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Probes, kits and methods for detecting miRNAs based on cascade amplification of coded suspension microchips

A detection kit and microchip technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as limiting wide application, avoid spectral cross-interference, strong specificity and specificity. Good results

Active Publication Date: 2021-10-26
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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Problems solved by technology

Conventional nucleic acid amplification techniques often require complex and sophisticated sequence design to meet the requirements of multiple analysis of miRNAs, which also limits their wide application in the flux analysis of miRNAs.

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  • Probes, kits and methods for detecting miRNAs based on cascade amplification of coded suspension microchips
  • Probes, kits and methods for detecting miRNAs based on cascade amplification of coded suspension microchips
  • Probes, kits and methods for detecting miRNAs based on cascade amplification of coded suspension microchips

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Embodiment Construction

[0026] In view of the deficiencies in the prior art, the inventor of this case was able to propose the technical solution of the present invention after long-term research and extensive practice. The technical solution, its implementation process and principle will be further explained as follows.

[0027] As mentioned above, in view of the many defects in the prior art, the inventor of this case has been able to propose the technical solution of the present invention after long-term research and extensive practice, which is mainly through the isothermal cascade amplification based on rolling circle amplification (RCA) The technology is combined with the coded suspension microchip technology to provide a highly sensitive and specific method for the analysis of multi-component miRNAs.

[0028] Further, a method for detecting miRNAs based on cascade amplification of coded suspension microchips provided in the embodiments of the present invention may include isothermal cascade am...

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Abstract

The invention discloses a probe, a kit and a method for detecting miRNAs based on the cascade amplification of coded suspension microchips. The method for cascade amplification detection of miRNAs based on coded suspension microchips comprises: using target miRNAs as a template to perform a ligation reaction with padlock probes and RNA ligase in a ligation buffer to realize circularization of the padlock probes; Mixing the circular padlock probes with rolling circle amplification primers, polymerase, endonuclease and other components required for isothermal cascade amplification to carry out isothermal cascade amplification reaction; The product of the amplification reaction is combined with the suspension coding microchip and the universal label coupled with the capture probe to perform a base stacking hybridization reaction in a hybridization buffer; and after the base stacking hybridization reaction is completed, an optical detection device The above-mentioned suspended coding microchip is observed to realize the detection of target miRNAs. The invention can realize low-cost, high-sensitivity and high-throughput detection of tested samples.

Description

technical field [0001] The invention relates to a method for detecting microRNAs (miRNAs), in particular to a method, probe, kit, system, etc. for detecting microRNAs based on cascade amplification of coded suspension microchips. Background technique [0002] MicroRNAs (miRNAs) are small endogenous non-coding RNAs that can bind to the 3' untranscribed region (3'UTR) of messenger RNA (messengerRNA, mRNA) and inversely regulate gene expression levels at the post-transcriptional stage . Current research shows that miRNAs form a complex and highly ordered regulatory network in organisms, and almost participate in every process of normal life activities. Therefore, the analysis of miRNAs can not only be used to understand its regulatory network, but also has important value and significance in clinical application. However, miRNAs have short fragments (only 19-25 bases), low abundance in cells, and very highly homologous similar sequences. These characteristics have brought ser...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12Q1/6844
CPCC12N15/113C12N2310/10C12Q1/6844C12Q2521/301C12Q2563/107C12Q2531/125
Inventor 李炯刘胜权房涵王娜娜
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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