Rapid bioluminescence detection system
View Patent ↗An assay is provided for detecting the activity of a reporter kinase comprising (i) adding said reporter kinase to an assay mixture wherein said reporter kinase is contacted with bioluminescent reagent no more than 5 minutes after being contacted with ADP, and wherein, prior to contacting the reporter kinase with ADP, the assay mixture is substantially free from kinase other than reporter kinase; and (ii) detecting light output from the assay mixture. Methods for detecting the presence of an analyte in a sample and methods for validating a treatment process using the above assay are also provided. Further provided are devices for conducting these assays and methods.
1. A lateral flow device for use in an assay for detecting the presence of a known analyte in a sample, comprising:
a backing strip on which is positioned an elongate flow matrix, wherein said flow matrix comprises:
(i) a sample-receiving zone containing an exogenous kinase attached to the flow matrix via linker comprising a binding agent specific for the analyte;
(ii) a detection zone, located downstream of the sample-receiving zone, and containing a mixture of ADP and a bioluminescent reagent; and
(iii) a background-reduction zone, situated between the sample-receiving zone and the detection zone, that substantially removes or inhibits kinase other than exogenous kinase present in the sample;
wherein, in use, a sample is applied to the sample-receiving zone and analyte present in the sample displaces exogenous kinase from the flow matrix, said displaced exogenous kinase migrates through the background-reduction zone where kinase other than exogenous kinase is substantially removed or inhibited, and then into the detection zone where ATP generation is detected.
2. The device according to claim 1 , wherein any ATP present is substantially removed from the background reduction zone.
3. The device according to claim 1 , wherein the background-reduction zone comprises one or more of a substance that substantially inhibits or removes kinase other than exogenous kinase and/or ATP.
4. The device according to claim 1 , wherein the device is portable.
5. The device according to claim 3 , wherein the one or a substance that substantially inhibits or removes kinase other than exogenous kinase and/or ATP is selected from the group consisting of an immobilised ATPase, an anionic or cationic exchange matrix, a size reduction matrix and a combination thereof.
6. A lateral flow device for use in an assay for detecting the presence of a known analyte in a sample, comprising:
a backing strip on which is positioned an elongate flow matrix, wherein said flow matrix comprises:
(i) a sample-receiving zone containing a complex comprising an exogenous kinase linked to a binding agent specific for the analyte and an analyte that is bound to said matrix;
(ii) a detection zone, located downstream of the sample-receiving zone, and containing a mixture of ADP and a bioluminescent reagent; and
(iii) a background-reduction zone, situated between the sample-receiving zone and the detection zone, that substantially removes or inhibits kinase other than exogenous kinase present in the sample;
wherein, in use, a sample is applied to the sample-receiving zone and analyte present in the sample displaces exogenous kinase from the flow matrix, said displaced exogenous kinase migrates through the background-reduction zone where kinase other than exogenous kinase is substantially removed or inhibited, and then into the detection zone where ATP generation is detected.
7. The device according to claim 6 , wherein any ATP present is substantially removed from the background reduction zone.
8. The device according to claim 6 , wherein the background-reduction zone comprises one or more of a substance that substantially inhibits or removes kinase other than exogenous kinase and/or ATP.
9. The device according to claim 6 , wherein the device is portable.
10. The device according to claim 8 , wherein the one or more of a substance that substantially inhibits or removes kinase other than exogenous kinase and/or ATP is selected from the group consisting of an immobilised ATPase, an anionic or cationic exchange matrix, a size reduction matrix and a combination thereof.