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Patent 2570383 Summary

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(12) Patent: (11) CA 2570383
(54) English Title: FILTER DEVICE, THE METHOD, KIT AND USE THEREOF
(54) French Title: DISPOSITIF DE FILTRE, PROCEDE, KIT ET UTILISATION DE CELUI-CI
Status: Deemed expired
Bibliographic Data
(51) International Patent Classification (IPC):
  • G01N 33/543 (2006.01)
  • G01N 33/58 (2006.01)
(72) Inventors :
  • SARAMAKI, MIKA (Finland)
  • NISKANEN, AIMO (Finland)
(73) Owners :
  • ANI BIOTECH OY (Finland)
(71) Applicants :
  • ANI BIOTECH OY (Finland)
(74) Agent: BERESKIN & PARR LLP/S.E.N.C.R.L.,S.R.L.
(74) Associate agent:
(45) Issued: 2015-04-14
(86) PCT Filing Date: 2005-06-15
(87) Open to Public Inspection: 2005-12-29
Examination requested: 2010-06-11
Availability of licence: N/A
(25) Language of filing: English

Patent Cooperation Treaty (PCT): Yes
(86) PCT Filing Number: PCT/FI2005/050215
(87) International Publication Number: WO2005/124347
(85) National Entry: 2006-12-14

(30) Application Priority Data:
Application No. Country/Territory Date
20040825 Finland 2004-06-15
10/954,627 United States of America 2004-09-29

Abstracts

English Abstract




The invention provides an immunochemical filter device and use thereof, said
filter device comprising a filter material attached to a support member, for
example a cap of a vessel. The filter material comprises a labeled specific
binding reagent, wherein said labeled specific binding reagent is activated by
filtration of a liquid sample solution through the filter material. The
mixture of the sample solution and the labeled specific binding reagent is
transferred to an analyzer device comprising a porous carrier. The result can
be read from a detection zone(s) of the analyzer device, comprising at least
one specific binding reagent, directly visually or by appropriate equipment
capable of recording the results. Additionally the invention provides an
immunochemical process for determining the presence or absence of an analyte
in a sample solution and further a kit comprising the filter device.


French Abstract

L'invention fournit un dispositif de filtre immunochimique et l'utilisation de celui-ci, ledit dispositif de filtre comprenant un matériau filtre attaché à un élément support, par exemple un bouchon d'un récipient. Le matériau filtre comprend un réactif liant spécifique marqué, ledit réactif liant spécifique marqué étant activé par la filtration d'une solution d'échantillon liquide sur le matériau filtre. On transfère le mélange de la solution d'échantillon et du réactif liant spécifique marqué vers un dispositif analyseur comprenant un support poreux. On peut lire le résultat à partir d'une ou plusieurs zones de détection du dispositif analyseur, comprenant au moins un réactif liant spécifique, directement visuellement ou par un matériel approprié capable d'enregistrer les résultats. En plus l'invention fournit un procédé immunochimique servant à déterminer la présence ou l'absence d'un analyte dans une solution d'échantillon et en plus un kit comprenant le dispositif de filtre.

Claims

Note: Claims are shown in the official language in which they were submitted.


17
Claims
1. An immunochemical filter detection system comprising a test cassette, a
cylindrical cap having a body, a closed end and an open end consisting of a
filter
material consisting essentially of a labeled binding reagent for analyte bound
thereto
in dry form, and a cylindrical sample collection vessel for receiving the
filter material
having an open end that is liquid tightly attachable to the open end of the
cap and an
opposing closed end, wherein the filter material protrudes outwardly from the
open
end of the cap and, is in a path for flow of sample thereinto when the cap is
liquid
tightly attached to the sample collection vessel, and has a particle retention
of 10 to
20 micrometers, wherein the binding reagent is selected from the group
consisting of
an antibody, a recombinant antibody, an antigen, a lectin, a receptor, a
ligand,
fragments thereof and combinations thereof, and wherein further said labeled
binding reagent and any analyte bound thereto are retained within said filter
material
until the cap is removed from the sample collection vessel, applied to an
aperture in
the test cassette and exposed to a binding reagent-free liquid analyte sample
solution for migration through the filter material.
2. The immunochemical filter device according to claim 1 wherein said
sample
solution comprises a liquid sample mixed with a buffer solution.
3. The immunochemical filter device according to claim 1 wherein said
filter
material is selected from the group consisting of polyethylene, polyester,
glass fiber
and composites thereof.
4. The immunochemical filter device according to claim 1 wherein the label
is
selected from colored latex, gold, metal, dye, fluorogenic substances,
superparamagnetic substances, chromogenic substances, fluorochromogens and
enzymatic labels.

Description

Note: Descriptions are shown in the official language in which they were submitted.


CA 02570383 2006-12-14
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Filter device, the method, kit and use thereof
FIELD OF THE INVENTION
The present invention relates to an immunochemical filter device, comprising a
filter
material attached to a support member, e.g. a cap of a vessel. The filter
material comprises
a labeled specific binding reagent, which is activated by filtration of a
liquid sample
solution through the filter material. The mixture of the sample solution and
the labeled
specific binding reagent is transferred to an analyzer device. The invention
further relates to
use of such a filter device and to a process for determining the presence or
absence of an
analyte in a sample with a filter device of the invention. A kit comprising
the filter device is
also provided. The devices of the invention are especially suitable for home
use, use at
doctor's offices and/or by technically untrained staff, involving a minimal
level of skills
from the users.
BACKGROUND OF THE INVENTION
Methods and devices based on immunodiffusion are known from for example US
4,757,002, US 3,990,852 and US 4,562,147. Immunochromatographic methods based
on
lateral flow are known from EP 0 291 194, EP 0 284 232, EP 0 250 137, and WO
86/03839. A diagnostic device comprising a post-filter unit is known from US
application
2003/0049857.
US 4,562,147 provides a radial immunodiffusion enzyme assay method for testing
of
pseudorabies antibodies in swine and other animals. Agar test plates are
provided including
an underlying adherent coating of solubilized non-infectious swine
pseudorabies antigen.
The result of the test is obtained from the diameters of the resulting colored
zones which
correlate with the titers obtained by the official virus neutralization test.
EP 0 291 194 relates to assays involving specific binding, especially
immunoassays and
devices therefore. The analytical test device comprises a hollow casing,
containing a dry
porous carrier, which communicates indirectly with the exterior of the casing
via a bibulous
sample receiving member. The carrier contains in a first zone a labeled
specific binding
reagent and in a second zone spatially distinct from the first zone an
unlabelled specific
binding reagent for the same analyte. When the test is performed the sample
solution is
contacted with the test device directly which increases the risk of overflow
to the second

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2
zone.
EP 0 284 232 provides a solid phase assay for determining the presence or
absence of
analyte in a liquid sample. A test strip of the invention has a tracer movably
supported on a
first portion and a binder immobilized on a second portion. A disadvantage of
this method
is that the sample solution is contacted with the test during the performance
of the test,
which increases the risk of overflow to the second portion.
EP 0 250 137 describes an immunoassay using colloidal gold for detecting a
ligand in a
sample, where a membrane strip is contacted with a sample and simultaneously
or
successively with a liquid reagent containing a ligand binding partner or
ligand labeled with
colloidal gold. A disadvantage of this method is that an additional liquid
reagent containing
the labeled ligand is needed.
WO 86/03839 illustrates a solid phase diffusion assay where the sample is
first mixed with
a labeled binding substance and then applied to a region of a support with
immobilized
adsorbent molecules and allowed to diffuse therein. The diffusion pattern is
visualized and
measured.
US 2003/0049857 relates to a diagnostic device comprising a test unit and a
post-filter unit.
The post-filter unit comprises a label zone containing a dried indicator
reagent. The
indicator reagent is drawn trough the post filter unit with a separate buffer
and the sample is
added directly to the test unit comprising the reaction zone. This however
makes the test
awkward to use.
It is evident from the description of the background art that a multitude of
different test kits
are available. A wide variety of test kits are available commercially and many
of them are
intended for home use. In spite of their convenient formats, there are many
risks for errors
if they are used in an erroneous manner. This risk is imminent when the test
device is used
for sampling, which may be both impractical and inconvenient. Especially, when
the test
device is used as a sampling device or for collecting the sample there is a
risk that the
sensitive reagents and the structure of the analytical device is destroyed or
disturbed.
Further solid samples or liquid samples which need to be diluted require an
additional step.

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3
The present invention provides an improved detection system for
irnmunochemical tests
involving a filter device which enables a liquid sample with or without
dilution or a diluted
solid sample to be filtered through the device and the sample solution
together with the
labeled specific binding reagent to be transferred and mobilized from the
filter material in a
controlled manner. The filter material of the filter device is not sintered.
Due to the two-part
system the analyzer device is not in direct contact with the liquid sample
solution, thereby
minimizing the possibilities that the reagents in and the structure of the
analyzer device are
destroyed or disturbed by the sampling procedure. The device of the present
invention is
especially advantageous for tests where the sample need to be diluted.
SUMMARY OF THE INVENTION
The present invention provides an immunochemical filter device for performing
an
immunoassay comprising a filter material attached to a support member. The
filter material
comprises a labeled binding reagent, preferably a labeled specific binding
reagent which is
activated by filtration of a liquid sample solution through the filter
material, i.e. the labeled
binding reagent is released from the filter material by migration of a liquid
sample solution
therethrough.
The support member is preferably a cap-like member that attaches in a liquid
tight manner
to a sample collection device, such as a tube. The labeled binding
reagent/liquid sample
solution is expressed from the cap (e.g., through an aperture, diffusible
membrane or valve
therein) for application to an analytical device (e.g., an
immunochromatographic test strip
having a binding reagent disposed thereon).
In a preferred embodiment of the present invention the filter device is part
of a cap which is
liquid tightly attachable to a vessel. The liquid sample solution from the
vessel alongside
with the labeled specific binding reagent (preferably a reagent specific for
analyte) released
by contact with the sample from the filter material are transferred from the
filter device to

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the analyzer device comprising lateral flow testing from which the positive or
negative
results are directly readable. The result can be read directly visually (by
naked eye) or by
appropriate equipment capable of recording the results. The sample solution
may contain
the sample as such or diluted with a buffer solution or extraction buffer.
An immunochemical process of the present invention for determining the
presence or
absence of an analyte in a sample solution is also provided. The process
comprises the steps
of adding a sample to a vessel, attaching a filter device comprising a filter
material attached
to a support member liquid tightly to the vessel wherein the filter material
comprises a
labeled binding reagent, preferably a labeled specific binding reagent,
filtering the sample
through the filter material, contacting the mixture of the sample and the
labeled specific
binding reagent with an analyzer device and reading the result by detecting
the presence or
absence of said analyte in the analyzer device. When the sample is caused to
migrate
through the filter material the labeled binding reagent is released.
The present invention further relates to a kit for determining the presence or
absence of an
analyte in a sample. The kit comprises the test device of the invention and a
vessel (e.g., a
sample collection vessel) whereto the filter device is liquid tightly
attachable. The filter
device comprises a filter material attached to a support member (e.g. a cap)
wherein said
filter material comprises a labeled binding reagent, preferably a labeled
specific binding
reagent. Further an analyzer device comprising a porous carrier wherein said
porous carrier
lacks a labeled specific binding reagent may be part of the kit.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a side view of a filter device and a vessel in accordance with the
invention.
FIG. 2 is a side view of a filter device and a vessel in accordance with the
invention, where
the vessel comprises a buffer solution.
FIG. 3 is a view seen from above of an analyzer device.
FIG. 4 is a view seen from above of another embodiment of the analyzer device.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to an immunochemical filter device for
performing an
immunoassay wherein the filter device comprises a filter material attached to
a support
member. The filter material comprises a labeled specific binding reagent which
is activated

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by filtration of a liquid sample solution through the filter material. The
labeled binding
reagent, preferably one specific for analyte, is released from the filter
material by migration
of the liquid sample through the filter material.
In a preferred embodiment the support member is a cap of a vessel, for example
a tube
(e.g., a sample collection tube), and said cap is liquid tightly attachable to
the tube. Said cap
preferably includes an aperture, diffusible membrane or valve through which
liquid sample
solution, after contact with labeled binding reagent, may be caused to flow
from the filter
device.
The invention further provides a detection system comprising an immunochemical
filter
device and an analyzer device, where the mixture of sample solution and
labeled specific
binding reagent or the reaction product (complex) formed thereof are added to
the porous
carrier of the analyzer device comprising a specific binding reagent
optionally immobilized
on the porous carrier. Normally the liquid pressed from the filter device
moves throughout
the porous carrier of the analyzer device by diffusion and/or capillary
action. The liquid
may be expressed from the filter device through an aperture, a diffusible
membrane or a
valve of the cap.
The present invention also relates to a kit for determining the presence or
absence of an
analyte in a sample, comprising a filter device comprising a filter material
attached to a
support member and a vessel whereto the filter device is liquid tightly
attachable. The filter
material comprises a labeled specific binding reagent.
The immunochemical filter device enables a controlled mobilization and mixing
of the
labeled specific binding reagent with a potential analyte of the sample
solution in the
analyzer device comprising a porous carrier. In other words the labeled
specific binding
reagent is mobilized in a controlled manner when transferred to the analyzer
device
comprising a porous carrier. Thereby a more sensitive method for detecting
different
analytes in samples is provided. Furthermore, since the filter device also
filters the sample,
a sensitive and hygienic test is provided. Solid particles of a faeces sample
or another solid
sample will not penetrate or permeate the filter and will thus not block the
pores of the
analyzer device. If a vessel is used, the solid particles stay in the vessel
thus providing a
hygienic test.

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6
The analytes to be detected can be disease specific antibodies including IgG,
IgM and IgA,
antibodies against Helicobacter pylon, Enterohaemorrhagic Escherichia coli
(EHEC),
Myoglobin, Troponin, Toxoplasma, Chlamydia trachomatis and pneumonia,
Bordetella
pertussis, Mycoplasma pneumoniae, Hepatitis A, B, C, 11W1,2, respiratory
disorders or etc.,
autoantibodies against human proteins, viruses from faeces including Rota,
Adeno, Parvo,
Astro or Distemper, antigen tests from food or environmental samples including

Enterohaemorrhagic Escherichia coli (EHEC), antigens excreted to serum
including
Myoglobin and Troponin, antigens excreted to urine including Luteinizing
hormone (LH),
Follicle stimulating hormone (FSH) and human chorionic gonadotropin (hCG) or
to faeces
including H. pylori, samples taken with a swab such as Group A Streptococci,
Chlamydia
antigen tests, Candida, Trichomonas or other antigens of or antibodies against
bacteria,
virus, fungi and parasites or components and products thereof.
The device and method of the invention may be used for a multitude of
different tests
including pregnancy, menopause, fertility, C-reactive protein (CRP), Thyroid
stimulating
hormone, toxoplasmosis, cancer antigen, respiratory disorder, bacteria,
viruses, venereal
diseases, celiac disease, allergy, myocardial infarct and drug tests, etc.
In preferred embodiments of the invention the kit may further comprise a
buffer solution
wherein said buffer solution is added to the vessel in order to make the test
kit ready to use
or an analyzer device comprising a porous carrier wherein said porous carrier
lacks a
labeled specific binding reagent. Further positive or negative controls, a
sampling device,
for example a capillary, pipette or swab may be included. In order to provide
everything
needed to perform a certain test also lancets, alcohol pads and plasters may
be included in
the kit.
The sample can be urine, faeces, blood, serum, plasma, saliva, mucus,
excretion from eyes
or the sample solution may be a mixture of a sample and a buffer solution.
The present invention is advantageous for tests where the sample have to be
diluted in order
to perform a test. This does not require an additional step when the present
invention is
used since the sample may be added to a vessel already containing a suitable
buffer
solution. Further the sample is easy and hygienic to handle and the risk of
contamination is
decreased. Solid samples for example faeces are one example of samples which
need to be

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7
diluted. Furthermore for example many whole blood samples such as finger tip
samples (10
usually have to be diluted 1/50 to 1/200 in order to be able to perform
certain tests. This
applies for instance to CRP-tests where the sample has to be diluted with a
buffer solution
in order to obtain a suitable sample solution.
The buffer of the buffer solution is chosen from buffers known in the art and
depends on
the detected analyte. The pH is usually between 7 and 8,5 and the capacity of
the buffer is
sufficient to maintain the pH. The buffer solution may further comprise
additives in order to
stabilize or increase or decrease other features of the buffer solution. The
buffer solution
may be added to the vessel of the kit beforehand and can thus be included in a
kit of the
present invention.
The filter device of the present invention is especially advantageous for
determining for
example Helicobacter pylori. The filter device is a filter tip comprising an
antibody against
H. pylon or an H. pylon antigen dried as a part of a conjugate solution on the
filter
material. If H. pylon antibodies are detected from whole blood or serum
samples an H.
pylon antigen is used and if the sample is faeces an antibody against H. pylon
is added to
the filter tip.
Another advantageous use of the test device of the present invention is for
determining
celiac disease wherein the filter device in the form of a filter tip comprises
a tissue protein
or anti human IgA specific antibody against transglutaminase.
The filter device is dried to a moisture content of 8 % or less and packed
hermetically
separately or in combination with said analyzer device.
Constructing the filter device
The construction of the filter device comprises the steps of; treating a
filter material
attached to a support member with a conjugate solution comprising a labeled
specific
binding reagent and optionally additives, and further drying, and packing the
device. The
support member is provided with an aperture, diffusible membrane and/or valve
(preferably
one openable on application of gentle pressure; i.e., on pressure of the cap
against the
sample well of an analytical immunochemical device).

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8
In a preferred embodiment the filter material comprising the labeled specific
binding
reagent in one end is attached to a transparent cap, in which an aperture is
disposed. The
filter device is attachable at an end opposing the aperture to a sample
collection vessel.
Different filter materials known in the art are possible to use. Preferably a
hydrophilic
permeable material such as polyethylene, polyester, glass fiber and mixes
thereof is
selected. The particle retention of the filter material is preferably 1 to 25
micrometer, more
preferably 10 to 20 micrometer.
Adding the specific labeled reagent
The filter material of the filter device is impregnated with a conjugate
solution comprising a
specific labeled reagent. The whole material may be impregnated but preferably
the
conjugate solution is added in the middle of one side of the filter material
and impregnated
as a thin layer on top of the filter material. The preferred amount of
conjugate solution is 1
to 100 ill, preferably 10 to 40 il. If the filter device comprises a filter
tip or cap which is
liquid tightly attachable to the vessel, the conjugate solution is dispensed
at that end which
is put inside the vessel.
The labeled specific binding reagent is obtained by coating the label
particles with specific
binding reagent(s) using methods well known in the art. The specific reagents
of the filter
device and the analyzer device include antibodies, antibody fragments,
recombinant
antibodies, recombinant antibody fragments, antigens, lectins, receptors
and/or ligands. The
label applicable in the filter device includes colored latex, gold, metal,
dye, fluorogenic
substances, superpara-magnetic particles coated with the specific binders.
Chromogenic
substances, particularly fluorochromogens and enzymatic labels may be used as
markers as
well. It is possible to use a combination of several different labeled
specific binding
reagents if the sample is detected for more than one analyte and the same
specific binding
reagent cannot be used for all.
The labeled specific binding reagent may be added to the filter material as
such or as a
conjugate solution further comprising additives for example natural or
synthetic polymers
such as albumin (BSA, Bovine serum albumin) and casein or PEG (polyethylene
glycol),
PVA (polyvinyl alcohol) and PVP (polyvinyl pyrrolidone), nonionic detergents
such as
TWEEN 20, ITEXA (hexane sulphonic acid), TRITON-X-100, SDS and BRIJ and

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preservation agents such as sugar, for example glucose, sucrose and trehalose
or derivatives
thereof. Additives are added as stabilizers in order to increase stability, in
order to improve
the release of labeled specific binding reagent and as blocking agents in
order to make the
reactive sites of the filter material inert when the test is performed and the
conjugate
solution is activated by a sample solution. The conjugate solution may be
applied by hand
with a pipette or with an automated dispenser.
Drying and packing the device
The filter device is dried in a dry room, with a relative humidity less than
20 % and further
in a dry room with a relative humidity less than 8 %. The filter device is
then packed
hermetically separately or in combination with an analyzer device, for example
in a
hermetic pouch.
Preparing the analyzer device
The mixture of a sample solution and a labeled specific binding reagent may be
transferred
to an analyzer device, which is the result recording detecting device of the
detection
system. The analyzer device may be prepared by immobilizing one or more
specific
binding reagents and optionally also control reagents directly or indirectly
to a porous
carrier of the device. The porous carrier is thereafter blocked. The blocking
solution, a
mixture comprising natural or synthetic polymers such as albumin (BSA, Bovine
serum
albumin) and casein or PEG (polyethylene glycol), PVA (polyvinyl alcohol) and
PVP
(polyvinyl pyrrolidone), nonionic detergents such as TWEEN 20, ITEXA (hexane
sulphonic
acid), TRITON-X-100, SDS and BRIJ and preservation agents such as sugar, for
example
glucose, sucrose and trehalose or derivatives thereof, is prepared in order to
make the
reactive sites of the porous carrier inert.
The porous carrier of the analyzer device is preferably selected from a group
of materials
consisting of nitrocellulose, paper, glass fiber, nylon, polyester,
polysulphonate or cellulose
and derivatives thereof and the porous carrier can optionally be placed on a
backing and/or
in a casing. The analyzer device of the present invention differs from the
conventional prior
art devices in that it lacks the mobilizable labeled specific binding reagent.
It is obvious for one skilled in the art that the filter device can be used in
connection with a
analyzer device where the porous carrier comprises one porous passage, which
may be

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penetrated by a sample solution, containing detection zone(s) but also with an
analyzer
device where the porous carrier comprises two or more channels optionally made
by a
suitable method comprising at least one specific binding reagent per channel,
immobilized
as a dot or zone. A multiple channel device can be prepared by treating a
porous material
with a suitable method in order to get different channels for tests of
different analytes (or
controls). Thus several analytes can be detected from the same sample by one
single test.
Furthermore several detection zones per channel may be added in order to
perform a semi-
quantitative test. The analyzer device may further comprise for example a
sample pad
comprising polyester or glass fiber.
Different types of analyzer devices are described in background art. Devices
with multiple
channels enable testing of several analytes simultaneously. Markers specific
for different
analytes can be grouped together to form different products. The multiple
channel device
comprises a porous carrier processed by a water-repellency treatment or by
laser in order to
cause a network of channels where the tested sample can migrate. Different
specific
binding reagents may be bound in each channel. Multiple channel analyzer
devices have
been described for example in W02004/086042 Al.
In another preferred embodiment of the invention a multiple channel analyzer
device is
used in order to detect for several analytes from the same sample at the same
time. Every
channel of such device can thereto have several detection zones and/or control
zones.
The structure of the filter device and the analyzer device
FIG. 1 represents a typical filter device and vessel. A filter material 2
comprising a labeled
specific binding reagent 3 in one end is attached to a transparent cap I. The
filter device is
attachable to a vessel 4.
FIG. 2 represents a typical filter device and vessel. A filter material 2
comprising a labeled
specific binding reagent 3 in one end is attached to a transparent cap I. The
filter device is
attachable to a vessel 4 containing a buffer solution 5.
FIG. 3 shows a typical analyzer device seen from above. A casing 6 comprises a
sample
well 7 where the mixture of sample solution and conjugate solution is
transferred. The
detection zone of the porous carrier is placed in the test window 8 and in
case of a positive

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ii
result a visible reaction can be seen there. The control zone of the porous
carrier is placed
in the control window 9 where a visible reaction should be seen whenever the
test is
performed.
FIG. 4 shows another embodiment of an analyzer device. Eight channels 10 are
made by
laser-etching a figure on a porous carrier such as nitrocellulose film.
Treated areas 11 where
markings concerning the tests and the manufacturer may be added during the
laser
treatment separate the channels. Each channel 10 comprises a specific binder
in a detection
zone or dot 12. Also a place 13 (sample receiver) where the mixture of sample
solution and
conjugate solution should be transferred is shown.
EXAMPLE I Respiratory disorder
Example I relates to detection of Streptococcus pyogenes from a throat swab.
A solution of a labeled specific binding reagent is prepared by reacting a
gold solution and
Streptococcus pyogenes specific antibodies. Further a conjugate solution
comprising gold
labeled antibodies in 0,01-0,5 M glysine buffer pH 7,5 to 8,5 with BSA (0,1-
1,0 %), Tween
20 (0,01-0,05 %) and trehalose (0,5-1,5 %) is prepared and 20 tl of the
conjugate solution
is added to a POREX SQ-EASYTm filter tip comprising a high density
polyethylene filter
material with a particle retention of 15 pm. The solution is added to the
filter material of the
filter tip (the filter device of Fig. 1) using a pipette. The filter tip is
then left to dry in a dry
room, with a relative humidity less than 20%. The drying is continued in a dry
room with a
humidity of less than 8 %.

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The analyzer device of Fig. 3 is constructed by immobilizing a Streptococcus
pyogenes
specific antibody on the porous carrier to form a detection zone. A monoclonal
antibody
against the labeled specific binding reagent is immobilized on the carrier to
form a control
zone. The nitrocellulose is then blocked with a blocking solution comprising
BSA (0,1-
5,0%), TRITON-X-100, BRIJ and saccharose. The material is allowed to dry. The
porous
carrier is placed in a casing.
A buffer solution (extract) is prepared by adding 4 drops of extract solution
A; 1-3 M
NaNO3 and four drops of a extract solution B; 0,9 % NaC1 pH 1-3 to a tube just
before the
sample is taken. Then the sample is taken from the throat using a swab and the
swab is
allowed to stand in the tube for two minutes after mixing. Then the swab is
removed and
the filter tip is attached to the tube.
Three drops (about 100-120 ill) of sample solution is pressed through the
filter material and
expressed therefrom into the sample well of the analyzer device. The complex
formed of
the sample and labeled specific binding reagent is allowed to migrate for 5
minutes after
which the result is read in the test window. If red lines are visible both in
the detection zone
of the test window and in the control zone of the control window, it indicates
the presence
of Streptococcus pyogenes in the sample.
EXAMPLE 2 Multiple channel test for C-reactive protein
Example 2 describes use of the invention for C-reactive protein (CRP) and
Myoglobin
testing. The sample is tested against CRP and Myoglobin using a multiple
channel analyzer
device. The test for CRP is in one channel, the Myoglobin test in another and
a control in a
third channel.
A solution of labeled specific binding reagents is prepared by reacting a gold
solution with
C-reactive protein (CRP) specific antibodies and with Myoglobin specific
antibodies. A
conjugate solution comprising gold labeled antibodies in 0,01-0,5 M glysine
buffer pH 7,5
to 8,5 with BSA (0,1-1,0 %), Tween 20 (0,01-0,05 %) and trehalose (0,5-1,5 %)
is prepared
and 40 ill is added to a high density polyethylene POREX SQ-EASYTm filter tip
(filter
device of Fig. 2) and the filter tip is then left to dry in a dry room, with a
relative humidity
less than 20%.

CA 02570383 2006-12-14
WO 2005/124347 PCT/F12005/050215
13
The analyzer device of with three separated channels is constructed by laser-
etching the
channels in a porous carrier of nitrocellulose and a mylard film. 0,5 ill of
two different
specific antibodies are immobilized on the porous carrier to form the
detection zones in two
channels of the analyzer device. An anti-mouse antibody for the labeled
specific binding
reagents is immobilized on the carrier in the third channel to form a control
zone. The
material is allowed to dry before the nitrocellulose is blocked with a
blocking solution
comprising BSA, HEXA and trehalose. The porous carrier is dried.
The amount of analyte to give a positive result is adjusted to 3 il1/m1 for
CRP and to 60
ng/ml for Myoglobin.
The test is performed by adding 200 ill of a modified PBS buffer, 1-5 mM EDTA
and 100
ill of whole blood to a tube. The filter device is attached to the tube and
three drops of the
mixture of sample solution and conjugate solution is pressed trough the filter
material of the
filter tip to that place of the analyzer device were the sample should be
placed. The sample
and the labeled specific binding reagent are allowed to migrate for 5 minutes
after which
the result is read as visible dots in the control zone and in the detection
zones, for samples
where CRP and Myoglobin are present in amounts of 3 ill/m1 and 60 ng/ml or
more.
EXAMPLE 3 Multiple channel test for virus antigen detection
Example 3 describes use of the present invention for detection of virus
antigens of
Rotavirus and Adeno viruses.
A solution of two different labeled specific binding reagents was prepared by
reacting a
latex solution with specific antigens for Rotavirus and Adenovirus. 40 ill of
a conjugate
solution comprising the latex labeled antigens in 0,01-0,5 M glysine buffer pH
7,5 to 8,5
with BSA (0,1-1,0 %), Tween 20 (0,01-0,05 %) and trehalose (0,5-1,5 %) was
prepared and
added to a high density polyethylene POREX SQ-EASYTm filter tip. The filter
tip was then
left to dry in a dry room, with a relative humidity less than 20%. The drying
is continued in
a dry room with a humidity of less than 8 %.
The analyzer device with three separated channels was constructed by forming
the channels
in a porous carrier of nitrocellulose and a mylard film 0,5 ill of two
different antibodies

CA 02570383 2006-12-14
WO 2005/124347 PCT/F12005/050215
14
were immobilized on the porous carrier to form the detection zones in two
channels of the
analyzer device. An anti-mouse antibody for the labeled specific binding
reagents was
immobilized on the carrier in the third channel to form a control zone. The
reactive points
of the channels were blocked and the analyzer device was dried.
100 mg of a faeces sample was added to a tube comprising 900 ill of a PBS pH
7,7 buffer
solution. The sample and buffer solution was mixed and the filter tip was
attached to the
tube and the diluted sample and the conjugate solution of the filter device
was pressed
through the filter material and applied to the sample application spot of the
analyzer device,
from where the mixture diffused and migrated to the test zones, where a
reaction took place
if the sample contained the analyzable virus antigen and a visible dot or line
could be seen.
EXAMPLE 4 Detection of Escherichia coli 0157 and 0111 in food
Example 4 relates to a test for Enterohaemorrhagic Escherichia coli (EHEC).
The labeled specific binding reagent was prepared by reacting a gold solution
and an E. coli
0157 antibody. The conjugate solution and the filter tip were prepared
according to
example 1. An analyzer device comprising an E. coli 0157 specific antibody in
the
detection zone was also prepared according to example 1. The filter tip and
the analyzer
device were hermetically dried and packed in a pouch together with a pipette.
25 mg of solid samples of meat and vegetables were homogenized in a Stomacher
for 2
minutes in 225 ml of modified Trypticase-Soy Broth + Novobiocin and incubated
on a
rotary shaker (100 RPM) at 37 C to 42 C for 7 hours.
Pouches containing the devices were opened and 10 drops of the enriched
samples were
dispensed in tubes using a pipette. The filter tips were placed on the tubes.
Three drops
(approximately 110 of sample solutions together with the conjugate of the
filter devices
were dispensed through the filter material into the sample window of the
analyzer devices.
The results were read 5 minutes after the application of the solution to the
analyzer devices.
Red lines were visible both in the detection zone of the test window and in
the control zone
of the control window, which indicated the presence of E. coli 0157 in the
sample.

CA 02570383 2006-12-14
WO 2005/124347 PCT/F12005/050215
Positive and negative control samples were used to check proper performance of
the tests.
10 drops of a positive control solution were added to a filtering tube and the
testing was
performed as described above. After 5 minutes two visible lines were detected
in the
detection window and the control window.
10 drops of a negative control solution were added to a filtering tube and the
testing was
performed as described above. Only one visible line was detected in the
control window.
EXAMPLE 5 Test for celiac disease
Example 5 relates to a test for celiac disease.
The labeled specific binding reagent was prepared by reacting a gold solution
and a
recombinant tissue transglutaminase. The conjugate solution and the filter tip
were prepared
according to example 1. An analyzer device comprising an anti human IgA
specific
antibody in the detection zone was also prepared according to example 1. The
filter tip and
the analyzer device were hermetically dried and packed in a pouch together
with a tube
containing 500 ill PBS buffer solution, a pipette, a lancet, an alcohol pad
and a plaster.
Positive and negative control samples were used. 1 drop (10 ill) of blood was
added to a
filtering tube containing 500 ill of buffer solution. The filter tip was
attached to the tube
and 1 drop of sample solution was transferred through the filter material to
the analyzer
device. After 5 minutes two visible lines were detected. One of the lines was
in the
detection window and the other was in the control window.
Respectively 1 drop of a negative control solution were added to a filtering
tube and the
testing was performed as described above. Only one visible line was detected
in the control
window.
Performed at home the alcohol pad is used to wash a finger tip from which a
blood sample
is taken using the lancet. 1 drop of blood is transferred to the tube with the
pipette. The test
is performed as described above.

CA 02570383 2006-12-14
WO 2005/124347 PCT/F12005/050215
16
Based on the above description it is evident that use of filter devices of the
present
invention increases security of the test results due to the more controlled
mobilization of the
labeled specific binding reagent. Another advantage of the filter device is
that tests can
easily be performed for samples which need dilution and/or filtration. The
samples may be
added to a vessel already containing a buffer solution or an extraction
buffer.
Further as the test system is easy to use it enables home use. Due to the fact
that the
analyzer device is not in direct contact with the liquid sample solution,
overflow is avoided
and an increased reliability of the test is obtained. Moreover the filter
device and analyzer
device of the invention are easy to store due to the fact that the devices are
dried and that
they are possible to store hermetically.

Representative Drawing
A single figure which represents the drawing illustrating the invention.
Administrative Status

For a clearer understanding of the status of the application/patent presented on this page, the site Disclaimer , as well as the definitions for Patent , Administrative Status , Maintenance Fee  and Payment History  should be consulted.

Administrative Status

Title Date
Forecasted Issue Date 2015-04-14
(86) PCT Filing Date 2005-06-15
(87) PCT Publication Date 2005-12-29
(85) National Entry 2006-12-14
Examination Requested 2010-06-11
(45) Issued 2015-04-14
Deemed Expired 2018-06-15

Abandonment History

Abandonment Date Reason Reinstatement Date
2013-06-28 R30(2) - Failure to Respond 2014-06-23
2014-06-16 FAILURE TO PAY APPLICATION MAINTENANCE FEE 2014-07-08

Payment History

Fee Type Anniversary Year Due Date Amount Paid Paid Date
Application Fee $400.00 2006-12-14
Maintenance Fee - Application - New Act 2 2007-06-15 $100.00 2007-06-13
Registration of a document - section 124 $100.00 2008-04-28
Maintenance Fee - Application - New Act 3 2008-06-16 $100.00 2008-06-06
Maintenance Fee - Application - New Act 4 2009-06-15 $100.00 2009-06-10
Request for Examination $800.00 2010-06-11
Maintenance Fee - Application - New Act 5 2010-06-15 $200.00 2010-06-11
Maintenance Fee - Application - New Act 6 2011-06-15 $200.00 2011-06-15
Maintenance Fee - Application - New Act 7 2012-06-15 $200.00 2012-05-15
Maintenance Fee - Application - New Act 8 2013-06-17 $200.00 2013-05-15
Reinstatement - failure to respond to examiners report $200.00 2014-06-23
Reinstatement: Failure to Pay Application Maintenance Fees $200.00 2014-07-08
Maintenance Fee - Application - New Act 9 2014-06-16 $200.00 2014-07-08
Final Fee $300.00 2015-01-23
Maintenance Fee - Patent - New Act 10 2015-06-15 $250.00 2015-06-11
Maintenance Fee - Patent - New Act 11 2016-06-15 $250.00 2016-05-31
Owners on Record

Note: Records showing the ownership history in alphabetical order.

Current Owners on Record
ANI BIOTECH OY
Past Owners on Record
NISKANEN, AIMO
SARAMAKI, MIKA
Past Owners that do not appear in the "Owners on Record" listing will appear in other documentation within the application.
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Document
Description 
Date
(yyyy-mm-dd) 
Number of pages   Size of Image (KB) 
Abstract 2006-12-14 2 90
Claims 2006-12-14 4 123
Description 2006-12-14 16 819
Drawings 2006-12-14 4 1,000
Representative Drawing 2006-12-14 1 33
Cover Page 2007-02-15 1 48
Claims 2006-12-15 4 118
Description 2012-09-06 16 790
Claims 2012-09-06 5 156
Description 2012-09-07 16 807
Claims 2012-09-07 5 161
Claims 2014-06-23 1 41
Representative Drawing 2015-03-12 1 180
Cover Page 2015-03-12 1 156
Assignment 2006-12-14 3 78
PCT 2006-12-14 3 112
Prosecution-Amendment 2006-12-14 5 147
Correspondence 2007-02-13 1 26
Fees 2007-06-13 1 39
Correspondence 2008-02-22 2 35
Assignment 2008-04-28 2 59
Fees 2009-06-10 1 201
Assignment 2009-05-21 2 51
Fees 2010-06-11 1 201
Prosecution-Amendment 2010-06-11 1 41
Prosecution-Amendment 2012-03-06 3 105
Prosecution-Amendment 2012-12-28 2 64
Prosecution-Amendment 2012-09-06 12 454
Prosecution-Amendment 2012-09-07 13 501
Prosecution-Amendment 2014-06-23 3 139
Fees 2014-07-08 1 33
Correspondence 2015-01-23 1 45