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Nucleic acid aptamer capable of detecting myohemoglobin, microfluidic chip for screening and screening method and application

A microfluidic chip and nucleic acid aptamer technology, applied in the field of protein detection, achieves the effect of short sequence length, good stability, and reduced screening process

Inactive Publication Date: 2014-02-26
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, current antibody-based detection techniques still have certain limitations

Method used

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  • Nucleic acid aptamer capable of detecting myohemoglobin, microfluidic chip for screening and screening method and application
  • Nucleic acid aptamer capable of detecting myohemoglobin, microfluidic chip for screening and screening method and application
  • Nucleic acid aptamer capable of detecting myohemoglobin, microfluidic chip for screening and screening method and application

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Embodiment

[0051] A nucleic acid aptamer of the present invention that can be used to detect myoglobin, the nucleotide sequence of the nucleic acid aptamer includes a DNA fragment shown in any one of the following sequences 1 to 5, and sequences 1 to 5 are respectively Named Myo40-7-23, Myo40-7-27, Myo40-7-34, Myo40-7-69 and Myo19-7-34:

[0052] Sequence 1 Myo40-7-23:

[0053] 5'-GACATCTATCGCCTCGATTTCCTTTGGTGAGATCGGCTCC-3';

[0054] Sequence 2 Myo40-7-27:

[0055] 5'-CCCTCCTTTCCTTCGACGTAGATCTGCTGCGTTGTTCCGA-3';

[0056] Sequence 3 Myo40-7-34:

[0057] 5'-ACGCACAATTCCTTGTCCAATTAGGAAATTCTACGCGGAT-3';

[0058] Sequence 4 Myo40-7-69:

[0059] 5'-CGAGTACTTCTTTGCTAGTTCGCGAGATACGTTGGCTAGG-3';

[0060] Sequence 5 Myo19-7-34:

[0061] 5'-TTCCTTGTCCAATTAGGAA-3'.

[0062] In the nucleic acid aptamer that can be used to detect myoglobin in this embodiment, a certain position on its nucleotide sequence can be phosphorylated, methylated, aminated, sulfhydrylated or isotoped, and its nucleotide...

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Abstract

The invention discloses a nucleic acid aptamer capable of detecting myohemoglobin and a derivative of the nucleic acid aptamer. The nucleotide sequence of the nucleic acid aptamer comprises a DNA segment shown by any one sequence in the sequences from sequence 1-sequence 5 shown in the description. The invention discloses a microfluidic chip which comprises a sample inlet, a sample outlet and a sample feeding channel. The sample feeding channel is divided by two narrow channels into front, middle and rear channels. The front channel is filled with reverse screening protein modified microspheres, and the middle channel is filled with positive screening protein modified microspheres, wherein the grain size of microspheres is greater than width of the narrow channels. The invention further discloses a screening method for screening the nucleic acid aptamer by using the microfluidic chip. The method comprises the main steps of optimizing a nucleic acid library, screening initially, purifying, and circulating. The detection method provided by the invention has the advantages of high affinity and specificity.

Description

technical field [0001] The invention relates to the technical field of protein detection, in particular to a nucleic acid aptamer for protein detection, a screening method and application of the nucleic acid aptamer. Background technique [0002] Myoglobin (Myo) is an oxygen-collecting protein present in mammalian muscle red tissue. It consists of 153 amino acid residues, contains heme, and is homologous to hemoglobin. Under normal circumstances, it exists in the serum of healthy people in a trace form (male: 20-80 μg / L; female: 10-70 μg / L). When muscles are damaged, a large amount of myoglobin will enter the blood, causing its content in the blood to rise sharply. Myoglobin has been used as a marker for early diagnosis of myocardial infarction. In recent years, the relationship between myoglobin and myocardial infarction has been paid more and more attention. At present, the most conventional method for detecting myoglobin is to use the antigen-antibody reaction of myogl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115C12N15/10C12M1/00G01N33/68
Inventor 羊小海王青王柯敏邢煜骞江锐
Owner HUNAN UNIV
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