Reivindicaciones
1. A stem-loop primer comprising the nucleotide sequence SEQ ID NO: 1. 2. A composition comprising at least one stem-loop primer according to claim 1. 3. A kit for short length RNA detection comprising at least one stem-loop primer according to claim 1 or the composition according to claim 2. 4. Kit according to claim 3, further comprising a reverse transcriptase. 5. Kit according to claims 3 or 4, further comprising at least one forward primer comprising the nucleotide sequence SEQ ID NO: 49, preferably the nucleotide SEQ ID NO: 2 or SEQ ID NO: 50. 6. Kit according to any one of the claims 3 to 5, further comprising a universal reverse primer comprising the nucleotide sequence SEQ ID NO: 3 and/or a universal probe comprising the nucleotide sequence SEQ ID NO: 6, preferably the universal reverse primer comprising the nucleotide sequence SEQ ID NO: 4 or SEQ ID NO: 5 and/or the universal probe comprising the nucleotide sequence SEQ ID NO: 7. 7. An in vitro method for detecting and/or amplifying at least one target short length RNA in an isolated biological sample of a subject comprising: (i) reverse-transcribing the target short length RNA using a reverse transcriptase and the stem-loop primer according to claim 1, the composition according to claim 2 or the kit according to claim 3 or 4 obtaining a double-strand cDNA, and (ii) performing a quantitative PCR (qPCR) using as a template the double-strand cDNA obtained in (i) and - at least one forward primer comprising the nucleotide sequence SEQ ID NO: 49, preferably the nucleotide sequence SEQ ID NO: 2 or SEQ ID NO: 50; a universal reverse primer comprising the nucleotide sequence SEQ ID NO: 3, preferably the nucleotide sequence SEQ ID NO: 4 or SEQ ID NO: 5; and/or a universal probe comprising the nucleotide sequence SEQ ID NO: 6, preferably the nucleotide sequence SEQ ID NO: 7, or - the kit according to claim 5 or 6. 8. The method according to claim 7, wherein the qPCR comprises an intermediate hybridization step, preferably wherein the temperature of the hybridization step is adjusted to the average temperature of the forward primer of each targeted short length RNA. 9. The method according to claim 7 or 8, wherein the target short length RNA is a miRNA. 10. The method according to claim 9, wherein the miRNA is selected from the list consisting of: SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20 and any combination thereof. 11. The method according to any one of the claims 7 to 10, wherein the stem-loop primer sequence is selected from the list consisting of: SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 and any combination thereof. 12. The method according to any one of the claims 7 to 11, wherein the forward primer sequence is selected from the list consisting of: SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46 and any combination thereof. 13. The method according to any one of the claims 7 to 12, wherein the forward and universal reverse primer concentration is 1,5 µM and 0,7 µM respectively, the universal probe concentration is 0,8 µM and/or the stem-loop primer concentration is 1nM. 14. Use of the kit according to any of the claims 3 to 6 for in vitro short length RNA detection. 15. Use according to claim 14, wherein the short length RNA is a miRNA.
