CA2578313A1 - Highly accurate rapid parallel immunoassay device - Google Patents
Highly accurate rapid parallel immunoassay device Download PDFInfo
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- CA2578313A1 CA2578313A1 CA 2578313 CA2578313A CA2578313A1 CA 2578313 A1 CA2578313 A1 CA 2578313A1 CA 2578313 CA2578313 CA 2578313 CA 2578313 A CA2578313 A CA 2578313A CA 2578313 A1 CA2578313 A1 CA 2578313A1
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- 238000003018 immunoassay Methods 0.000 title 1
- 239000000427 antigen Substances 0.000 claims abstract description 54
- 102000036639 antigens Human genes 0.000 claims abstract description 54
- 108091007433 antigens Proteins 0.000 claims abstract description 54
- 201000010099 disease Diseases 0.000 claims abstract description 36
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 36
- 239000003550 marker Substances 0.000 claims abstract description 21
- 238000012360 testing method Methods 0.000 claims description 77
- 239000012528 membrane Substances 0.000 claims description 35
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 17
- 210000004369 blood Anatomy 0.000 claims description 9
- 239000008280 blood Substances 0.000 claims description 9
- 210000002966 serum Anatomy 0.000 claims description 8
- 210000001124 body fluid Anatomy 0.000 claims description 2
- 210000003296 saliva Anatomy 0.000 claims description 2
- 210000002700 urine Anatomy 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 claims 1
- 241000700605 Viruses Species 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 claims 1
- 210000004927 skin cell Anatomy 0.000 claims 1
- 238000002405 diagnostic procedure Methods 0.000 abstract description 5
- 238000001514 detection method Methods 0.000 abstract description 3
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 10
- 241000725303 Human immunodeficiency virus Species 0.000 description 8
- 239000002250 absorbent Substances 0.000 description 7
- 230000002745 absorbent Effects 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 239000000463 material Substances 0.000 description 6
- 230000035945 sensitivity Effects 0.000 description 6
- 239000000872 buffer Substances 0.000 description 5
- 230000003100 immobilizing effect Effects 0.000 description 5
- 241000713340 Human immunodeficiency virus 2 Species 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 208000031886 HIV Infections Diseases 0.000 description 3
- 101800000385 Transmembrane protein Proteins 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 230000000007 visual effect Effects 0.000 description 3
- 208000035473 Communicable disease Diseases 0.000 description 2
- 208000037357 HIV infectious disease Diseases 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 description 2
- 238000012125 lateral flow test Methods 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000008447 perception Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
- G01N33/56988—HIV or HTLV
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/15—Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
- G01N2333/155—Lentiviridae, e.g. visna-maedi virus, equine infectious virus, FIV, SIV
- G01N2333/16—HIV-1, HIV-2
- G01N2333/162—HIV-1, HIV-2 env, e.g. gp160, gp110/120, gp41, V3, peptid T, DC4-Binding site
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- Tropical Medicine & Parasitology (AREA)
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Abstract
The present invention relates to diagnostic test kit designs comprising distinctly applied, multiple recombinant antigens, of different epitopes of the same disease marker, that are capable of accurate, parallel detection of the presence of disease specific antibodies in a given specimen.
Description
DESCRIPTION
1. Background of the Invention The rapid tests' ability to produce accurate and reliable results depends not only on the intrinsic quality of the tests themselves but also on the extrinsic factors such as the quality/type of biological specimens, the ability of the user to correctly perform the tests, and the prevalence rate of the target disease in a given population. The probability that a single rapid test will accurately determine the true infection status of a person varies with the prevalence of the infectious disease in that population.
Generally, when there's a higher prevalence of an infectious disease in a population, there's also a greater probability that a person from that population testing positive is truly infected (i.e. the greater the positive predictive value). This means that the proportion of biological specimens testing false-positive for a given disease decreases in high prevalence settings. Similarly, specimens showing negative test results in high prevalence settings may have a higher likelihood of being falsely negative (i.e. negative predictive value).
According to World Health Organization and UNAIDS' recommendations, published on 21 March 1997, in the context of rapid antibody testing for HIV diagnosis, while taking into consideration of different prevalence of infection settings, three disease testing strategies have been presented to statistically improve the accuracy of the test results while minimizing the cost. These strategies are again published by WHO/UNAIDS in 2001 in "Guidelines for Using HIV Testing Technologies in Surveillance". The strategies are described below.
Strategy I: Requires one test; for use in diagnostic testing in populations with an HIV prevalence >30% among persons with clinical signs or symptoms of HIV infection; for use in blood screening, for all prevalence rates; and for use in surveillance testing in populations with an HIV prevalence >10%.
Strategy II: Requires up to two tests; for use in diagnostic testing in populations with an HIV
prevalence 530% among persons with clinical signs or symptoms of HIV infection or >10% among asymptomatic persons; and for use in surveillance testing in populations with an HIV prevalence <_10%.
Strategy III: Requires up to three tests; and for use in diagnostic testing in populations with an HIV
prevalence 510% among asymptomatic persons.
Strategies 11 and 11I, see Fig. 9, can be performed in serial or in parallel.
WHO/UNAIDS adopts the serial testing algorithms with the concem that parallel testing algorithms require higher cost as two or more tests are always used simultaneously. Serial testing may be lengthy and may require more than one specimen collection. Advantages for performing parallel testing include:
Reduction in the risk of false negative results in high prevalence populations; reduction in the risk of false positive results in low prevalence populations; require only a single specimen, avoiding the collection of additional specimens;
favorable perception that two or more tests are better than one; and reduction in the stigma of the patient being called back for the second test. It is also noted in WHO/UNAIDS' recommendations that the tests used for the above testing algorithms should contain different antigens. Fig. 8 shows a parallel testing algorithm for HIV (source: Global AIDS Program, Centers for Disease Control and Prevention, USA).
1. Background of the Invention The rapid tests' ability to produce accurate and reliable results depends not only on the intrinsic quality of the tests themselves but also on the extrinsic factors such as the quality/type of biological specimens, the ability of the user to correctly perform the tests, and the prevalence rate of the target disease in a given population. The probability that a single rapid test will accurately determine the true infection status of a person varies with the prevalence of the infectious disease in that population.
Generally, when there's a higher prevalence of an infectious disease in a population, there's also a greater probability that a person from that population testing positive is truly infected (i.e. the greater the positive predictive value). This means that the proportion of biological specimens testing false-positive for a given disease decreases in high prevalence settings. Similarly, specimens showing negative test results in high prevalence settings may have a higher likelihood of being falsely negative (i.e. negative predictive value).
According to World Health Organization and UNAIDS' recommendations, published on 21 March 1997, in the context of rapid antibody testing for HIV diagnosis, while taking into consideration of different prevalence of infection settings, three disease testing strategies have been presented to statistically improve the accuracy of the test results while minimizing the cost. These strategies are again published by WHO/UNAIDS in 2001 in "Guidelines for Using HIV Testing Technologies in Surveillance". The strategies are described below.
Strategy I: Requires one test; for use in diagnostic testing in populations with an HIV prevalence >30% among persons with clinical signs or symptoms of HIV infection; for use in blood screening, for all prevalence rates; and for use in surveillance testing in populations with an HIV prevalence >10%.
Strategy II: Requires up to two tests; for use in diagnostic testing in populations with an HIV
prevalence 530% among persons with clinical signs or symptoms of HIV infection or >10% among asymptomatic persons; and for use in surveillance testing in populations with an HIV prevalence <_10%.
Strategy III: Requires up to three tests; and for use in diagnostic testing in populations with an HIV
prevalence 510% among asymptomatic persons.
Strategies 11 and 11I, see Fig. 9, can be performed in serial or in parallel.
WHO/UNAIDS adopts the serial testing algorithms with the concem that parallel testing algorithms require higher cost as two or more tests are always used simultaneously. Serial testing may be lengthy and may require more than one specimen collection. Advantages for performing parallel testing include:
Reduction in the risk of false negative results in high prevalence populations; reduction in the risk of false positive results in low prevalence populations; require only a single specimen, avoiding the collection of additional specimens;
favorable perception that two or more tests are better than one; and reduction in the stigma of the patient being called back for the second test. It is also noted in WHO/UNAIDS' recommendations that the tests used for the above testing algorithms should contain different antigens. Fig. 8 shows a parallel testing algorithm for HIV (source: Global AIDS Program, Centers for Disease Control and Prevention, USA).
2. Summary of the Invention In light of the above strategies published by WHO/UNAIDS and CDC's parallel testing algorithm, the present invention relates to diagnostic test kit designs comprising multiple recombinant antigens, of different epitopes of the same disease marker, that are capable of accurate, simultaneous, parallel detection of the presence of disease specific antibodies in a single collection of specimen applied to a single test device, thus avoiding the need to perform multiple rapid tests or to repeatedly collect specimens. The present invention therefore greatly improves the accuracy of current immunoconcentration (flow-through) and immunochromatographic (lateral-flow) rapid tests.
Flow-through tests employ solid-phase capture technology, which involves the immobilization of antigens on a porous membrane. The specimen flows through the membrane and is absorbed into an absorbent material on the opposite side. A dot or a line visibly forms on the membrane when developed with a signal reagent (usually a colloidal gold or selenium conjugate). Some tests allow the detection of multiple diseases or disease subtypes by immobilizing antigens from disease specific markers to different locations on the membrane. One such example is Blo-Rad Laboratories' MultispotTM' HIV-1/HIV-2 Rapid Test. The flow-through tests usually require a few steps for the addition of specimen, wash buffer, and signal reagent. Several flow-through devices include a procedural control on the membrane; the appearance of a colored dot or line at this location confirms the test has been performed correctly. US 20030165970 and WO 03098215 describe examples of flow-through devices.
Lateral-flow strips incorporate both antigen and signal reagent into a strip of porous membrane and absorbent materials. The specimen, sometimes followed by a buffer, is applied to the absorbent/sampling area of the device. Alternatively, the specimen is diluted in a vial of buffer, into which the test device is inserted or from which a quantity is drawn out and applied to the test device.
The specimen combines with the signal reagent and laterally migrates through the porous membrane. A
positive reaction results in a visual line on the membrane where antigens have been applied. A
procedural control line is usually applied to the strip at a location beyond the antigen line. A visual line at both the test and control locations indicates a positive test result, a line only at the control location indicates a negative test result, and the absence of a line at the control location means the test is invalid.
Some tests apply different antigens in different locations and allow differentiation of antibodies to two or more disease specific antibodies. In most lateral-flow devices, the test strip is encased in a plastic cartridge. EP 0306772, GB 2204398, EP 38619, EP 0225054, EP 0183442, and EP
0186799 describe examples of lateral-flow devices.
Flow-through tests employ solid-phase capture technology, which involves the immobilization of antigens on a porous membrane. The specimen flows through the membrane and is absorbed into an absorbent material on the opposite side. A dot or a line visibly forms on the membrane when developed with a signal reagent (usually a colloidal gold or selenium conjugate). Some tests allow the detection of multiple diseases or disease subtypes by immobilizing antigens from disease specific markers to different locations on the membrane. One such example is Blo-Rad Laboratories' MultispotTM' HIV-1/HIV-2 Rapid Test. The flow-through tests usually require a few steps for the addition of specimen, wash buffer, and signal reagent. Several flow-through devices include a procedural control on the membrane; the appearance of a colored dot or line at this location confirms the test has been performed correctly. US 20030165970 and WO 03098215 describe examples of flow-through devices.
Lateral-flow strips incorporate both antigen and signal reagent into a strip of porous membrane and absorbent materials. The specimen, sometimes followed by a buffer, is applied to the absorbent/sampling area of the device. Alternatively, the specimen is diluted in a vial of buffer, into which the test device is inserted or from which a quantity is drawn out and applied to the test device.
The specimen combines with the signal reagent and laterally migrates through the porous membrane. A
positive reaction results in a visual line on the membrane where antigens have been applied. A
procedural control line is usually applied to the strip at a location beyond the antigen line. A visual line at both the test and control locations indicates a positive test result, a line only at the control location indicates a negative test result, and the absence of a line at the control location means the test is invalid.
Some tests apply different antigens in different locations and allow differentiation of antibodies to two or more disease specific antibodies. In most lateral-flow devices, the test strip is encased in a plastic cartridge. EP 0306772, GB 2204398, EP 38619, EP 0225054, EP 0183442, and EP
0186799 describe examples of lateral-flow devices.
3. Brief Description of the Drawings Fig. 1 illustrates an example of a flow-through rapid test device.
Fig. 2A & Fig. 2B illustrate examples of immobilizing a procedural control and three antigens of different epitopes of the same disease marker on four different locations on the same membrane in a single flow-through rapid test device.
Fig. 3A & Fig. 3B illustrate examples of immobilizing a procedural control and two antigens of different epitopes of the same disease marker on three different locations on the same membrane in a single flow-through rapid test device.
Fig. 4 illustrates an example of a lateral-flow rapid test device.
Fig. 5A & Fig. 5B illustrate examples of immobilizing two antigens of different epitopes of the same disease marker on two different locations on the same membrane or two separate membranes in a single lateral-flow rapid test device.
Fig. 6 illustrates an example of immobilizing antigens of different epitopes of the same disease marker on 2 separate membranes in a single lateral-flow rapid test device.
Fig. 7A & Fig. 7B illustrate examples of test strips that are housed within the lateral-flow rapid test device in Fig. 6.
Fig. 8 is a diagram of a parallel testing algorithm sourced from Global AIDS
Program, Centers for Disease Control and Prevention, USA.
Fig. 9 is a diagram of testing strategies adopted by of WHO/UNAIDS.
Fig. 2A & Fig. 2B illustrate examples of immobilizing a procedural control and three antigens of different epitopes of the same disease marker on four different locations on the same membrane in a single flow-through rapid test device.
Fig. 3A & Fig. 3B illustrate examples of immobilizing a procedural control and two antigens of different epitopes of the same disease marker on three different locations on the same membrane in a single flow-through rapid test device.
Fig. 4 illustrates an example of a lateral-flow rapid test device.
Fig. 5A & Fig. 5B illustrate examples of immobilizing two antigens of different epitopes of the same disease marker on two different locations on the same membrane or two separate membranes in a single lateral-flow rapid test device.
Fig. 6 illustrates an example of immobilizing antigens of different epitopes of the same disease marker on 2 separate membranes in a single lateral-flow rapid test device.
Fig. 7A & Fig. 7B illustrate examples of test strips that are housed within the lateral-flow rapid test device in Fig. 6.
Fig. 8 is a diagram of a parallel testing algorithm sourced from Global AIDS
Program, Centers for Disease Control and Prevention, USA.
Fig. 9 is a diagram of testing strategies adopted by of WHO/UNAIDS.
4. Detailed Description of the Invention Referring to Fig. 1, an example of a flow-through rapid test device with a solid casing 4 composed of a well-shaped receptacle 2 into which specimen fluid and complementary buffers and reagents can be poured, a porous membrane 1 immediately at the bottom of the receptacle that contains an immobilized procedural control and antigens of different epitopes of the same disease marker, and a reservoir 3 within which holds an absorbent material meant to collect and retain fluids that are flown through the membrane 1 and to support the membrane.
According to one embodiment of the present invention, the flow-through rapid test device's porous membrane 1 contains three immobilized antigens and one procedural control.
Fig. 2A and Fig. 2B show two examples of possible arrangements of the immobilized antigens and procedural control at four locations on the porous membrane 1. One of the four locations at 6, 7, 8, or 9 shown in Fig. 2A contains a quantity of immobilized procedural control. Each of the three remaining locations, other than the location containing the immobilized procedural control, contains a quantity of immobilized antigens.
Each of the three immobilized antigens locations contains different recombinant antigens of a different epitope of the same disease marker. The procedural control and/or each of the three different antigens can be applied at one of the four locations as a dot, line, or any visually distinguishable shapes.
According to another embodiment of the present invention, the flow-through rapid test device's porous membrane 1 contains two immobilized antigens and one procedural control. Fig.
3A and Fig. 3B show two examples of possible arrangement of the immobilized antigens and procedural control at three locations on the porous membrane 1. One of the three locations at 10, 11, or 12 shown in Fig. 3A
contains a quantity of immobilized procedural control. Each of the two remaining locations, other than the location containing the immobilized procedural control, contains a quantity of immobilized antigens.
Each of the two immobilized antigen locations contains different recombinant antigens of a different epitope of the same disease marker. The procedural control and/or each of the two different antigens can be applied at one of the three locations as a dot, line, or any visually distinguishable shapes.
The following examples are intended to illustrate the benefits of the present invention, but do not exemplify the full scope of the invention. For example, although gp4l from Human Immunodeficiency Virus Type 1 and gp36 from Human Immunodeficiency Virus Type 2 are exclusively exemplified as disease markers, a variety of other disease makers are also applicable.
Similarly, EPITOPE1 antigen, with an amino acid sequence SEQID1, is exclusively exemplified as one epitope capable of capturing/binding with antibodies that are specific to gp4l, but a variety of other recombinant antigens with different amino acid sequences are also applicable, so long as they are capable of capturing/binding with antibodies that are specific to gp4l. Furthermore, serum, plasma, and whole blood are exclusively exemplified as specimen types, but a variety of other bodily fluids and tissues, not limited to urine and saliva, are also applicable.
Example 1 According to this example, the flow-through test device contains two different recombinant antigens;
each of the two different antigens has a different epitope of the disease marker gp4l from Human Immunodeficiency Virus Type 1. Both epitopes are capable of capturing antibodies specific to gp4l in an HIV-1 infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. One of the two antigens has an epitope (EPITOPE1) with an amino acid sequence SEQID1.
The other has an epitope (EPITOPE2) with an amino acid sequence SEQID2. The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 3A, where the procedural control is immobilized at 10, EPITOPEI is immobilized at 11, and EPITOPE2 is immobilized at 12. Fig. 3B shows another possible arrangement of the three locations.
Example 2 According to this example, the same flow-through test device in Example 1 simultaneously contains immobilized EPITOPE4 and EPITOPE5, at the same locations as EPITOPE1 and respectively. EPITOPE4 and EPITOPE5 are capable of capturing antibodies, which can bind to the disease marker gp36 from Human Immunodeficiency Virus Type 2, in an infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. EPITOPE4 has an amino acid sequence SEQID4. EPITOPE5 has an amino acid sequence SEQID5.
Example 3 According to this example, the flow-through test device contains three different recombinant antigens;
each of the three different antigens has a different epitope of the disease marker gp4l from Human Immunodeficiency Virus Type 1. All three epitopes are capable of capturing antibodies specific to gp4l in an HIV-1 infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. One of the three antigens has an epitope (EPITOPE1) with an amino acid sequence SEQID1. The other has an epitope (EPITOPE2) with an amino acid sequence SEQID2. The last has an epitope (EPITOPE3) with an amino acid sequence SEQID3. The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 2A, where the procedural control is immobilized at 6, EPITOPE1 is immobilized at 7, EPITOPE2 is immobilized at 8, and EPITOPE3 is immobilized at 9. Fig. 2B shows another possible arrangement of the four locations.
Example 4 According to this example, the same flow-through test device in Example 3 simultaneously contains immobilized EPITOPE4, EPITOPE5, and EPITOPE6, at the same location as EPITOPEI, EPITOPE2, and EPITOPE3 respectively on the porous membrane. EPITOPE4, EPITOPE5, and EPITOPE6 are capable of capturing antibodies, which can bind to the disease marker gp36 from Human Immunodeficiency Virus Type 2, in an infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. EPITOPE4 has an amino acid sequence SEQID4.
EPITOPE5 has an amino acid sequence SEQID5. EPTIOPE6 has an amino acid sequence SEQID6.
According to one embodiment of the present invention, the flow-through rapid test device's porous membrane 1 contains three immobilized antigens and one procedural control.
Fig. 2A and Fig. 2B show two examples of possible arrangements of the immobilized antigens and procedural control at four locations on the porous membrane 1. One of the four locations at 6, 7, 8, or 9 shown in Fig. 2A contains a quantity of immobilized procedural control. Each of the three remaining locations, other than the location containing the immobilized procedural control, contains a quantity of immobilized antigens.
Each of the three immobilized antigens locations contains different recombinant antigens of a different epitope of the same disease marker. The procedural control and/or each of the three different antigens can be applied at one of the four locations as a dot, line, or any visually distinguishable shapes.
According to another embodiment of the present invention, the flow-through rapid test device's porous membrane 1 contains two immobilized antigens and one procedural control. Fig.
3A and Fig. 3B show two examples of possible arrangement of the immobilized antigens and procedural control at three locations on the porous membrane 1. One of the three locations at 10, 11, or 12 shown in Fig. 3A
contains a quantity of immobilized procedural control. Each of the two remaining locations, other than the location containing the immobilized procedural control, contains a quantity of immobilized antigens.
Each of the two immobilized antigen locations contains different recombinant antigens of a different epitope of the same disease marker. The procedural control and/or each of the two different antigens can be applied at one of the three locations as a dot, line, or any visually distinguishable shapes.
The following examples are intended to illustrate the benefits of the present invention, but do not exemplify the full scope of the invention. For example, although gp4l from Human Immunodeficiency Virus Type 1 and gp36 from Human Immunodeficiency Virus Type 2 are exclusively exemplified as disease markers, a variety of other disease makers are also applicable.
Similarly, EPITOPE1 antigen, with an amino acid sequence SEQID1, is exclusively exemplified as one epitope capable of capturing/binding with antibodies that are specific to gp4l, but a variety of other recombinant antigens with different amino acid sequences are also applicable, so long as they are capable of capturing/binding with antibodies that are specific to gp4l. Furthermore, serum, plasma, and whole blood are exclusively exemplified as specimen types, but a variety of other bodily fluids and tissues, not limited to urine and saliva, are also applicable.
Example 1 According to this example, the flow-through test device contains two different recombinant antigens;
each of the two different antigens has a different epitope of the disease marker gp4l from Human Immunodeficiency Virus Type 1. Both epitopes are capable of capturing antibodies specific to gp4l in an HIV-1 infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. One of the two antigens has an epitope (EPITOPE1) with an amino acid sequence SEQID1.
The other has an epitope (EPITOPE2) with an amino acid sequence SEQID2. The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 3A, where the procedural control is immobilized at 10, EPITOPEI is immobilized at 11, and EPITOPE2 is immobilized at 12. Fig. 3B shows another possible arrangement of the three locations.
Example 2 According to this example, the same flow-through test device in Example 1 simultaneously contains immobilized EPITOPE4 and EPITOPE5, at the same locations as EPITOPE1 and respectively. EPITOPE4 and EPITOPE5 are capable of capturing antibodies, which can bind to the disease marker gp36 from Human Immunodeficiency Virus Type 2, in an infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. EPITOPE4 has an amino acid sequence SEQID4. EPITOPE5 has an amino acid sequence SEQID5.
Example 3 According to this example, the flow-through test device contains three different recombinant antigens;
each of the three different antigens has a different epitope of the disease marker gp4l from Human Immunodeficiency Virus Type 1. All three epitopes are capable of capturing antibodies specific to gp4l in an HIV-1 infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. One of the three antigens has an epitope (EPITOPE1) with an amino acid sequence SEQID1. The other has an epitope (EPITOPE2) with an amino acid sequence SEQID2. The last has an epitope (EPITOPE3) with an amino acid sequence SEQID3. The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 2A, where the procedural control is immobilized at 6, EPITOPE1 is immobilized at 7, EPITOPE2 is immobilized at 8, and EPITOPE3 is immobilized at 9. Fig. 2B shows another possible arrangement of the four locations.
Example 4 According to this example, the same flow-through test device in Example 3 simultaneously contains immobilized EPITOPE4, EPITOPE5, and EPITOPE6, at the same location as EPITOPEI, EPITOPE2, and EPITOPE3 respectively on the porous membrane. EPITOPE4, EPITOPE5, and EPITOPE6 are capable of capturing antibodies, which can bind to the disease marker gp36 from Human Immunodeficiency Virus Type 2, in an infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. EPITOPE4 has an amino acid sequence SEQID4.
EPITOPE5 has an amino acid sequence SEQID5. EPTIOPE6 has an amino acid sequence SEQID6.
Now referring to Fig. 4, an example of lateral-flow rapid test device with a solid casing 14 that houses a test strip composed of absorbent materials and a porous membrane 13 that contains an immobilized procedural control and antigens of different epitopes of the same disease marker. The casing also has a receptacle 16 immediately above the absorbent materials 15 of the test strip into which specimen fluid and complementary buffers and reagents can be added, and a viewing window 17 immediately above the porous membrane 13 to allow visual reading of the testing result.
According to one embodiment of the present invention, the lateral-flow rapid test device's porous membrane 13 contains two immobilized antigens and one procedural control. Fig.
5A shows an example of possible arrangement of the immobilized antigens and procedural control at three locations 18, 19, and 20 on the porous membrane. One of the three locations shown in Fig. 5A
contains a quantity of immobilized procedural control. Each of the two remaining locations, other than the location containing the immobilized procedural control, contains a quantity of immobilized antigens. Each of the two immobilized antigens locations contains different recombinant antigens of a different epitope of the same disease marker. The procedural control and/or each of the two different antigens can be applied at one of the three locations as a dot, line, or any visually distinguishable shapes.
According to another embodiment of the present invention, as illustrated in Fig. 6, the modified lateral-flow rapid test has an additional porous membrane 21 containing one or more immobilized antigens and one optional procedural control. The additional membrane 21 can be supported on the same test strip Fig. 7A or on a separate test strip Fig. 7B housed within a single casing 14.
The casing has a single receptacle 16 immediately above the absorbent materials 15 of the test strip(s) into which specimen fluid and complementary buffers and reagents can be added, two viewing windows 17 and 22 immediately above the porous membrane 13 and 21, which contain immobilized procedural control and antigens of different epitopes of the same disease marker.
Example 5 According to this example, the lateral-flow test device contains two different recombinant antigens on the same porous membrane 13; each of the two different antigens has a different epitope of the disease marker gp4l from Human Immunodeficiency Virus Type 1. Both epitopes are capable of capturing antibodies specific to gp4l in an HIV-1 infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. One of the two antigens has an epitope (EPITOPE1) with an amino acid sequence SEQID1. The other has an epitope (EPITOPE2) with an amino acid sequence SEQID2. The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 5A, where the procedural control is immobilized at 18, EPITOPE1 is immobilized at 19, and EPITOPE2 is immobilized at 20.
According to one embodiment of the present invention, the lateral-flow rapid test device's porous membrane 13 contains two immobilized antigens and one procedural control. Fig.
5A shows an example of possible arrangement of the immobilized antigens and procedural control at three locations 18, 19, and 20 on the porous membrane. One of the three locations shown in Fig. 5A
contains a quantity of immobilized procedural control. Each of the two remaining locations, other than the location containing the immobilized procedural control, contains a quantity of immobilized antigens. Each of the two immobilized antigens locations contains different recombinant antigens of a different epitope of the same disease marker. The procedural control and/or each of the two different antigens can be applied at one of the three locations as a dot, line, or any visually distinguishable shapes.
According to another embodiment of the present invention, as illustrated in Fig. 6, the modified lateral-flow rapid test has an additional porous membrane 21 containing one or more immobilized antigens and one optional procedural control. The additional membrane 21 can be supported on the same test strip Fig. 7A or on a separate test strip Fig. 7B housed within a single casing 14.
The casing has a single receptacle 16 immediately above the absorbent materials 15 of the test strip(s) into which specimen fluid and complementary buffers and reagents can be added, two viewing windows 17 and 22 immediately above the porous membrane 13 and 21, which contain immobilized procedural control and antigens of different epitopes of the same disease marker.
Example 5 According to this example, the lateral-flow test device contains two different recombinant antigens on the same porous membrane 13; each of the two different antigens has a different epitope of the disease marker gp4l from Human Immunodeficiency Virus Type 1. Both epitopes are capable of capturing antibodies specific to gp4l in an HIV-1 infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. One of the two antigens has an epitope (EPITOPE1) with an amino acid sequence SEQID1. The other has an epitope (EPITOPE2) with an amino acid sequence SEQID2. The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 5A, where the procedural control is immobilized at 18, EPITOPE1 is immobilized at 19, and EPITOPE2 is immobilized at 20.
Example 6 According to this example, the lateral-flow test device contains two different recombinant antigens on two separate porous membranes 13 and 21; each of the two different antigens has a different epitope of the disease marker gp4l from Human Immunodeficiency Virus Type 1. Both epitopes are capable of capturing antibodies specific to gp4l in an HIV-1 infected patient's serum, plasma, or whole blood, but differ in sensitivity and specificity. One of the two antigens has an epitope (EPITOPE1) with an amino acid sequence SEQID1. The other has an epitope (EPITOPE2) with an amino acid sequence SEQID2.
The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 5A and 5B, where the procedural control is immobilized at 18 and 23, EPITOPE1 is immobilized at 19, and EPITOPE2 is immobilized at 22.
Numerous modifications and variations in practice of the invention are expected to occur to those skilled in the art upon consideration of the foregoing descriptions of preferred embodiments thereof. It is well within the skill in the art to practice the present invention accordingly to a wide variety of methods and formats. Consequently, only such limitations should be placed on the invention as appear in the following claims.
The arrangement of the locations on the porous membrane of the two immobilized antigens and the procedural control is similar to Fig. 5A and 5B, where the procedural control is immobilized at 18 and 23, EPITOPE1 is immobilized at 19, and EPITOPE2 is immobilized at 22.
Numerous modifications and variations in practice of the invention are expected to occur to those skilled in the art upon consideration of the foregoing descriptions of preferred embodiments thereof. It is well within the skill in the art to practice the present invention accordingly to a wide variety of methods and formats. Consequently, only such limitations should be placed on the invention as appear in the following claims.
Claims (7)
1. A rapid test device comprising a procedural control and two or more recombinant antigens, that are different epitopes of the same disease marker, applied at visually distinct locations on the porous membrane, to simultaneously capture antibodies that are specific to the disease marker in a single collected specimen.
2. The test device according to claim 1 is a flow-through rapid test.
3. The test device according to claim 1 is a lateral-flow rapid test.
4. The test device according to claim 1 wherein said epitopes can be partially identical to one another in terms of their amino acid sequences and/or surface structures.
5. The test device according to claim 1 wherein said disease marker is a protein molecule, which is introduced and/or generated by a disease-causing vector in the body.
6. According to claim 5 wherein said disease-causing vector includes, but is not limited to, virus and bacteria.
7. The test device according to claim 1 wherein said specimen means bodily fluids and tissues of animal or human origin; these include, but are not limited to, serum, plasma, whole blood, saliva, mucous, skin cells, and urine.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2578313 CA2578313A1 (en) | 2007-03-01 | 2007-03-01 | Highly accurate rapid parallel immunoassay device |
| PCT/CA2008/000391 WO2008104081A1 (en) | 2007-03-01 | 2008-02-29 | Parallel immunoassay device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2578313 CA2578313A1 (en) | 2007-03-01 | 2007-03-01 | Highly accurate rapid parallel immunoassay device |
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| CA2578313A1 true CA2578313A1 (en) | 2007-05-14 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2578313 Abandoned CA2578313A1 (en) | 2007-03-01 | 2007-03-01 | Highly accurate rapid parallel immunoassay device |
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| CA (1) | CA2578313A1 (en) |
| WO (1) | WO2008104081A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016035099A1 (en) * | 2014-09-05 | 2016-03-10 | Meril Diagnostics Private Limited | A flow through device for detection of multiple bioanalytes and a process thereof |
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| CN111837038A (en) * | 2018-02-01 | 2020-10-27 | 思迪亚生物科学公司 | Rapid quantitative assay for assessing duration of infection |
| US11011278B1 (en) | 2020-09-21 | 2021-05-18 | Biolytical Laboratories Inc. | Methods and rapid test kits facilitating epidemiological surveillance |
| CN113311159A (en) * | 2021-04-15 | 2021-08-27 | 南方医科大学 | Test strip for rapidly distinguishing recent and long-term HIV infection states through serum HIV-1 antibody detection and preparation method thereof |
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| AU7446501A (en) * | 2000-06-14 | 2001-12-24 | J.Mitra And Co. Ltd. | Diagnostic kit for invitro detection of hepatitis c |
| DE10108680A1 (en) * | 2001-02-23 | 2002-09-26 | Biognostic Ag | Test strips for the assay of biological materials, for clinical diagnosis etc. carry affinity molecules and can take or support a flow to give time-related reactions |
| CN1164949C (en) * | 2001-08-17 | 2004-09-01 | 上海数康生物科技有限公司 | Reagent case for synchronous detecting multiple kinds of infectious disease and its preparing method |
| CA2575852C (en) * | 2004-07-29 | 2013-03-26 | Siliang Zhou | Lateral flow system and assay |
| WO2006062800A1 (en) * | 2004-12-04 | 2006-06-15 | Freedom Health, Llc | Monoclonal and polyclonal antibodies to equine hemoglobin and apparatus and methods using the antibodies and/or peroxidase reactions in the identification and localization of ulcers in equines |
| CN1773285A (en) * | 2005-11-02 | 2006-05-17 | 中国农业科学院生物技术研究所 | A kit and method for joint detection of EPSPS and BT proteins |
| DE602005016286D1 (en) * | 2005-12-14 | 2009-10-08 | Jordanian Pharmaceutical Mfg | Apparatus for early and rapid immunochromatographic detection of HIV and its use |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016035099A1 (en) * | 2014-09-05 | 2016-03-10 | Meril Diagnostics Private Limited | A flow through device for detection of multiple bioanalytes and a process thereof |
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| WO2008104081A1 (en) | 2008-09-04 |
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