IPC분류정보
국가/구분 |
United States(US) Patent
등록
|
국제특허분류(IPC7판) |
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출원번호 |
US-0731136
(2010-03-24)
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등록번호 |
US-8415171
(2013-04-09)
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발명자
/ 주소 |
- Rissin, David M.
- Fournier, David
- Duffy, David C.
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출원인 / 주소 |
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대리인 / 주소 |
Wolf, Greenfield & Sacks, P.C.
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인용정보 |
피인용 횟수 :
12 인용 특허 :
93 |
초록
▼
Described herein are systems and methods for extending the dynamic range of assay methods and systems used for determining the concentration of analyte molecules or particles in a fluid sample. In some embodiments, a method comprises spatially segregating a plurality of analyte molecules in a fluid
Described herein are systems and methods for extending the dynamic range of assay methods and systems used for determining the concentration of analyte molecules or particles in a fluid sample. In some embodiments, a method comprises spatially segregating a plurality of analyte molecules in a fluid sample into a plurality of locations. At least a portion of the locations may be addressed to determine the percentage of said locations containing at least one analyte molecule. Based at least in part on the percentage, a measure of the concentration of analyte molecules in the fluid sample may be determined using an analog, intensity-based detection/analysis method/system and/or a digital detection/analysis method/system. In some cases, the assay may comprise the use of a plurality of capture objects.
대표청구항
▼
1. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising: exposing a plurality of capture objects, each including a binding surface having affinity for at least one type of analyte molecule or particle, to a solution containing or sus
1. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising: exposing a plurality of capture objects, each including a binding surface having affinity for at least one type of analyte molecule or particle, to a solution containing or suspected of containing the at least one type of analyte molecules or particles, wherein at least some of the capture objects become associated with at least one analyte molecule or particle;spatially segregating at least a portion of the capture objects subjected to the exposing step into a plurality of locations;addressing at least some of the plurality of locations and determining a measure indicative of the percentage of said locations containing a capture object associated with at least one analyte molecule or particle, wherein the locations addressed are locations which contain at least one capture object; andbased upon the percentage, either determining a measure of the concentration of analyte molecules or particles in the fluid sample based at least in part on the number of locations containing a capture object associated with at least one analyte molecule or particle, or determining a measure of the concentration of analyte molecules or particles in the fluid sample based at least in part on a measured intensity level of a signal of that is indicative of the presence of a plurality of analyte molecules or particles. 2. The method of claim 1, wherein the plurality of capture objects comprise a plurality of beads. 3. The method of claim 1, wherein when the percentage of said locations containing at least one capture object which contain a capture object associated with at least one analyte molecule or particle is less than about 40% the measure of the concentration of analyte molecules or particles in the fluid sample is based at least in part on the number of locations containing at least one analyte molecule or particle. 4. The method of claim 1, wherein when the percentage of said locations containing at least one capture object which contain a capture object associated with at least one analyte molecule or particle is greater than about 30% the measure of the concentration of analyte molecules or particles in the fluid sample is based at least in part on an intensity level of the at least one signal indicative of the presence of a plurality of analyte molecules or particles. 5. The method of claim 1, wherein when the percentage of said locations containing at least one capture object which contain a capture object associated with at least one analyte molecule or particle is between about 30% and about 50% the measure of the concentration of analyte molecules or particles in the fluid sample is an average, and the measure of the concentration of analyte molecules or particles in the fluid sample is based at least in part on the number of locations containing at least one analyte molecule or particle and the measure of the concentration of analyte molecules or particles in the fluid sample is based at least in part on an intensity level of the at least one signal indicative of the presence of a plurality of analyte molecules or particles. 6. The method of claim 1, further comprising at least one background signal determination. 7. The method of claim 1, wherein the measure of the concentration of analyte molecules in a fluid sample is based at least in part on comparison of a measured parameter to a calibration curve. 8. The method of claim 1, wherein the calibration curve is formed at least in part by determination of at least one calibration factor. 9. The method of claim 8, wherein the calibration factor is determined using a calibration sample wherein the percentage of locations containing or capture objects associated with at least one analyte molecule is between about 30% and about 50%. 10. The method of claim 1, wherein the measure of the concentration of analyte molecules in a fluid sample is based at least in part on a Poisson distribution adjustment. 11. The method of claim 1, wherein the plurality of capture objects comprises a plurality of beads. 12. The method of claim 1, wherein the number of said locations containing at least one analyte molecule is determined using optical techniques. 13. The method of claim 12, wherein the optical techniques comprise the use of a CCD detector. 14. The method of claim 1, wherein the plurality of locations comprises a plurality of reaction vessels. 15. The method of claim 14, wherein the plurality of reaction vessels is formed on the end of a fiber optic bundle. 16. The method of claim 14, further comprising sealing the plurality of reaction vessels. 17. The method of claim 14, wherein the average volume of the plurality of reaction vessels is between about 10 attoliters and about 100 picoliters. 18. The method of claim 1, wherein the concentration of analyte molecules or particles in the fluid sample is less than about 50×10−15 M. 19. The method of claim 1, wherein the percentage of locations addressed in the addressing step is at least about 5%. 20. The method of claim 1, wherein the analyte molecules or particles are proteins or nucleic acids. 21. The method of claim 1, further comprising performing at least one wash step. 22. The method of claim 1, wherein the analyte molecules or particles comprise an enzymatic component. 23. The method of claim 1, wherein the analyte molecules are exposed to at least one binding ligand under conditions such that substantially all of the analyte molecules associate with at least one binding ligand. 24. The method of claim 23, wherein the at least one binding ligand comprises an enzymatic component. 25. The method of claim 23, further comprising exposing the analyte molecules or particles to a precursor labeling agent. 26. The method of claim 25, wherein the precursor labeling agent is converted to a labeling agent upon exposure to the enzymatic component. 27. The method of claim 26, wherein the presence of an analyte molecule or particle at a location is determined by determining the presence of at least one precursor labeling agent.
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