METHODS AND COMPOSITIONS FOR ANALYTE DETECTION

• US 20080199851A1
• Filed: 02/21/2007
• Published: 08/21/2008
• Est. Priority Date: 02/21/2006
• Status: Active Grant

• Current Assignee: Nexus Dx, Inc. (Sphingotec GMBH)
• Original Assignee: Nexus Dx, Inc. (Sphingotec GMBH)
• Inventors: Lidgard, Graham Peter, Johnson, Christopher Johann, Booker, David Dickson, Egan, Richard Laswell
• Granted Patent: US 7,932,099 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0832 cylindrical, tube shaped
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 2400/0683 mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 3/5023 with a sample being transported to, and subsequently stored in an absorbent for analysis
• CPC Class Codes: B01L 3/5029 using swabs
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: G01N 2333/11 Orthomyxoviridae, e.g. influenza virus
• CPC Class Codes: G01N 2469/10 Detection of antigens from microorganism in sample from host
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

The present invention is directed to methods and apparatus for detection of one or more analytes. Analytes include agents or components of infectious agents such as pathogenic virus, as well as enzymes, proteins and biomarkers.

Claims

1. A sample collection device comprising a body comprising:

(a) an upper sealed chamber containing one or more solutions, and one or more breakable seals;

(b) a lower chamber in fluid communication with the upper chamber containing an extraction buffer and/or reagents;

(c) a sample collection implement in fluid communication with said upper scaled chamber; and

(d) wherein said reagents comprise a plurality of specific binding agent (SBA) sandwich pairs, each pair comprising;

(i) an SBA-capture moiety conjugate comprising an SBA which specifically binds a target antigen and a capture moiety;

(ii) an SBA-label conjugate, comprising an SBA which also specifically binds said target antigen, and wherein the pair can form a sandwich with said target antigen; and

wherein the plurality of pairs are directed against different antigens.

2. The sample collection device of claim 1 wherein the capture moiety is all oligonucleotide, avidin, streptavidin, pyranosyl RNA (pRNA), aptamer or a combination thereof.

3. The sample collection device of claim 2 wherein said oligonucleotide is DNA or RNA.

4. The sample collection device of claim 1 wherein the label is a fluorophore, chromophore, metal or a combination thereof.

5. The device of claim 1, wherein said capture moiety is pRNA and said label is Europium.

6. The sample collection device of claim 1 wherein the target antigen is one or more influenza viruses.

7. The sample collection device of claim 1, wherein said influenza virus includes subtypes of a formula HxNy, where x is 1 through 16, and y is 1 through 9, or any combination of xy thereof.

8. The sample collection device of claim 1 wherein said extraction buffer is lyophilized.

9. The sample collection device of claim 1 wherein said reagents are separate or combined together in a pellet, a powder, a pill, a bead, a pressed lyophilized powder, or are dried on a solid support.

10. The sample collection device of claim 1 further comprising a mixing area -containing an adsorbent material located in said lower chamber.

11. The device of claim 1, wherein said upper sealed chamber comprises at least two separate subchambers.

12. The device of claim 11, wherein said subchambers contain solutions that are the same or different.

13. The device of claim 1, which further comprises a membrane that filters particulates from said sample.

14. The device of claim 1, wherein said sample collection implement is a swab.

15. The device of claim 1, wherein said sample collection implement comprises foam, a brush structure, flocculated plastic or spun fiber.

16. The device of claim 14, wherein said swab is upstream of a mixing area.

17. The device of claim 13, wherein said reagents are comprised in said membrane.

18. The device of claim 1, wherein said seal functions as a valve.

19. The device of claim 15, wherein said valve is a break-away valve, a flapper valve, twist or screw type valve, or ruptures by breaking all ampoule.

20. The device of claim 1, wherein said mixing area comprises a reagent.

21. The device of claim 1, wherein said reagents are contained in a mixing area.

22. The device of claim 1, wherein said reagents are comprised in a solid substrate.

43. A kit comprising a sample collection device of claim 1 and a test device of claim 36, wherein each capture partner captures a different capture moiety.

23. A device comprising;

(a) a sample reservoir upstream of a plunger implement;

(b) a plurality of sealable apertures for delivery of one or more solutions into said device;

(c) a substrate for filtering one or more compounds, wherein said substrate or said device comprises reagents specific for at least one analyte.

24. A sample collection device comprising a body comprising:

(a) an upper sealed chamber containing buffer, and a valve;

(b) a lower chamber in fluid communication with the upper chamber containing

(c) a swab contained within said body;

(d) a mixing area downstream of said swab comprising an adsorbent or absorbent substrate, and at least one pair of reagents comprising a label and binding moiety.

25. A test device comprising a body comprising:

(a) a lateral flow membrane in the body; and

(b) upstream of the direction of lateral flow, a chamber comprising fluid, wherein the chamber is capable of controllably releasing the fluid into the lateral flow material, wherein there is a gap between said chamber and said lateral flow membrane.

26. The device of claim 25, wherein said gap is greater than zero and less than or equal to 3.0 mm.

27. The device of claim 25, wherein said gap is capable of closing if pressure is exerted onto said chamber.

28. The device of claim 25, wherein said chamber Contains a buffer solution with or without a detectable reagent.

29. The device of claim 28, wherein said detectable reagent is detectable by an instrument.

30. The device of claim 28 wherein said detectable reagent is detectable by an un-aided eye.

31. The device of claim 25, wherein said chamber is comprised of an ampoule.

32. The device of claim 25, wherein said controllably releasing comprises mechanical manipulation of said chamber.

33. The device of claim 25, wherein a cover or a flange provides a safeguard for said controllably releasing.

34. The device of claim 33, wherein said safeguard is broken or removed in order to gain access to said chamber.

35. The device of claim 25, wherein said membrane comprises one or more designate test and control regions.

50. A system comprising

(a) a test device of claim 25 or 36;

(b) a reader that reads a signal generated at any addressable line of the test device; and

(c) machine readable encoded instructions capable of directing said reader to detect the signal or signals.

51. The system of claim 50, wherein said device is adapted to be inserted into a reader.

52. The system of claim 51, wherein reader is adapted to allow said insertion only on condition that said chamber has controllably released said fluid.

61. The system of claim 50, wherein said label is europium.

36. A test device comprising a body comprising:

(a) a lateral flow membrane in the body and exposed through a single or a plurality of windows in the body wherein the membrane comprises;

(i) a wicking material upstream of said plurality of windows;

(ii) an absorbent material downstream of said plurality of windows;

(iii) a plurality of addressable lines, each line having immobilized thereto a capture partner, each capture partner capable of specifically binding to a different capture moiety; and

(iv) excluding any reagent specific to a target analyte.

37. The test device of claim 36, wherein the capture partners comprise oligonucleotide, avidin, streptavidin, pRNA, aptamer or a combination thereof.

38. The test device of claim 36, wherein said target analyte is an infectious agent.

39. The test device of claim 36, further comprising, upstream of the direction of lateral flow, a chamber comprising fluid that controllably releases the fluid into the wicking material.

40. The test device of claim 36, wherein the chamber is breakable or can be punctured.

41. The test device of claim 36, further comprising machine readable encoding instructions—

capable of initiating a protocol to detect—

signal from one or more addressable lines.

42. The device of claim 36, further comprising a chamber upstream of said membrane, wherein said chamber comprises a fluid solution.

44. A method comprising:

(a) forming a mixture by mixing a sample being tested for the presence of at least one target antigen with a solution comprising a plurality of reagents comprising;

(i) an antibody-capture moiety conjugate comprising an antibody which specifically binds a target antigen and a capture moiety, and

(ii) a labeled antibody, wherein the antibody specifically binds the same target antigen, wherein said reagents are directed against a plurality of different target antigens and form a complex;

(b) applying the mixture to a lateral flow membrane comprising a plurality of detection regions, each of said detection region having immobilized thereto a capture partner, each capture partner capable of specifically binding a different capture moiety;

(c) flowing the mixture across the membrane, whereby the capture moiety captures a complex having a capture moiety to which it is directed; and

(d) detecting label in at least one of said detection region, whereby detection of label indicates the presence of a target antigen in said sample.

45. The method of claim 44, wherein the capture partner is an oligonucleotide, avidin, streptavidin, pRNA, aptamer or a combination thereof.

46. The method of claim 44 wherein the label is a fluorophore, chromophore, metal, magnetic bead, raman particle or a combination thereof.

47. The method of claim 44, wherein said label is europium.

48. The method of claim 44, wherein said target antigens—

influenza viruses.

49. The method of claim 48, wherein said influenza viruses comprise—

influenza A and influenza B virus.

53. A system for detection of one or more analytes in a fluid sample, comprising:

(a) a sample collection device comprising;

(i) a swab sample collection implement,

(ii) a chamber containing fluid for extracting said swab,

(iii) and a plurality of pairs of first and second detection reagents, each pair capable of specifically binding a different target analyte in said fluid sample;

—

wherein either said first or second detection reagents further comprises a capture moiety—

; and

;

(b) a test device comprising a substrate defining a lateral flow path comprising a plurality of test regions positioned within said flow path, said test regions each containing an immobilized reagent capable of specifically binding to said capture moiety, wherein each of the test regions comprises a different immobilized reagent capable of binding a different capture moiety; and

at least one control region disposed in said lateral flow path, wherein said control region permits a determination of proper performance of the test device.

54. The system of claim 53, wherein said sample collection device further comprises a reservoir for collecting a portion of said fluid sample.

55. The system of claim 53, wherein said immobilized reagent is selected from the group consisting of an oligonucleotides, avidin, streptavidin, pyranosyl RNA (pRNA), aptamer, or a combination thereof.

56. The system of claim 53, wherein said capture moiety is selected from the group consisting of an oligonucleotides, avidin, streptavidin, pyranosyl RNA (pRNA), aptamer, or a combination thereof.

57. The system of claim 53, further comprising a reader into which said test device can be inserted, whereby said reader detects said one or more analytes.

58. The system of claim 53, wherein said plurality of first and second detection reagents are selected from the group consisting of an antibody, a nucleic acid, a receptor, a ligand and an antigen.

59. The system of claim 53, wherein said sample collection device is adapted to enable positive attachment to the test device and permit complete transfer of said specimen without spillage or contact with open air.

60. The system of claim 53, wherein said plurality of first detection reagents are labeled with a fluorophore, chromophore, metal or a combination thereof.

62. The system of claims 53, wherein said target analytes in said fluid sample comprise virus proteins and/or virus nucleic acids.

63. The system of claim 62, wherein said virus is influenza A and/or influenza B.

64. The system of claim 62, wherein said plurality of first and plurality of second reagents are antibodies.

65. The system of claim 63, wherein said influenza A includes subtypes of a formula HxNy, where x is, through 16, and y is 1 through 9, or any combination of xy thereof.

66. The system of claim 65, wherein said influenza A is H5N1.

67. The system of claim 53, wherein said test device further comprises a compartment upstream of said substrate.

68. The system of claim 67, wherein said compartment comprises a solution that is controllably released so as to come into contact with said substrate.

69. The system of claim 67, wherein said compartment and said substrate are separated by a gap.

70. The system of claim 68, wherein said solution is sealed in said compartment by a breakable seal.

71. The system of claim 70, wherein said seal is breakable by exerting pressure on said compartment.

72. A method for detection of one or more analytes in a fluid sample comprising:

(a) contacting a fluid sample with a system comprising;

(i) a sample collection implement comprising

(1) a swab,

(2) a chamber containing fluid for extracting said swab,

(3) a plurality of pairs of first and second detection reagents, each pair capable of specifically binding a different target analyte in said fluid sample;

wherein either said first or second detection reagents further comprises a capture moiety;

(ii) a test device comprising a substrate defining a lateral flow path comprising a plurality of test regions positioned within said flow path, said test regions each containing an immobilized reagent capable of specifically binding to said capture moiety, wherein each of the test regions comprises a different immobilized reagent capable of binding a different capture moiety; and

at least one control region disposed in said lateral flow path, wherein said control region permits a determination of proper performance of the sensor;

(b) detecting whether one or more analytes bind to said immobilized capturing reagent, disposed in said test device; and

(c) determining whether one or more analytes are present in said fluid sample, if a signal is detected in at least one of said plurality of test regions.

73. The method of claim 72, wherein said sample collection implement further comprises a reservoir for preserving a portion of said sample.

74. The method of claim 72, where said result remains stable for visualization for a time period greater than 30 minutes.

75. The method of claim 72, wherein said system'"'"'s sensitivity, based on dilution, is equivalent to 2500 copies of Influenza nucleic acid and specificity is 98% or greater.

76. The method of claim 72, wherein said immobilized reagent is selected from the group consisting of an oligonucleotide, avidin, streptavidin, pRNA, aptamer or a combination thereof.

77. The method of claim 72, wherein said immobilized capturing agent is pRNA.

78. The method of claim 72, wherein each reagent in said plurality of first detection reagents comprises a label.

79. The method of claim 78, wherein said label is selected from a group consisting of a fluorophore, a chromophore, a microparticle, a metal or a combination thereof.

80. The method of claim 78, wherein said label is europium.

81. The method of claim 72, wherein said multiple one or more analytes are one or more viruses.

82. The method of claim 81, wherein said one or more viruses include influenza A and/or influenza B strain.

83. The method of claim 81, wherein said one or more viruses include different virus strains and different virus subtypes.

84. The method of claim 83, wherein said different strains or said different subtypes include, pandemic virus, a non-pandemic virus, vaccine-available virus, vaccine-unavailable virus, or a combination thereof.

85. The method of 72, wherein each of said plurality of test regions comprises said immobilized reagent specific for said capture.

86. A method for determining whether a subject is infected with one or more types of influenza virus and/or subtypes and/or strains, comprising

(a) contacting a sample from said subject with a system capable of detecting multiple analytes, wherein said system comprises components that are capable of specifically binding a component of one or more of said influenza virus strains and/or subtypes thus forming one or more complexes;

wherein complexes formed from said—

sample travel through said system via a lateral flow path, wherein said one or more complexes are capable of specifically binding an immobilized capture reagent present in two or more distinct test regions along said lateral flow path, and wherein each of said two or more test regions comprises a different said immobilized capture reagent capable of capturing a different analyte complex;

(b) determining whether said one or more influenza virus strains and subtypes are present in said fluid sample by observing said two or more test regions, wherein a positive result indicates presence of a specific Influenza virus type and/or subtype and said positive result is stable for a period greater than 30 minutes without change.

87. A method for determining if a patient is infected with a specific Type of Influenza virus and/or subtype comprising:

(a) collecting a sample from said patient utilizing a sample collection device comprising;

(i) a plurality of immunoreagents that are capable of detecting different virus Types and subtypes, wherein for any given said virus Type or subtype said sample collection device contains a pair of said immunoreagents specific for a particular said virus Type or subtype, where members of the pair comprise a capture moiety and label respectively;

(b) inserting said sample collection device into a lateral flow test device, where said insertion provides a closed fluid communication between said sample collection device and said lateral flow test device, wherein said test device comprises one more test regions, wherein each of which said test regions comprises a particular immobilized capture reagent that is specific to one of said capture moiety in (a);

(c) detecting said label at one or more said test regions to identify which of said virus Types or subtypes are present in said sample; and

thereby determining whether a patient is infected with one or more influenza virus Types and/or subtypes.

89. The method of claim 87, wherein said label for any given said pair is selected from the group consisting of a fluorophore, chromophore, metal or combination thereof.

90. The method of claim 87, wherein said label is europium.

91. The method of claim 87, wherein said capture moiety is selected from the group consisting of an oligonucleotide, aptamer, pRNA, avidin, streptavidin, or a combination thereof.

92. The method of claim 87, wherein said capture moiety is pRNA.

88. The method of claim 88, further comprising determining the presence of influenza Type A virus of one or more subtypes selected from a pandemic, a non-pandemic, a subtype for which there are vaccines available and a subtype for which there are no vaccines available.

93. A method for processing a sample from a subject comprising;

(a) collecting a sample from said subject with a sample collection device comprising one or more identical identity tags, wherein said sample collection device further comprises an archival reservoir for preserving at least a portion of said sample;

(b) assaying in a test device said sample to detect one or more analytes by providing a detectable result on said test device;

(c) attaching one of said identity tags to each of said archival reservoir and said test device, at any time before or after steps a or b; and

thereby processing said sample from said subject.

94. The method of claim 93, wherein said identity tag is selected from a barcode, radio frequency emitter, light energy emitter, electromagnetic wave emitter, or a combination thereof.

95. The method of claim 93, wherein said sample collection device further comprises a sample collection implement and said method further comprises attaching an identical identity tag to said implement.

96. A lateral flow test device for the determination of one or more analytes comprising a test strip having one or more discrete bands of pRNA non-diffusively bound for the indirect specific capture of the one or more analytes.

97. The device of claim 96 farther comprising a control band non-diffusively bound to said strip.

98. The device of claim 96 wherein said pRNA is bound to said strip through a hydrophilic protein.

99. The device of claim 96 wherein said strip includes a detection zone having multiple discrete bands of pRNA each capable of specifically capturing and identifying a different analyte and a control zone downstream of said detection zone having a band adapted to demonstrate that a liquid sample has migrated through said detection zone.

100. The device of claim 99 further comprising a sample application zone upstream of said detection zone.

101. A method of detecting one or more analytes in a fluid sample comprising applying said sample to a lateral flow device which comprises one or more discrete bands of pRNA non-diffusively bound to said device, which pRNAs are capable of indirectly specifically binding to said one or more analytes.

102. The method of claim 101 further comprising prior to application to said device reacting said sample with one first conjugate for each of said analytes, each said conjugate comprising a specific binding partner capable of specifically binding to an analyte and a pRNA adapted to recognize one of said discrete bands of pRNA on said device.

103. The method of claim 102 further comprising adding a second conjugate for each said analyte comprising a specific binding partner capable of binding to said analyte and a label.

104. The method of claim 103 wherein said second conjugate is added to said sample together with the first conjugate before application of the mixture to said device or applying said second conjugate to said device after the application of said first conjugate.

105. The method of claim 101 wherein said device comprises a bibulous strip having a sample application zone, upstream of a detection zone containing said bands of pRNA and a control zone downstream of said detection zone.

107. The pRNA of claims 96, 101 or 106, having any of one or more sequences provided in Table 5.

106. A kit comprising:

(a) a lateral flow device comprising a bibulous strip containing (i) a sample application zone, (ii) a detection zone comprising one or more discrete bands of pRNA non-diffusively bound to said strip, each band capable of indirectly specifically capturing a different analyte and (iii) a control zone;

(b) a first conjugate for each of said analytes, each said conjugate comprising a specific binding partner capable of specifically binding to an analyte and a pRNA adapted to recognize one of said discrete bands of pRNA on said strip; and

(c) a second conjugate for each of said analytes comprising a specific binding partner capable of specifically binding to said analyte and a label.

108. A method of detecting one or more analytes in a sample obtained from a subject comprising applying said sample to an assay system, wherein said system comprises at least one pair of complementary pRNA sequences, wherein one member of said at least one pair is conjugated to an antibody that is specific for said one or more analytes.

109. The method of claim 108, wherein said one or more analytes are proteins from an infectious agent.

110. The method of claim 109, wherein said infectious agent is influenza virus, HIV, hepatitis virus, adenovirus, enterovirus or parainfluenza virus.

111. The method of claim 108, wherein said one or more analytes comprise at least two different types of an infectious agent.

112. The method of claim 108, wherein said one or more analytes comprise at least three different types, subtypes or a combination of types and subtypes of an infectious agent.

113. The method of claim 111 or 112, wherein said infectious agent is influenza virus.

114. The method of claim 113, wherein said influenza virus is selected from influenza A, B, C and a combination thereof.

115. The method of claim 113, wherein said influenza virus is selected from the group consisting of influenza A, influenza B, a subtype of influenza A that is pandemic, and a subtype of influenza A that is non-pandemic.

116. The method of claim 113, wherein said influenza virus is H5N1 subtype.

117. A device configured to detect a test signal from a test sample, wherein said device is also configured to detect a reference signal, wherein said reference signal is permanent and integrated into said device.

118. The device of claim 117, wherein said test and reference signals is emitted by a fluorescence or phosphorescent label.

119. The device of claim 118, wherein said label comprises a lanthanide metal.

120. The device of claim 118, wherein said label comprises europium, samarium, terbium or dysprosium.

121. The device of claim 117, wherein said label is Europium III.

122. The device of claim 117 or 122, wherein said label is embedded or cast in a polymer material.

123. The device of claim 123, wherein said polymer comprises an acrylic, glass, plastic, rubber or vinyl material.

124. The device of claim 124, wherein said polymer comprises acrylic.

125. The fluorescence reader comprising a hard standard, wherein said reader is configured to read a test sample containing an analyte labeled with a molecule, wherein said hard standard and said label have large Stoke'"'"'s shifts.

126. The reader of claim 126, wherein said hard standard and said label comprise a lanthanide.

127. The reader of claim 126, wherein said hard standard and said label are selected from europium, samarium terbium and dysprosium.

128. The reader of claim 126, wherein said reader comprise at least two hard standards.

129. A UV reader comprising at least one permanent reference standard signal.

130. The reader of claim 130, wherein said reader comprises a laser or photodiode.

131. The reader of claim 130, wherein said reader said reference signal is a fluorescence signal.

133. The reader of claims 130, wherein said signal is by Europium.

134. The reader of claim 133, wherein said reader is configured with a light emitting diode that emits in the UV spectrum.

137. The reader of claim 133, wherein said reader is configured with a light emitting diode that emits light primarily in a range from between about 390 nm to 425 nm.

138. The reader of claim 133, wherein said reader is an LED reader.

132. The reader of claim 132, wherein said fluorescence is by a lanthanide.

135. The reader of claim 135, wherein said reader further comprises one or more filter(s) that limits excitation light.

136. The reader of claim 136, wherein said one or more filter(s) is selected from a short pass, band pass, long pass, and a combination thereof.

139. A UV LED reader configured to detect a fluorescence signal from a test strip.

140. The reader of claim 140, wherein said fluorescence signal is emitted from a lanthanide.

141. The reader of claim 140, wherein said fluorescence signal is emitted from Europium.

142. The reader of claim 140, wherein said reader is further adapted to comprise at least one hard reference standard.

143. The reader of claim 140, wherein said reader comprise a first and second hard reference standard.

144. The reader of claim 140, wherein said reader comprises a photodiode that emits in a visible portion of a spectrum.

145. The reader of claim 140, wherein said reader further comprises one or more filter(s) that limit excitation light.

146. The reader of claim 146, wherein said one or more filter(s) is selected from a short pass, band pass, long pass and a combination thereof.

147. The reader of claim 146, wherein said reader is configured to comprise an axis of excitation light that is at least at a 90 degree angle relative to said test strip.

148. The reader of claim 146, wherein said reader is configured to comprise an axis of emitted light that is at an angle other than 90 degrees relative to said test strip.

149. A fluorescence reader adapted for specifically and sensitively measuring the emissions of a fluorescent material comprising:

(a) a light emitting diode which emits in the UV region of the spectrum

(b) a photodiode capable of detecting the emitted fluorescent signal.

150. The reader of claim 150 adapted to measure the fluorescent material on a lateral flow test strip.

151. The reader of claim 150, wherein said light emitting diode and photodiode are positioned so the axis of the excitation light from said light emitting diode is at a 90 degree angle to said test strip and said the axis at which said photodiode receives the emitted light is at an angle other than 90 degrees to said test strip.

152. The reader of claim 150, wherein the wavelengths of said light emitting diode and photodiode are optimized for detecting Europium.

153. A system for the detection of one or more analytes comprising:

(a) a lateral flow test strip comprising one or more discrete bands of a specific binding partner capable of directly or indirectly binding said one or more analytes; and

(b) a fluorescence reader capable of distinguishing a fluorescent signal from each of the one or more bands on said test strip, said reader comprising a light emitting diode which emits in the UV region of the spectrum and a photodiode capable of detecting the emitted fluorescent signal.

154. The system of claim 154 wherein said fluorescent signal is generated by Europium.