Circulating microRNA as a marker for hepatocellular carcinoma
Provided herein are methods for the diagnosis, or management of liver diseases, e.g. hepatocellular carcinoma, using profiles of the miRNAs determined from cellular or acellular body fluids.
1. A detection method comprising:
(a) detecting a level of miR-16 miRNA in a serum sample from a human subject diagnosed as having liver cirrhosis by
(i) performing real-time reverse transcription polymerase chain reaction (RT-PCR) on the serum sample to measure a Ct value for miR-16 miRNA and a control Ct value for U6 small RNA, wherein the Ct value is measured using a detectably labeled probe specific for each miRNA assayed; and
(ii) calculating a delta Ct value by subtracting the U6 control Ct value from the miR-16 Ct value; and
(b) detecting at least one of the level of α-fetoprotein (AFP), the level of des-gamma-carboxyprothrombin (DCP), and the percentage of AFP-L3 in the serum sample by
(i) contacting the serum sample with an antibody that binds to AFP, an antibody that binds to DCP and/or an antibody that binds to AFP-L3; and
(ii) detecting the binding between AFP and the antibody that binds to AFP, the binding between DCP and the antibody that binds to DCP and/or the binding between AFP-L3 and the antibody that binds to AFP-L3.
2. The method of claim 1 , wherein the level of miR-16, AFP, and DCP, and the percentage of AFP-L3 are detected in the sample.
3. The method of claim 1 , wherein the level of AFP, the percentage AFP-L3, and the level of des-gamma-carboxyprothrombin (DCP) in the serum sample is greater than a reference value.
4. The method of claim 3 , wherein the reference value for: AFP is 20 ng/mL, percentage AFP-L3 is 10%, and DCP is 7.5 ng/ml.
5. The method of claim 1 , wherein the delta Ct value is greater than 6.