Solid chip constant temperature detection method of MiRNA

• Main Authors: FU WEILING, XIA HAN, HUANG JUNFU, HUANG QING
• Format: Patent
• Language: English
• Published: 18.11.2015
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a solid chip constant temperature detection method of MiRNA. For an upstream primer and a downstream primer with target molecule specificity, anchoring sequence areas contain nicking agent recognition sequence positive-sense strand sequences, recognition sequence areas are complementary to the 3'-end sequences of the miRNA and the complementary chain thereof, 'cutting-extending-strand displacement' constant temperature linear amplifications which are independent and have interaction are mediated under the synergistic effect of a nicking agent and DNA polymer, and finally exponential amplification of miRNA is realized under steady temperature condition; and by fixing the upstream primer and/or the downstream primer labeled by fluorescence reporter molecules and cancellation molecules on a solid chip, the high throughput detection on various target molecules can be realized by detecting fluorescence signals. With the method, the design difficulty of the amplification detection method caused by the inherent attributes of miRNA is overcome, a liquid constant temperature index amplification and detection system based on the oligonucleotide primer is provided, and constant temperature index amplification and detection of a plurality of target molecules can be realized under the steady temperature condition.