RNA sensor for detecting thiamine pyrophosphate and application thereof

Abstract

The invention discloses an RNA sensor for detecting thiamine pyrophosphate. The sensor comprises an RNA aptamer for specifically sensing thiamine pyrophosphate, linkage segment RNA and hammerhead ribozyme, wherein the RNA aptamer is connected with the hammerhead ribozyme by virtue of the linkage segment RNA; when the RNA aptamer senses the thiamine pyrophosphate, self-cleavage of the hammerhead ribozyme is triggered. The invention further discloses a kit comprising the RNA sensor disclosed by the invention and a method for detecting the thiamine pyrophosphate. The RNA sensor disclosed by the invention is high in sensitivity of detecting the thiamine pyrophosphate and capable of performing qualitative and quantitative detection on the thiamine pyrophosphate.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an RNA sensor for rapidly and sensitively detecting thiamine pyrophosphate (TPP) and an application thereof. Background technique [0002] Alzheimer's disease (AD) is a degenerative disease of the nervous system. Clinically, it is characterized by generalized dementia such as memory impairment, aphasia, agnosia, executive dysfunction, and personality and behavior changes. The most prominent pathological features of AD include deposition of β-amyloid protein in the brain, hyperphosphorylation of tau-like protein, disturbance of glucose metabolism in the brain, and loss of neurons and their synapses. TPP is the coenzyme factor of the pyruvate dehydrogenase complex in the glucose metabolism pathway, the α-ketoglutarate dehydrogenase complex and the transketolase in the pentose phosphate pathway. The lack of TPP will lead to the decrease of the activity of the above three enzymes, the di...

AI Technical Summary

Problems solved by technology

Although there are many kinds of methods used to detect TPP, all of them have certain limitations and shortcomings.

For example, the detection method based on liquid chromatography-mass spectrometry, although the detection results of this technology are more accurate and reproducible, but it requires professional operation, and the equipment is sophisticated and expensive, and most importantly, it cannot achieve clinical high-throughput detection

Although intracellular fluorescent biosensors use the principle of riboswitches and have good specificity, they require high and cumbersome operations. In addition, this method requires a high level of background concentration (micromol / L, umol / L). Therefore, this method is not suitable for the detection of TPP content in human biological samples (such as in blood: nmol / liter, nmol / L)

Nanogold-based detection methods can achieve lower detection limits, but the specificity of this detection method is not high, such as binding other small molecules through electrostatic interactions, resulting in false positive signals

Although the detection methods based on electrochemical method and chemiluminescence method are relatively convenient and fast, they all have the disadvantage of low specificity for TPP recognition.

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