Methods and kits for miRNA isolation and quantitation
View Patent ↗The present invention is a kit and method for isolating and quantitating miRNA and to the use of such methods in the diagnosis and prognosis of disease.
1. A kit for isolating and analyzing miRNA from a biological fluid comprising
(a) a denaturing solution containing 20-25 mM sodium citrate and glycogen;
(b) extraction reagents;
(c) reverse transcriptase (RT) and an RT primer of SEQ ID NO:1;
(d) reagents for real time PCR; and
(e) mature miRNA.
2. The kit of claim 1 , wherein the mature miRNA is mature miR-1.
3. A method for diagnosing or prognosing an acute myocardial infarction comprising
(a) providing a blood or plasma sample from a subject;
(b) quantitating the amount of miR-1 in the subject's sample using a primer of SEQ ID NO:1; and
c) comparing the amount of miR-1 in the subject's sample with the amount of miR-1 in a reference sample, wherein an increase in the amount of miR-1 in the subject's sample compared with the reference sample is indicative of an acute myocardial infarction diagnosis or prognosis.
4. The method of claim 3 , wherein the amount of miR-1 is indicative of the amount of cardiac damage.
5. The method of claim 3 , wherein the amount of miR-1 is determined within 24 hours of the myocardial infarction.