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Biosensor for detecting tumor markers

A technology of biosensors and tumor markers, applied in the direction of biological testing, biomaterial analysis, biochemical equipment and methods, etc., can solve the problems of limiting the practical application of colorimetric pH sensing, low target abundance, etc., to overcome the low detection Abundance targets, removal of interference from complex components, effectiveness of simplified detection methods

Active Publication Date: 2020-06-12
SUZHOU CHIEN SHIUNG INST OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, low target abundance and interference from complex components and analytical procedures limit the practical application of colorimetric pH sensing

Method used

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  • Biosensor for detecting tumor markers
  • Biosensor for detecting tumor markers
  • Biosensor for detecting tumor markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation process and test method

[0039] A biosensor for detecting tumor markers is made by the following steps:

[0040] I. Use immunomagnetic beads to selectively isolate targets;

[0041] II. Bind the aptamer-trigger to the target to form a sandwich structure of immunomagnetic beads / target / aptamer-trigger;

[0042] III. The aptamer-trigger immobilized on the surface of the immunomagnetic beads initiates linear rolling circle amplification in the presence of circular template 1 to generate a product of linear rolling circle amplification;

[0043] IV. The GOD-labeled auxiliary primer 2 and the circular template 2 combine with the product of the linear rolling circle amplification in step III to form a ternary complex, and the auxiliary primer 2 starts the branched rolling circle amplification to generate branched products;

[0044] V. GOD-labeled auxiliary primer 1 and circular template 1 combine with the branched product in step IV to form another ternary compl...

experiment example

[0057] Experimental example: Feasibility verification and optimization comparison of colorimetric pH sensing.

[0058] The GOD selected by the present invention works well under neutral pH value (5.2~6.8), consistent with the range of discoloration of BCP (see appendix figure 2 A, B and attached image 3 ).

[0059] Further, neutral pH 7.0 was selected as the initial pH for further experiments, because the color change from neutral pH to acidic pH is obvious to the naked eye. attached figure 2C shows that even at a low GOD concentration (2 nM), the change from purple to yellow can be observed with the naked eye in a short time (30 min), indicating that GOD performs well in pH sensing.

[0060] attached figure 2 D provides more accurate colorimetric results. It can be seen that the A590 nm / A430 nm of all the positive samples (with GOD) had obvious changes, while the A590 nm / A430 nm of the negative samples (without GOD) had little change. The above results demonstrate t...

Embodiment 3

[0062] Experimental example: comparison of amplification efficiency of mb-RCA

[0063] as attached Figure 4 As shown, the change of pH was positively correlated with the concentration of GOD. Therefore, enriching more GOD by signal amplification technology is an effective way to improve the detection effect of low-abundance targets. In the present invention, we propose a novel isothermal DNA signal amplification technique called mb-RCA. Since mb-RCA can be divided into two closely related LRCA systems, these two LRCA systems are verified first. as attached Figure 4 As shown in A, LRCA can only be initiated in the presence of a trigger, template and polymerase. Subsequently, the DNA polymerase was optimized (attached Figure 4 B). Among the two commonly used DNA polymerases that can work normally at 37°C, the amplification efficiency of phi29-DNA polymerase is much higher than that of Klenow Fragment (3'-5'exo-) many. In addition, good amplification specificity was ob...

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Abstract

The invention discloses a biosensor for detecting tumor markers, which is based on a glucose oxidase (GOD) mediated colorimetric pH sensing sensitivity analysis technology, is used for detecting cancer biomarker proteins, and effectively integrates the following three aspects into the biosensor: magnetic separation, signal amplification and signal conversion. The magnetic separation and signal amplification process can effectively eliminate the interference of complex components and overcome the challenge of detecting low-abundance targets. Acidogenase GOD mediated signal conversion can causerapid decrease of the pH value, and preconditions are provided for a pH colorimetric method. The development based on the pH colorimetric detection technology has important value for promoting the practical application of pH sensing in POCT, and can provide guidance for the development of other high-performance sensors.

Description

technical field [0001] The invention belongs to the technical field of bioelectrochemical sensors, and in particular relates to a biosensor for detecting tumor markers and an application thereof. [0002] technical background [0003] Cancer biomarkers (such as proteins, nucleic acids, small molecules, exosomes, and ring tumor cells) are important detection indicators for early cancer screening and diagnosis. Over the past few decades, researchers have developed a variety of techniques for the rapid and sensitive detection of biomarkers. However, in practical applications, accepted detection techniques are still traditional techniques. For example, in the detection of cancer biomarker proteins, today's main detection technology is still based on enzyme-linked immunosorbent assay (ELISA) laboratory detection technology. Therefore, the optimization and innovation of detection technology still has great value in cancer diagnosis, especially the development of low-cost and conv...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574G01N33/543C12Q1/6825
CPCC12Q1/6825G01N33/54326G01N33/57484G01N33/68G01N2333/49C12Q2525/205C12Q2531/125C12Q2525/307
Inventor 苗向阳朱小立毛东升王兆寅严丹红朱志强葛林
Owner SUZHOU CHIEN SHIUNG INST OF TECH
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