Rapid and sensitive detection of cells and viruses
First Claim
1. A method for detecting individual target cells or viruses in a liquid sample, wherein said sample is added to a container comprising a detection surface and said targets measure less than 50 microns in at least two orthogonal dimensions,wherein said method comprises the steps of:
- a. contacting said individual targets with one or more signaling moieties that specifically bind to the targets,b. depositing said targets on said detection surface so that they are randomly dispersed in a detection zone comprising a detection area at a density of less than 100 target cells per mm2,c. illuminating said individual targets to generate a detectable signal from said one or more signaling moieties, andd. simultaneously detecting said individual targets by detecting said signal in a section of said detection area using a photoelectric detector, wherein the longest linear dimension of said section is greater than 1 mm, using magnification of less than or equal to 5 times, wherein said one or more signaling moieties that do not bind to said targets are not removed from said container prior to said detecting.
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Abstract
The invention provides efficient methods for rapidly and sensitively identifying cellular and viral targets in medical, industrial, and environmental samples. The invention labels targets and then detects them using large area imaging. Diagnostic tests based on the invention can be rapid, ultrasensitive, quantitative, multiplexed, and automated. The tests minimize sample preparation and do not require nucleic acid amplification or cell culture. A broad range of cells and viruses can be detected by the tests. Tests based on the invention can deliver the high level sensitivity of nucleic acid amplification tests, the user-friendliness, and speed of immunoassays, as well as the cost effectiveness and quantification offered by microbiological tests. The invention embodies the best attributes of the current diagnostic technologies, while addressing gaps in the diagnostic repertoire.
174 Citations
96 Claims
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1. A method for detecting individual target cells or viruses in a liquid sample, wherein said sample is added to a container comprising a detection surface and said targets measure less than 50 microns in at least two orthogonal dimensions,
wherein said method comprises the steps of: -
a. contacting said individual targets with one or more signaling moieties that specifically bind to the targets, b. depositing said targets on said detection surface so that they are randomly dispersed in a detection zone comprising a detection area at a density of less than 100 target cells per mm2, c. illuminating said individual targets to generate a detectable signal from said one or more signaling moieties, and d. simultaneously detecting said individual targets by detecting said signal in a section of said detection area using a photoelectric detector, wherein the longest linear dimension of said section is greater than 1 mm, using magnification of less than or equal to 5 times, wherein said one or more signaling moieties that do not bind to said targets are not removed from said container prior to said detecting. - 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, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96)
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Specification