Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Methylene-blue-based upconversion nanoparticle-modified fibration paper sensor for detecting telomerase and preparation method thereof

A methylene blue and paper sensor technology, applied in the world, belongs to the field of biotechnology and clinical medical diagnosis, which can solve the problems of time-consuming, complicated equipment, instability, etc., and achieve the effects of short detection time, low detection cost and less reagent consumption

Inactive Publication Date: 2017-07-11
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
View PDF4 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[See: (a) Y.Xiao, K.Y.Dane, T.Uzawa, A.Csordas, J.Qian, H.T.Soh, P.S.Daugherty, E.T.Lagally, A.J.Heeger, K.W.Plaxco.J.Am.Chem.Soc.2010, 132, 15299. (b) G. Zhang, W. L. Daniel, C. A. Mirkin. J. Am. Chem. Soc. 2008, 130, 9644. (c) G. F. Zheng, F. Patolsky, Y. Cui, W. U. Wang, C. M. Lieber .Nat.Biotechnol.2005, 23, 1294.] Although these methods are ingeniously designed and somewhat make up for the shortcomings of PCR technology, most of these new methods still have a large number of detection steps, require complex instruments and equipment, and time-consuming problems. More importantly, , most of these studies focus on liquid-phase reaction systems, which are usually unstable and impractical to perform outside the laboratory
Therefore, the above assays are not suitable for point-of-care (POC) diagnosis of serious diseases

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Methylene-blue-based upconversion nanoparticle-modified fibration paper sensor for detecting telomerase and preparation method thereof
  • Methylene-blue-based upconversion nanoparticle-modified fibration paper sensor for detecting telomerase and preparation method thereof
  • Methylene-blue-based upconversion nanoparticle-modified fibration paper sensor for detecting telomerase and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Preparation of Modified Fiberized Paper Sensor

[0027] Experimental materials: ordinary fibrous filter paper, standard GB / T 1914-2007, Taizhou Jinao Filter Paper Company; anhydrous lithium chloride (LiCl), batch number F20130906, Shanghai Abby Chemical Reagent Co., Ltd.; sodium periodate (NaIO 4 ), batch number SB0875, Sangon Bioengineering (Shanghai) Co., Ltd.; the sequence of the aptamer is: TS-NH 2 : 5’-NH 2 (CH 2 ) 6 TTTTTTAATCCGTCGAGCAGAGTT-3', Sangon Bioengineering (Shanghai) Co., Ltd.

[0028] Experimental instruments and equipment: KQ-500DE ultrasonic cleaner, DHG-9143BS electric blast drying oven, AY120 electronic analytical balance, TGL-16C centrifuge.

[0029] experiment procedure:

[0030] (1) Pretreatment of fiberized paper

[0031] In a 250mL beaker, mix 0.4g LiCl and 1.0g NaIO 4 Fully dissolve in 200mL distilled water, cut the fibrous paper into 15*15mm size, soak it in a beaker, heat it in a water bath at 60°C for 60 minutes, take out ...

Embodiment 2

[0036] Embodiment 2: Telomere complementary oligonucleotide (TC-NH 2 ) modified upconverting nanoparticles

[0037] Experimental materials: bovine serum albumin, batch number 140318, Shanghai Aladdin Reagent Co., Ltd.; 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), batch number 090M14531V, Shanghai Aladdin Reagent Co., Ltd. Latin Reagent Co., Ltd.; succinimide (NHS), lot number MKBG7914V, Shanghai Aladdin Reagent Co., Ltd.; ethanol, lot number: 14021710247, purchased from Nanjing Chemical Reagent Co., Ltd. The capture aptamer sequence is: TC-NH 2 : capture aptamer (TC-NH 2 ): 5'-CCCTAACCCTAAAAAA (CH 2 ) 3 NH 2 -3', Sangon Bioengineering (Shanghai) Co., Ltd.

[0038] Experimental instruments and equipment: KQ-500DE ultrasonic cleaner, DHG-9143BS electric blast drying oven, AY120 electronic analytical balance, TGL-16C centrifuge.

[0039] Experimental process: 1 mL of carboxylated upconverting nanoparticles (UCNPs) with a concentration of 1 mg / mL was wa...

Embodiment 3

[0040] Example 3: Select human cervical cancer cells (Hela cells) as the research target, and test the fibrosis paper sensor based on methylene blue and up-conversion nanoparticles

[0041] Experimental material: ethanol, batch number: 14021710247, purchased from Nanjing Chemical Reagent Co., Ltd. HEPES buffer solution, batch number: E607018, purchased from Shanghai Sangon Bioreagent Co., Ltd.; 5′-adenosine triphosphate disodium salt trihydrate (ATP), batch number: A600020, purchased from Shanghai Sangon Bioreagent Co., Ltd.;

[0042] Experimental instruments and equipment: KQ-500DE ultrasonic cleaner, DHG-9143BS electric blast drying oven, AY120 electronic analytical balance, TGL-16C centrifuge.

[0043] experiment procedure:

[0044] (1) Extraction of human cervical cancer cell telomerase

[0045] Take the cervical cancer cells in the exponential growth phase in a 10mL centrifuge tube, wash twice with ice-cold PBS buffer, and count with a hemocytometer. μL, 80cell / μL, and...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Thicknessaaaaaaaaaa
Diameteraaaaaaaaaa
Thicknessaaaaaaaaaa
Login to View More

Abstract

The invention relates to a fibrotic paper sensor based on methylene blue and up-conversion nanoparticle modification and a preparation method thereof, which is used for detecting the activity of telomerase, a tumor marker. When the cell lysate to be tested is dropped on the modified fiber paper, the telomerase substrate can undergo different degrees of elongation reaction, and then use oligonucleotide-modified methylene blue and up-converting nanoparticles to telomere The enzyme extension product is reacted, and the detection result can be obtained by the color depth of methylene blue and the fluorescence intensity of the up-conversion nanoparticles under near-infrared irradiation. The fixed fibrous paper can better carry the detection liquid through capillary action, which is beneficial to the occurrence and progress of biological reactions. The present invention has the advantages of simple operation, low preparation cost, fast and accurate detection, strong specificity, and high sensitivity. Good application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology and clinical medical diagnosis, and relates to a preparation method of a modified fibrous paper sensor based on methylene blue and up-conversion nanoparticles, which is mainly used for the detection of tumor marker-telomerase. The capillary action carries the liquid to be tested for biological reactions, and the functionalized methylene blue and upconversion nanoparticles are used to characterize the activity of intracellular telomerase, so as to realize the early diagnosis of cancer and tumor diseases. Background technique [0002] Telomere is a special nucleic acid complex that uses its internal RNA as a template to catalyze the human chromosome 3' terminal repeat sequence (TTAGGG) through reverse transcription. n to prevent the recombination, fusion and loss of chromosomal DNA. Telomerase can regulate the length of telomeres, thereby controlling the life activities of cells. Studies have found th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/574G01N21/64
CPCG01N33/57484G01N21/6428
Inventor 傅安辰朱栋苏瑞琳
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com