RNA isolation from soluble urine fractions
AssigneeQuest Diagnostics Investments Incorporated
InventorsHeather Sanders, Hai-Rong Li
Life of this patent
Estimating the term…
Abstract
The invention provides methods for isolating RNA from the soluble fraction of urine. The methods can be used for detecting the presence or absence of an RNA, or quantifying the amount of an RNA. The methods are useful for diagnosing an individual suspected of having a disease by detecting the level of RNA associated with the disease in the soluble fraction of urine. The methods are also useful for prognosing an individual diagnosed with a disease by detecting the level of RNA associated with the disease in the soluble fraction of urine.
Claims24
A method for processing urine for detection of RNA expression associated with a benign prostate hyperplasia, comprising:
- a) separating urine sediment from the soluble urine fraction of a urine sample obtained from an individual suspected of having cancer, wherein RNA associated with the benign prostate hyperplasia is present in the soluble urine fraction;
- and
- b) concentrating the soluble urine fraction by ultrafiltration to produce a soluble urine concentrate, wherein the volume of the soluble urine concentrate is reduced at least 50% from the original urine volume,
- wherein the benign prostate hyperplasia-associated RNA is selected from the group consisting of HSPD1, IMPDH2, PDLIM5, and UAP1.
A method for amplifying nucleic acids from a urine sample for detection of RNA expression associated with a benign prostate hyperplasia, comprising;
- a) separating urine sediment from the soluble urine fraction of a urine sample obtained from an individual suspected of having cancer, wherein RNA associated with the benign prostate hyperplasia is present in the soluble urine fraction,
- b) concentrating the soluble urine fraction by ultrafiltration to produce a soluble urine concentrate, wherein the volume of the soluble urine concentrate is reduced at least 50% from the original urine volume,
- c) isolating RNA from the soluble urine concentrate produced in step (b),
- d) reverse transcribing the RNA from the soluble urine concentrate produced in step (c) to form cDNA;
- and
- e) amplifying the cDNA produced in step (d),
- wherein the benign prostate hyperplasia-associated RNA is selected from the group consisting of HSPD1, IMPDH2, PDLIM5, and UAP1.
A method for isolating nucleic acids from a urine sample for detection of RNA expression associated with a benign prostate hyperplasia, said method comprising,
- a) providing a urine sample from an individual suspected of having a cancer,
- b) processing said urine sample to produce a cell-free urine sample, wherein RNA associated with the benign prostate hyperplasia is present in the cell-free urine sample, and
- c) subjecting said cell-free urine sample to ultrafiltration using a filter having a nominal molecular weight limit of between about 3 kDa and about 100 kDa, to produce a soluble urine concentrate, wherein the RNA associated with the cancer is retained in the soluble urine concentrate, and wherein the volume of the soluble urine concentrate is reduced at least 50% from the original urine volume,
- wherein the benign prostate hyperplasia-associated RNA is selected from the group consisting of HSPD1, IMPDH2, PDLIM5, and UAP1.
A method for processing urine for detection of RNA expression associated with benign prostate hyperplasia comprising concentrating a urine sample from an individual suspected of having benign prostate hyperplasia to produce a soluble urine concentrate wherein RNA associated with benign prostate hyperplasia present in the urine sample is retained in the soluble urine concentrate, wherein the volume of the soluble urine concentrate is reduced at least 50% from the original urine volume, and wherein the benign prostate hyperplasia-associated RNA is selected from the group consisting of HSPD1, IMPDH2, PDLIM5, and UAP1.
Dependencies are inferred from the claim text.