Method for rapidly assaying tumor-associated small peptide MUC1

A tumor-related and rapid technology, applied in the field of fluorescent probe detection, can solve the problems of time-consuming and laborious, and achieve the effect of simple operation and rapid response

Inactive Publication Date: 2016-07-20
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current methods for detecting small peptides, such as enzyme-linked immunosorbent assay, label-free surface plasmon resonance, liquid chromatography-mass spectrometry, and electrochemical analysis, although they have high sensitivity and specificity, all require a series of Fixation, separation and washing steps, time consuming and laborious

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  • Method for rapidly assaying tumor-associated small peptide MUC1

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Embodiment 1

[0023] A method for rapidly detecting tumor-associated small peptide MUC1, characterized in that it comprises the following steps:

[0024] (1) Probe pre-denaturation

[0025] First, carboxyfluorescein FAM-labeled hairpin probe 1, hairpin probe 2 and aptamer, the English name is aptamer, were incubated at 95oC for 0.5h; these three solutions were placed at 25oC for 1h;

[0026] (2) Mixed reaction

[0027] The volume of the total reaction mixture in the following steps is 200 μL, and different concentrations of mucin MUC1, hairpin probe 1 with a concentration of 10 nM, hairpin probe 2 with a concentration of 10 nM and aptamer with a concentration of 10 nM are mixed in the buffer solution, The mixed solution was reacted at 25oC for 0.5h;

[0028] The composition of the buffer solution is sodium chloride with a concentration of 0.75M and disodium hydrogen phosphate of 50mM, and the pH value is 7.4;

[0029] (3) Background quenching

[0030] Add a graphene oxide solution with ...

Embodiment 2

[0034] The detection steps are the same as Example 1, except that in step 2, the concentration of the hairpin probe 1 is 50 nM, the concentration of the hairpin probe 2 is 50 nM, and the sequence is that the reaction time is 1 h.

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Abstract

The invention discloses a method for rapidly assaying tumor-associated small peptide MUC1, which relates to the technical field of fluorescent probe assay. By utilizing the characteristic of specific recognition between the aptamer and the target small peptide, a special aptamer sequence is designed, and thereby the purpose of specific recognition of target objects is achieved; a signal amplification strategy is carried out in combination with the hybridization reaction of two types of hairpin probes, and thereby the sensitive assay of the target small peptide is realized. The method includes four steps, i.e. probe pre-denaturation, mixing reaction, background quenching and fluorescent assay, operation is easy and fast, time and labor can be saved, and a novel method is provided for the assay of tumor-associated small peptide MUC1.

Description

technical field [0001] The invention relates to the technical field of fluorescent probe detection, and more specifically relates to a small peptide fluorescent detection method constructed in the manner of specific recognition of aptamers and amplification of hybridization reactions of two hairpin probes. Background technique [0002] In recent years, the national cancer morbidity and mortality have shown a trend of increasing gradually. Therefore, it is very important to find a rapid, quantitative and sensitive method for detecting tumor markers in early detection and treatment monitoring. Currently, cancer-related tumor markers mainly include enzymes, hormones, antigens, small peptides, and small RNAs. As a product of protein hydrolysis in the human body, small peptides play an important role in the digestion and absorption of amino acids. When human cells become cancerous, the metabolism of cancer cells will be very different from that of normal cells, and cancer cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 于京华马超刘海云颜梅葛慎光张彦李丽田甜
Owner UNIV OF JINAN
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