Human cardiac troponin I aptamer, screening method and application

A cardiac troponin, aptamer technology, applied in animal/human peptides, DNA preparation, DNA/RNA fragments, etc., can solve the problems of waste of manpower, material resources and time, lack of high affinity, and few rounds. , to increase the number of washes, reduce non-specific reactions, and achieve the effect of high affinity

Inactive Publication Date: 2013-11-06
TIANJIN UNIV OF SCI & TECH
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  • Abstract
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Problems solved by technology

[0004] At present, SELEX technology is a new combinatorial chemical technology. Although it can also be used to screen for cardiac troponin I aptamers, the current general SELEX screening cycle is 8-15 rounds, and the number of rounds is too small to obtain aptamers. If there are no high-affinity aptamers, too many rounds will waste manpower, material resources and time

Method used

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  • Human cardiac troponin I aptamer, screening method and application
  • Human cardiac troponin I aptamer, screening method and application
  • Human cardiac troponin I aptamer, screening method and application

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Embodiment Construction

[0059] The present invention will be further described in detail below through the specific examples, the following examples are only descriptive, not restrictive, and cannot limit the protection scope of the present invention with this.

[0060] The various substances used in the preparation method of the present invention are commercially available products unless otherwise specified; the methods used are special methods unless otherwise specified.

[0061] In this experiment, SELEX technology was used to screen cardiac troponin I protein aptamers using the microwell plate method. First, the randomly synthesized single-stranded oligonucleotide library was combined with the protein, and the unbound oligonucleotide library was washed. The bound sequence is eluted and used as a template for PCR amplification. Secondary libraries are then prepared by asymmetric PCR, followed by binding to the target protein, and repeated for multiple cycles. Through repeated screening and ampli...

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Abstract

The invention relates to a human cardiac troponin I aptamer which has a nucleotide sequence in a sequence table SEQ ID No.1 or a nucleotide sequence in a sequence table SEQ ID No.2 or a nucleotide sequence in a sequence table SEQ ID No.3 or a nucleotide sequence in a sequence table SEQ ID No.4. The human cardiac troponin I (cardiac troponin I) aptamer is high in affinity combined with a cTn I and strong in specificity, improves cardiac troponin I application of diagnosis on early detection and clinical myocardial damage or sensitivity for separating and purifying the cardiac troponin I.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a human cardiac troponin I aptamer, in particular to a human cardiac troponin I aptamer, a screening method and application. Background technique [0002] Acute myocardial infarction (AMI) is one of the cardiovascular diseases with a high mortality rate in my country, and its early diagnosis and timely treatment are the key to reducing the mortality rate. Cardiac troponin I shows its special superiority in the diagnosis of AMI and other heart diseases due to its high sensitivity, tissue and organ specificity and long diagnostic window period, and it is used as the "standard" for the detection of acute myocardial infarction. gold standard". [0003] Nucleic acid aptamers are a class of biomolecular artificial ligands that have emerged in recent years. High-affinity and specific cardiac troponin I aptamers were screened from large-capacity single-stranded oligonucleotide libraries throu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115C12N15/10C12Q1/68C07K14/47
Inventor 张娟琨刘珺蒋树海徐锦锋
Owner TIANJIN UNIV OF SCI & TECH
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