Methods and device for detecting and quantifying substances in body fluids
First Claim
1. An in vivo method for determining an analyte in the body fluids of an individual comprising the steps ofa) placing a sensor in communication with the body fluids of said individual suspected of containing said analyte in such a way that once in place said sensor does not exit the skin of the individual,said sensor comprising a fluorescence reagent for detecting said analyte that reversibly binds to said analyte,said fluorescence reagent having a fluorescence intensity, an emission spectrum, an excitation spectrum, or an excited state lifetime in the presence of said analyte that is different from its fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime in the absence of said analyte,said sensor being configured to retain said fluorescence reagent while allowing said analyte to diffuse into and out of said sensor;
- b) transdermally illuminating said sensor; and
c) measuring the fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime of said fluorescence reagent relative to the fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime of said fluorescence reagent in the absence of said analyte; and
d) correlating the change in fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime of said fluorescence reagent with the presence or amount of said analyte in said individual.
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Abstract
An in vivo method and apparatus for detecting an analyte in an individual. A sensor that includes a fluorescence reagent is placed in communication with the body fluids of the individual suspected of containing the analyte in such a way that once in place said sensor does not exit the skin of the individual. The sensor is configured to retain the fluorescence reagent while allowing analyte to diffuse into and out of said sensor. The sensor is illuminated transdermally, and the fluorescence from the fluorescence reagent associated with the presence of the analyte is measured.
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Citations
30 Claims
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1. An in vivo method for determining an analyte in the body fluids of an individual comprising the steps of
a) placing a sensor in communication with the body fluids of said individual suspected of containing said analyte in such a way that once in place said sensor does not exit the skin of the individual, said sensor comprising a fluorescence reagent for detecting said analyte that reversibly binds to said analyte, said fluorescence reagent having a fluorescence intensity, an emission spectrum, an excitation spectrum, or an excited state lifetime in the presence of said analyte that is different from its fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime in the absence of said analyte, said sensor being configured to retain said fluorescence reagent while allowing said analyte to diffuse into and out of said sensor; -
b) transdermally illuminating said sensor; and c) measuring the fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime of said fluorescence reagent relative to the fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime of said fluorescence reagent in the absence of said analyte; and d) correlating the change in fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime of said fluorescence reagent with the presence or amount of said analyte in said individual. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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28. An in vivo method for determining an analyte in the body fluids of an individual comprising the steps of
a) placing a sensor in communication with the body fluids of said individual suspected of containing said analyte, said sensor comprising a fluorescence reagent for detecting said analyte that reversibly binds to said analyte, said sensor being configured to retain said fluorescence reagent while allowing analyte to diffuse into and out of said sensor, said fluorescence reagent comprising an energy-absorbing donor molecule and an energy-absorbing acceptor molecule, the excited state energy level of the donor overlapping with the excited state energy level of the acceptor; -
b) transdermally illuminating said sensor so as to i) excite the donor or ii) excite both the donor and acceptor; and c) measuring the fluorescence from said fluorescence reagent associated with the presence of said analyte in said individual by determining the extent to which non-radiative fluorescence resonance energy transfer occurs between the donor and the acceptor upon binding, said non-radiative fluorescence resonance energy transfer being determined by measuring i) the ratio of the fluorescence signal at two emission wavelengths, one of which is due to donor emission and the other of which is due to acceptor emission, when only the donor is excited, ii) the ratio of the fluorescence signal due to the acceptor following donor excitation and the fluorescence signal due to the acceptor following acceptor excitation, iii) a change in donor lifetime, iv) quenching of donor fluorescence, or v) an enhancement of acceptor fluorescence intensity; and d) correlating said non-radiative fluorescence resonance energy transfer with the presence or amount of said analyte in said individual.
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29. An in vivo sensor for determining an analyte in the body fluids of an individual comprising a fluorescence reagent for determining said analyte that reversibly binds to said analyte,
said fluorescence reagent having a fluorescence intensity, an emission spectrum, an excitation spectrum, or an excited state lifetime in the presence of said analyte that is different from its fluorescence intensity, emission spectrum, excitation spectrum, or excited state lifetime in the absence of said analyte, said sensor being configured such that (a) once implanted in the individual said sensor does not exit the skin of the individual, and (b) said fluorescence reagent is retained within said sensor while said analyte is allowed to diffuse into and out of said sensor.
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30. An in vivo sensor for determining an analyte in the body fluids of an individual comprising a fluorescence reagent for determining said analyte that reversibly binds to said analyte,
said fluorescence reagent comprising an energy-accepting donor molecule and an energy-absorbing acceptor molecule, the excited state energy level of the donor overlapping with the excited state energy level of the acceptor, said sensor being configured such that (a) once implanted in the individual said sensor does not exit the skin of the individual, and (b) said fluorescence reagent is retained within said sensor while said analyte is allowed to diffuse into and out of said sensor.
Specification