 4, 1700 FRIBOURG
2 LOSTER KLEMENS SIEGELSTRASSE 4, 16562 BERGFELDE
PCT International Classification Number G01N 33/80
PCT International Application Number PCT/EP2004/007536
PCT International Filing date 2004-07-08
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 103 30 982.9 2003-07-09 Germany