miRNA fingerprint in the diagnosis of diseases

摘要

The present invention provides novel methods for diagnosing a state of health based on the determination of specific miRNAs that have altered expression levels in different conditions, e.g. disease states compared to healthy controls.

主权项

1. A method of diagnosing melanoma, comprising the steps
(a) determining an expression profile of a predetermined set of miRNAs in a biological sample from a patient, wherein said biological sample is a blood cell sample by:
(i) providing capture probes, each capture probe comprising a sequence of at least partially complementary nucleotides to each of the miRNAs of the predetermined set of miRNAs and an elongation sequence, and(ii) hybridizing the miRNAs of said biological sample of (a) to the capture probes, and(iii) elongating the hybridized miRNA of step (ii) with at least one labeled nucleotide complementary to the elongation sequence, and(iv) quantifying the labeled hybridized miRNAs based on the labels or(v) obtaining cDNA samples from the biological sample of (a) by a RNA reverse transcription reaction using miRNA-specific primers, and(vi) amplifying the cDNA via polymerase chain reaction (PCR) with miRNA-specific cDNA forward primers, universal reverse primers and a labeled probe complementary to at least a portion of the cDNA samples to be analyzed, and(vii) detecting the levels of miRNAs on the basis of the labeled probe, (b) comparing said expression profile to a reference expression profile, wherein the comparison of said expression profile to said reference expression profile allows for the diagnosis of melanoma, and
wherein the predetermined set of miRNAs representative for melanoma comprises at least 1, 3, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 miRNAs selected from the group consisting of: (i) the miRNAs from FIG. 30A (ii) the miRNAs from FIG. 30B (iii) the miRNAs from FIG. 32A (iv) the miRNAs from FIG. 32B (v) the miRNAs from the signatures in FIG. 33 A and (vi) the miRNAs from the signatures in FIG. 33B.