COMPOSITIONS FOR FABRIC BASED LATERAL FLOW ASSAY DEVICE USING ELECTROCHEMICAL DETECTION MEANS  AND DEVICES THEREFROM
TECHNICAL FIELD
[0001] The invention relates generally to fabric based woven lateral flow assay devices comprising electrochemical detection means.
BACKGROUND
[0002] Lateral flow assay devices have been found to be a very convenient choice of device for many applications  especially in point-of-care diagnostics. Several different types of lateral flow assay devices are known in the art. Most lateral flow assay devices are made from a paper based or nitrocellulose based material to construct the device. More recently  fabric based lateral flow assay devices have garnered greater attention. For example  WO2009/121043 describes a cotton based lateral flow assay device. Reches et al (Reches et al  Applied Materials and Interfaces (2010)  Vol. 2 No. 6  pp. 1722-1728) describe lateral flow assay devices based on cotton. In that report  the authors have attempted using the following materials as choice for lateral flow assay devices: nitrocellulose  cotton  polyester  nylon  natural silk  hemp  wool  acrylic fiber  a 50/50 blend of acrylic and cotton  and have categorically stated that the possible choices of material for a lateral flow assay device would be cotton  nylon and polyester  with cotton being the best and only choice for further studies.
[0003] WO2012/004635 and WO2012/004636 however describe the successful use of natural silk and appropriately treated silk as starting material for lateral flow assay devices. This opens up the possibility for woven silk fibers to be used for such practical applications. Silk comprises a polypeptide chain obtained from natural sources and has been found to be biocompatible  and further comprises functional groups for any further modifications.
[0004] There is a need to develop the silk based lateral flow assay devices with a view to render it useful in a variety of test conditions for several different diagnostics testing situations.
BRIEF DESCRIPTION
[0005] In one aspect  the invention provides a diagnostic comprising at least one fabric strand coated with a conducting material and a diagnostic material.
[0006] In another aspect  the invention provides a lateral flow assay device having a proximal end  a distal end and a testing zone intermediate between the proximal end and the distal end  wherein the lateral flow assay device is made of a fabric comprising at least one strand to define a flow path  wherein the at least one strand of the fabric in the testing zone is coated with a conducting material  wherein the at least one strand is made of a silk-based material.
[0007] In yet another aspect  the invention provides a lateral flow assay system comprising the lateral flow assay device of the invention.
DRAWINGS
[0008] These and other features  aspects  and advantages of the present invention will become better understood when the following detailed description is read with reference to the accompanying drawings in which like characters represent like parts throughout the drawings  wherein:
[0009] FIG. 1 shows a diagrammatic representation of an exemplary embodiment of the lateral flow assay device of the invention;
[0010] FIG. 2 shows a visual depiction of a plurality of strands woven together to form the lateral flow assay device of the invention;
[0011] FIG. 3 shows an exemplary lateral flow assay device of the invention;
[0012] FIG. 4 shows another exemplary lateral flow assay device;
[0013] FIG. 5 shows an exemplary lateral flow assay system of the invention;
[0014] FIG. 6 shows an image of the woven fabric as embodied by Example;
[0015] FIG. 7 shows a single strip cut from the woven fabric seen in Fig. 6 to form the lateral flow assay device;
[0016] FIG. 8 shows amperometric profiles for samples containing different concentrations of glucose in phosphate buffered saline as measured by the lateral flow assay device of the invention;
[0017] FIG. 9 shows a plot of the current output from a control lateral flow assay device for the different concentrations of the aqueous glucose solutions; and
[0018] FIG. 10 shows a plot of peak current against glucose concentration as measured by a control lateral flow assay device.
DETAILED DESCRIPTION
[0019] The definitions provided herein are to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.
[0020] As used in this specification and the appended claims  the singular forms "a"  "an"  and "the" encompass embodiments having plural referents  unless the content clearly dictates otherwise.
[0021] Unless otherwise indicated  all numbers expressing feature sizes  amounts  and physical properties used in the specification and claims are to be understood as being modified in all instances by the term "about." Accordingly  unless indicated to the contrary  the numerical parameters set forth in the foregoing specification and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by those skilled in the art utilizing the teachings disclosed herein.
[0022] As used in this specification and the appended claims  the term "or" is generally employed in its sense including "and/or" unless the content clearly dictates otherwise.
[0023] Strand as used herein refers to a single element (as a yarn or thread) of a woven or plaited material.
[0024] Analyte  as used herein  refers to a substance or chemical constituent that is determined in an analytical procedure. For instance  in an immunoassay  the analyte may be a protein ligand or a binder  while in blood glucose testing  the analyte is glucose. In one instance  the analyte could be a surface antigen that is a marker for the Hepatitis-B virus. In another exemplary instance  analyte may include a drug to be detected  such as cocaine from a blood analysis. The analytical procedure may include  for instance  fluorescence  mass-spectrometry  colorimetry  radio-imaging  electrochemical detection and the like  and combinations thereof. In some instances  analytes may refer to antibodies. In other instances  analytes may refer to antigens. In some other instances  analytes may include small molecule metabolites. Exemplary small molecule metabolites that are tested for diagnostic  therapeutic or other purposes include  but not limited to blood gas  metal and other non-metallic ions  electrolytes  other components like urea  creatinine  creatine etc.  cells like CD-4 CD-8 that are useful as markers for viral load  and the like  and combinations thereof.
[0025] Antibody as used herein refers to protein that is used in the identification of specific antigen. The specific antigen is typically a marker of a disease or certain types of diseases. Sometimes  antibodies may also be referred to as immunoglobulins. Antibody may be primary or secondary antibody. Primary antibodies are antibodies raised against a specific antigen and are generally unlabelled. Primary antibodies may also be referred to as capture antibodies. Secondary antibody is an antibody that binds to primary antibody or fragments contained within the primary or capture antibody. Secondary antibody comprise label that render them useful for detection. Typical labels include fluorescence moiety  radio-active compounds  enzyme-linked labels  magnetically active particles  nanoparticles  quantum dots  latex particle labels  and the like  and combinations thereof. Depending on the label  the method used to detect  identify and quantitate may include fluorescence spectroscopy  radio-imaging  ELISA test and the like.
[0026] Antigen  as used herein  refers to a molecule that is recognized by an immune system of a living organism. Antigen also refers to molecular fragments that may be recognized by the immune system. It is generally known that a given antigen shows specificity to an antibody  and this property of an antigen is used in a variety of applications.
[0027] Silk as used herein refers to a fiber obtained from silkworms  more specifically from the larvae of mulberry silkworms. The silk most useful in the invention are those that can be woven into textiles  such as that obtained from the silkworm Bombyx Mori  however  other forms of silk that may be synthetically made or produced from other sources may also be used for this invention. Chemically  silk fiber comprises a chain of amino acids  which possesses functional groups. These functional groups may be used for binding useful moieties. As used herein  functional groups are reactive chemical moieties that can interact with other reactive species to form physical or chemical bonds.
[0028] Cotton as used herein refers to the cellulosic fiber made available from the genus Gossypium. It can be used as such  or may be modified to render some useful properties to the final fabric strand. Techniques and chemicals for the modification of naturally occurring cotton to improve its properties are known in the art. One exemplary process includes mercerization of cotton. Another process involves preparation of cellulose xanthate.
[0029] As noted herein  in one aspect the invention provides a diagnostic composition that comprises at least one fabric strand coated with a conducting material and a diagnostic material. In one embodiment  the at least one fabric strand is made of cotton. In another embodiment  the at least one fabric strand is made of silk. The silk may also be modified to suit the requirements of the device being fashioned. The modifications may include physical and chemical modifications. A particularly useful modification includes subjecting the silk fiber to a process of degumming. Methods and compositions for degumming are known in the art and can be suitably applied in the invention. Subsequently  the silk fiber may further be coated appropriately to render useful properties. Exemplary coatings include  for example  but not limited to  a surfactant  a hydrophilic coating  a hydrophobic coating  and the like  and so on. The choice of coating and the manner of applying the coating depends on various factors such as extent of hydrophilicity/ hydrophobicity  flow characteristics etc. and will become apparent to one skilled in the art.
[0030] The silk-based material may further comprise suitable modifications to enable its efficacy as a diagnostic composition. Thus  in an exemplary embodiment  the at least one strand of silk-based material comprises a suitable antibody for a specific antigen to detect the presence or absence of an analyte in the test sample. Methods of linking such antibodies to the silk based materials are known  and may include chemical and physical modifications.
[0031] Useful conducting materials in the invention include  for example  but not limited to  carbon ink  silver halide  gold  platinum  conducting polymers such as polyaniline  PEDOT/PSS  and the like  and combinations thereof. The conducting material  when it comes in contact with a specific material or types of materials  becomes electrically active giving rise to electric signals. These electric signals are then capable of being detected using appropriate detection means.
[0032] The diagnostic material useful in the invention may be any of the analytes described herein  or any reagent specific to a given analyte that can then be used to test the presence or absence of it  or alternately  measure the amount of the analyte present. The analyte or the specific reagent for the analyte is used to stimulate the conducting material upon contact giving rise to electric signals  which is then measured to provide a suitable output. In one specific embodiment  the diagnostic material is glucose oxidase  which when it comes in contact with glucose effects a redox reaction giving rise to an electron that leads to an electrical signal. The extent of the electrical signal may then be used to quantitate the amount of glucose present using a suitable predetermined calibration curve.
[0033] In another aspect  the invention provides a lateral flow assay device. Fig. 1 shows a diagrammatic representation of an exemplary embodiment of the lateral flow assay device of the invention generally represented by numeral 10. The device of the invention 10 comprises a proximal end depicted by numeral 12 in Fig. 1  a distal end 14 and a testing zone 16 that is present intermediate between the proximal end 12 and the distal end 14. The lateral flow assay device is made of a fabric comprising at least one strand that defines a flow path  shown in Fig. 1 by numeral 18. In one exemplary use case scenario  a sample is introduced onto the lateral flow assay device at the proximal end side of the device. Subsequently  the sample is allowed to flow along the flow path from the proximal end 12 to the distal end 14.
[0034] The at least one strand of the fabric is made of a silk-based material. The silk as described herein may be used as such to form the lateral flow assay device of the invention. Methods of making the lateral flow assay device are described in the art  in for example  WO2012/004635 and WO2012/004636. One particularly useful method for making the device includes weaving a plurality of strands together. Fig. 2 shows a visual depiction of a plurality of strands woven together to form the lateral flow assay device of the invention.
[0035] The lateral flow assay device of the invention is made by the inter-weaving strands of the silk-based fiber. The method of inter-weaving is generally known among textile manufacturers. A useful method of inter-weaving includes the use of a double warp double weft technique. Other methods of introducing warp and weft are known in the art. Warp means the lengthwise yarns while weft means the yarn that is threaded through the warp. The patterning frequency of the warp and weft structure can influence the flow properties within the strands of fiber  and hence provides greater control over the use of the device of the invention. In addition  yarn properties such as ply  twists per inch and surface properties can also be used to tune wicking properties along the length of the fabric. Currently  machines exist that are used extensively in the textile manufacturing industry for the production of finished textile goods using the double warp double weft technique along with other techniques.
[0036] A loom is an exemplary device used for weaving  and may be advantageously used for the production of the diagnostic device of the invention. Several types of looms are readily commercially available for use. Exemplary looms useful in the invention include  but not limited to  jacquard loom  dobby loom  treadle loom  power loom  and the like.
[0037] The inter-weaving may also be effected using strands from different types of fibers. For example  a silk-based fiber may be interwoven with a cotton-based fiber. In another example  silk-based fiber may be interwoven with cotton-based fiber and a polyester-based fiber. Other useful fibers for the invention  include  for example  but not limited to  nylon  polypropylene  viscose  carbon fiber  graphene  and the like  and combinations thereof. In one particular embodiment  the silk-based fiber may be interwoven with a carbon fiber. Carbon fiber provides the advantage of being conducting in itself  and may or may not require any further process steps to render it more conducting. The relative amounts of each fiber used for preparing the final device depends on various factors such as strength  pliability  economics  and the like  and will become obvious to one skilled in the art. Lateral flow assay devices made by interweaving multiple types of fibers are described in  for example  international application PCT/IB2011/052753.
[0038] In the lateral flow assay device of the invention  the at least one strand in the testing zone is coated with a conducting material. The at least one fabric strand used for coating may be silk in one embodiment  and cotton in another embodiment. Useful conducting materials in the invention include  for example  but not limited to  carbon ink  silver halide  and the like  and combinations thereof. The conducting ink  when it comes in contact with a specific material or types of materials  becomes electrically active giving rise to electric signals. These electric signals are then capable of being detected using appropriate detection means. Fig. 3 shows an exemplary lateral flow assay device of the invention represented by numeral 20 made by weaving a plurality of strands 22. In this exemplary depiction  one strand is coated with a conducting material  shown by numeral 24. The at least one strand coated with a conducting material further comprises a reagent used for testing purposes. Such reagents include  for example a glucose oxidase  to enable the device for testing for blood glucose levels. Other materials may also be used to coat the at least one fabric strand that can facilitate electron transfer rendering the yarn electrically active. Such extraneous materials include  for example a mediator. A useful mediator material is potassium ferricyanide. As used herein  working electrode refers to the fabric strand that is coated with the conducting material and diagnostic material; a reference electrode refers to the fabric strand that is coated with the reference conducting material  and counter electrode refers to the fabric strand that is coated with the conducting material. The working electrode may further comprise a mediator in some instances.
[0039] In some embodiments  the strands of the lateral flow assay device of the invention comprise more than one conducting material  wherein one of the conducting materials is used as a reference to calibrate signals arising from the other conducting materials. A useful reference conducting material is silver halide. Fig. 4a shows another exemplary lateral flow assay device 26 made by weaving a plurality of strands that comprises a working electrode and a reference electrode  wherein the working electrode 28 of the lateral flow assay device is made by coating one strand with a conducting material and a diagnostic material while the reference electrode 30 is made by coating another strand with a reference conducting material. Fig. 4b shows yet another exemplary lateral flow assay device 32 made by weaving a plurality of strands that comprises a working electrode  a reference electrode and a counter-electrode  wherein the working electrode 34 is made by coating one strand with a conducting material and a diagnostic material (i.e. enzyme and mediator in one instance  or enzyme alone in another instance)  the reference electrode 36 is made by coating another strand with a reference conducting material and the counter electrode 38 is made by coating yet another strand with a conducting material.
[0040] Methods of coating are known and may include  for example  dip coating. In one embodiment  the fabric strand is first coated with the conducting material and then coated strand is used for weaving to make the lateral flow assay device. Alternately  the conducting material is printed onto the woven strands to make the lateral flow assay device. Other techniques will become obvious to one skilled in the art and is contemplated to be within the scope of the invention.
[0041] The coating is done such that the final fabric strand is rendered predefined properties. Useful properties that define the fabric include for example  resistance  tensile strength and so on  and combinations thereof. In one embodiment  the resistance of the working electrode is maintained less than about 1000 ohms/cm. In other embodiments  the resistance of the counter electrode is maintained less than about 1000 ohms/cm. In further embodiments  the fabric is coated with a diagnostic material is present at a suitable concentration that allows for measurable output. For example  when the diagnostic material is an enzyme such as glucose oxidase  it may be at least about 1 Unit per active length. Active length as used herein refers to the length of the testing zone of the lateral flow assay device.
[0042] In further embodiments  the strands of the lateral flow assay device may also be coated with other conducting material without any of the reagents  antibodies  antigens  or the like. In such instances  such strands may also be referred to as counter electrodes. Counter electrode when present generally includes only the strand coated with conducting ink and is generally used as a reference point for maintaining a constant potential to improve accuracy of readings.
[0043] The lateral flow assay device of the invention may further comprise colorants  emollients  other additives for various purposes  along with those mentioned herein. These additives may be for cosmetic purposes  to provide extra features  or add greater functionality to the existing lateral flow assay device. Further  the lateral flow assay device of the invention may be mounted onto a substrate. The substrate may be present to provide strength and mechanical integrity to the device. The substrate may be chosen from any number of strong materials known to those skilled in the art  and may include  for example  metal backing such as steel  iron  titanium  alloys  and the like  plastics such as poly(methyl methacrylate)  polystyrene  polyethylene  polypropylene  and the like  cardboard  wood  and others  and combinations thereof.
[0044] The lateral flow assay device of the invention may further be encased in a suitable enclosure to protect it from environmental factors  such as handling during transportation  sunlight  moisture  humidity  and so on. In such instances  the enclosure may be designed in such a way that it can be opened to allow access to the device. Alternately  the enclosure may be present in such a way that there is an opening only for the sample introduction  so that the rest of the device is totally enclosed even during operation. Enclosures suitable for the device may have propertises such as transparency  strength  water resistance  moldability  and the like. Some useful materials that can perform well as enclosures for the device may include  but not limited to  glass  plastics such as poly(methyl methacrylate)  polystyrene  polyethylene  polypropylene  and the like.
[0045] The invention as described herein provides weaving as an alternate manufacturing technology for the manufacture of fabric-based lateral flow assay devices that may also be referred to as `fab chips’. Such devices may be reusable type or may be a single-use  disposable device. Fabric weaving is an art that has developed to a very high degree of skill in many parts of the world  and intricate patterns whose dimensions are limited only to the thickness of an individual thread may be woven in a highly parallelized manner. This technique is capable of being adapted for the manufacture of the devices of the invention in a facile manner. By weaving together strands of fibers along with fibers that are coated with reagents  it is possible to manufacture a very large number of diagnostic devices  which are capable of single-plexing and multiplexing. This will result in the ability to actually fabricate an entire diagnostic device using only a loom. This provides a great advantage over the existing techniques which uses different manufacturing methods for various parts of the diagnostic device  thus complicating the process. Further  unlike the currently commercially available diagnostic devices  such as the paper-based devices which require different materials to function as the conjugation pad  absorption pad and sample delivery pad  the invention provides a single uniform manufacturing process to make the lateral flow assay device.
[0046] During manufacture  a large strip of the lateral flow assay device of the invention may be made and subsequently  smaller strips may be cut  which are then taken for further processing  such as gluing onto a substrate  enclosing in a housing  and the like. The nature of the manufacturing process and device qualities allow for the possibility of patterning complex fluid geometries using standard weaving techniques. Further  an input sample may be split into multiple branches using a combination of hydrophilic and hydrophobic yarn during the weaving step  thus enabling multiplexing of tests. In one exemplary embodiment  the cotton fabric is interwoven with silk such that the cotton thread is present 3 millimeters (3 mm) apart  in another embodiment  the cotton fabric is interwoven with silk such that the cotton thread is present 1 mm apart  and in yet another embodiment  the cotton fabric is interwoven with silk such that the cotton thread is present 5 mm apart. The cotton may then be coated with a combination of a mediator  conducting material and diagnostic material in any suitable order. Alternately  cotton that is already coated with the conducting material  diagnostic material and the mediator is used for the interweaving with silk-based matrix.
[0047] In a use case situation of the lateral flow assay device of the invention  sample that may or may not comprise one or more analyte is introduced into the device at a sample introduction zone. The sample is then allowed to flow through the device thus allowing it to come into contact with the diagnostic material in its path. Analyte  if present  reacts with the diagnostic material on the conducting yarn  causing a chemical reaction that generates an electron transfer. These electrons (sometimes through a mediator) generate an electrical signal. The electron transfer rate is correlated with the concentration of the analyte which can then be measured using known techniques. Without being bound to any theory  it is known that the amount of electrons transferred is proportional to the concentration of analyte. One skilled in the art will also recognize that devices described herein may be used as single-plex devices or multiplex devices  by selecting the appropriate one or more diagnostic materials and the electrode materials as described herein depending on the requirements. The flow of the sample is curbed when there are no more flow regions available. Such flow-stop regions may be engineered using known techniques  and may include  for example absorption pads  increased hydrophobic matrix regions  and so on.
[0048] The device of the invention may be used for any assays to be performed in a wide variety of applications. For example  in case of using the device of the invention for a label-free immunoassay wherein direct detection of analyte can be effected by the appropriate use of a specific binding reagent such as antibody  following which the detection can occur through any one of cyclic voltammetry  chronoamperometry  impedimtery  pulsed amperommetric detection  and the like  and combinations thereof. In another exemplary situation  in case of using the device of the invention for a sandwich immunoassay  sample for qualitative detection of antigen in the sample is introduced on to the sample port of the lateral flow assay device. After introduction  as the sample flows along the predefined path from proximal end to the distal end due to capillary action  it comes in contact with secondary reagent (detection antibody) first in the testing zone  as it is present upstream from the primary reagent (capture antibody). Analyte (antigen) if present  forms a first complex with secondary reagent. Following this  the flow of the solution comprising the first complex reaches the primary reagent (capture antibody) forming a second complex comprising the analyte  primary reagent and secondary reagent. The complex induces a change in the conductivity of the conducting material  which is then read out using appropriate electrical measurement means. In some instances  an electroactive material such as ferrocene may be linked through physical or chemical means to the secondary reagent. The electroactive material will serve to improve the output from the device. If the sample does not comprise the analyte  then the first complex and the second complex does not form  thus no change in the conductivity of the conducting material relative to control conditions is discernible  and the sample flows until it reaches the absorption port. The solution stops flow when there are no more flow regions remaining on the device.
[0049] In another aspect  the invention provides a lateral flow assay system comprising: a lateral flow assay device as described hereon. Fig. 5 shows an exemplary lateral flow assay system of the invention  depicted by the numeral 40. The lateral flow device depicted by numeral 42 comprises a proximal end 44  a distal end 46 and a testing zone 48 intermediate between the proximal end and the distal end. The at least one strand of the fabric is coated with a conducting material in the testing zone  depicted by numeral 50. The strand coated with the conducting material is connected to at least one electrode 52  which in turn is connected to measurement means 54 to measure the output from the at least one electrode. Measured output from the at least one electrode incudes current  voltage  resistance  impedance  and the like  and combinations thereof. The lateral flow device may further comprise a reference material coated onto one of the strands to provide a reference output during test conditions.
EXAMPLE
[0050] A 500 micrometer (?m) cotton thread was obtained from a local commercial source and used as such. Silk thread with 22 Denier monofilament  bivoltine was obtained from Silk Touch  Bangalore  India. Carbon ink and Ag/AgCl was purchased from Creative Materials  USA. Potassium Ferricyanide and Glucose Oxidase having a concentration of 192 U/mg was obtained from Sigma-Aldrich  USA. Water as used in the examples described herein means deionized water.
[0051] Silk threads were converted into 4 ply 25 twist per inch (TPI) and then degummed using the procedure described in publication number WO2012/004635. All the coatings were achieved by dip coating technique. Working electrode was prepared by coating cotton thread with carbon ink and potassium ferricyanide followed by Glucose Oxidase. A range of concentrations of each component was used for testing purposes. Reference electrode was prepared by coating cotton with Ag/ AgCl ink such the concentration of the ink gives rise to a resistance ranging from about 250 -300 Ohm/cm. Counter electrode was prepared by coating carbon ink on the cotton thread. The appropriately coated cotton threads were woven along the weft direction with degummed silk such that the coated cotton was present at a distance of 3 mm intervals to make the working electrode  reference electrode and counter-electrode respectively for each device  wherein the degummed silk was used in the warp and weft direction. Fig. 6 shows an image of the lateral flow assay described here. A single strip about 4 mm wide containing three electrodes was then cut from the woven fabric shown and connections were made from the three electrodes to a potentiostat as shown in Fig. 7.
[0052] Samples of water spiked with no glucose  2.5Mm  10 mM glucose and 15 mM glucose were then used to test the performance of the lateral flow assay devices. The liquid solutions wetted the entire strip through capillary action after being input in the sample entry port and initiated a reaction at the working electrode surface which had been immobilized with enzyme. The redox reaction catalyzed by the Glucose Oxidase and mediated by Potassium Ferricyanide releases electrons to produce a current at the working electrode. A standard amperometric technique which applies a constant potential and then measures the current profile versus time was used. Data from different samples tested were plotted and shown in Fig. 8. The bottom line that remains unchanged over the entire period was obtained from water sample with no glucose  while the other curves are for aqueous glucose solutions having concentrations of 2.5 millimoles (mM)  10 mM and 15 mM respectively. It can be seen that current produced at the electrode is proportional to the concentration of glucose in the sample  while the control phosphate buffered saline having no glucose (PBS) gives a flat readout. This demonstrates that different concentrations of glucose can be detected by analyzing the data. It must be noted that any current value at a given time can be used for measurement and analysis purposes.
[0053] A control lateral flow assay device comprising counter electrode  a reference electrode and another counter electrode was prepared. This device was subjected aqueous glucose solutions of various concentrations and the output current was measured  which is shown in Fig. 9. Fig. 10 shows a graph of the peak current (at time = 0 seconds) versus glucose concentration. It can be seen that the peak output current is concentration independent when the diagnostic material glucose oxidase is absent in the control device.
[0054] A calibration curve was then obtained for the lateral flow assay device of the invention for a series of aqueous solutions comprising known concentrations of glucose. Fig. 11 shows a plot of the peak output current from the device for the different concentrations of the aqueous glucose solutions. It can be seen that a linear relationship in the concentration range shown here exists.
[0055] While only certain features of the invention have been illustrated and described herein  many modifications and changes will occur to those skilled in the art. It is  therefore  to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

We Claim:
1. A diagnostic composition comprising at least one fabric strand coated with a conducting material and at least one diagnostic material.
2. The diagnostic composition of claim 1 wherein the at least one fabric strand is selected from a group comprising cotton  silk  viscose  polypropylene  nylon  or combinations thereof.
3. The diagnostic composition of claim 2 wherein the at least one fabric strand is cotton.
4. The diagnostic composition of claim 3 further comprising silk strand.
5. The diagnostic composition of claim 1 wherein the diagnostic material is glucose oxidase.
6. A lateral flow assay device comprising the diagnostic composition of claim 1.
7. A lateral flow assay device having a proximal end  a distal end and a testing zone intermediate between the proximal end and the distal end  wherein the lateral flow assay device is made of a fabric comprising at least one strand to define a flow path  wherein the at least one strand of the fabric in the testing zone is coated with a conducting material and a diagnostic material  wherein the testing zone is made available in a matrix made of degummed silk.
8. The lateral flow assay device of claim 7 further comprising a sample zone.
9. The lateral flow assay device of claim 7 further comprising a sample reservoir.
10. The lateral flow assay device of claim 7 wherein the at least one strand of the fabric in the testing zone coated with a conducting material is made of cotton.
11. The lateral flow assay device of claim 7 wherein the conducting material is selected from a group comprising carbon ink  gold  platinum  or combinations thereof.
12. The lateral flow assay device of claim 7 wherein the at least one strand of fabric in the testing zone coated with a conducting material is further coated with a mediator to provide a working electrode.
13. The lateral flow assay device of claim 12 wherein the mediator is potassium ferricyanide.
14. The lateral flow assay device of claim 12 wherein the concentration of the conducting material and the mediator gives a resistance of the working electrode less than about 1000 ohms/cm.
15. The lateral flow assay device of claim 12 wherein the diagnostic material is present at a concentration of at least about 1 Unit per active length of the working electrode.
16. The lateral flow assay device of claim 7 further comprising a reference electrode  wherein the reference electrode comprises at least one strand of the fabric in the testing zone is coated with a reference conducting material.
17. The lateral flow assay device of claim 16 wherein the reference conducting material is a silver/ silver halide ink.
18. The lateral flow assay device of claim 7 further comprising a counter electrode  wherein the counter electrode comprises at least one strand of fabric coated with a conducting material such that the concentration of the conducting material gives a resistance of the counter electrode less than about 1000 ohms/cm.
19. The lateral flow assay device of claim 7 further comprising a substrate to support the fabric.
20. The lateral flow assay device of claim 19 wherein the substrate is made of a material selected from a group comprising plastic  metal  wood  cardboard  or combinations thereof.
21. The lateral flow assay device of claim 7 further comprising a housing to house the fabric.
22. The lateral flow assay device of claim 21 wherein the housing comprises at least one of a sample introduction port  a transparent viewing port  or combinations thereof.
23. The lateral flow assay device of claim 21 wherein the housing is made of plastic  metal  wood  cardboard  or combinations thereof.
24. A lateral flow assay system comprising the lateral flow assay device of claim 7.
Rachna Singh Puri
Authorized Representative
Date: 14 August 2012

COMPOSITIONS FOR FABRIC BASED LATERAL FLOW ASSAY DEVICE USING ELECTROCHEMICAL DETECTION MEANS  AND DEVICES THEREFROM
ABSTRACT
[0056] The invention provides a composition for lateral flow assay device that comprises at least one fabric strand coated with a conducting material and a diagnostic material. The invention also provides a lateral flow assay device having a proximal end  a distal end and a testing zone intermediate between the proximal end and the distal end. The device is made of a fabric comprising at least one strand that defines a flow path. At least one strand of the fabric in testing zone is coated with a conducting material. At least one strand is made of at least one of a cotton  a silk-based material  and combinations thereof. The invention also provides a lateral flow assay system comprising lateral flow assay device of the invention  wherein strand coated with conducting material is connected to at least one electrode; and measurement means to measure output from the at least one electrode.