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Biosensor based on bismuth-olefin nanosheet fluorescence quenching, miRNA detection kit and application of miRNA detection kit

A technology of biosensors and detection kits, applied in the biomedical field, can solve the problems of large demand for RNA samples, inability to distinguish miRNA, and low detection resolution, and achieve small sample demand, fast detection speed, and high detection specificity sexual effect

Active Publication Date: 2020-04-10
SHENZHEN HANGUANG TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In the first aspect, the present invention provides a biosensor based on fluorescence quenching of bismuthene nanosheets to solve the problems of cumbersome operation, large demand for RNA samples, inability to distinguish miRNAs with small differences, and detection low resolution issues

Method used

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  • Biosensor based on bismuth-olefin nanosheet fluorescence quenching, miRNA detection kit and application of miRNA detection kit
  • Biosensor based on bismuth-olefin nanosheet fluorescence quenching, miRNA detection kit and application of miRNA detection kit
  • Biosensor based on bismuth-olefin nanosheet fluorescence quenching, miRNA detection kit and application of miRNA detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1: Biosensor based on bismuthene nanosheet fluorescence quenching

[0042] A biosensor based on the fluorescence quenching of bismuthene nanosheets, including fluorescently labeled single-stranded nucleic acid probes and bismuthene nanosheets. The single-stranded nucleic acid probes are adsorbed on the surface of bismuthene nanosheets by van der Waals force to form a unique fluorescence A complex structure of the element-single-stranded nucleic acid probe-bismuthene nanosheet, wherein the single-stranded nucleic acid probe is complementary to a specific miRNA, and is used to combine with the specific miRNA to form a double-stranded structure.

[0043] The detection principle is: by virtue of the unique fluorescence quenching effect of bismuthene nanosheets, the fluorescence of the fluorescein-single-stranded nucleic acid probe-bismuthene nanosheets complex is quenched. The remarkable quenching effect of bismuth ascribes the weak ground-state fluorescent charge t...

Embodiment 2

[0054] Embodiment 2: miRNA detection kit

[0055] A miRNA detection kit includes two parts: a fluorescently labeled single-stranded nucleic acid probe and a fluorescent quencher, wherein the single-stranded nucleic acid probe is complementary to a specific miRNA, and the fluorescent quencher includes bismuthene nanosheets. For example, the miRNA detection kit may include two parts of reagents, the first part of reagents includes a certain concentration of fluorescently labeled single-stranded nucleic acid probes, and the second part of reagents includes a certain concentration of fluorescence quenchers.

[0056] When using the miRNA detection kit, the fluorescently labeled single-stranded nucleic acid probe can be first incubated with a fluorescent quencher to form a fluorescein-single-stranded nucleic acid probe-bismuthene nanosheet complex, that is, based on the bismuthene nanosheet fluorescence In the quenched biosensor, at this time, the fluorescence of the fluorescein-sin...

Embodiment 3

[0061] Example 3: Preparation and Characterization of Bismuthene Nanosheets

[0062] In this example, commercially available bulk bismuth was treated by sonochemical method, few-layer bismuthene nanosheets were prepared by top-down liquid phase exfoliation method, and the morphology and chemical composition of the prepared bismuthene nanosheets were characterized.

[0063] (1) Preparation of bismuthene nanosheets: liquid-phase ultrasonic treatment is an effective method to break the weak van der Waals force to prepare multilayer bismuth.

[0064] Such as figure 1 As shown in a, well-dispersed and ultrathin bismuth nanosheets were successfully prepared using the probe ultrasonic liquid phase exfoliation method. Specifically, 30 mg of bismuth powder was ground and added to a solvent (300 ml). The bismuth powder solution was sonicated at 420 W for 6 h in an ice bath. Then, the solution was ultrasonically treated by a 1080W probe for 24h. Subsequently, the solution was centrif...

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Abstract

The invention provides a biosensor based on bismuth-olefin nanosheet fluorescence quenching. The biosensor comprises a fluorescence-labeled single-stranded nucleic acid probe and a bismuth-olefin nanosheet, wherein the single-stranded nucleic acid probe is complementary to a specific miRNA; the single-stranded nucleic acid probe is adsorbed on the surface of the bismuth-olefin nanosheet; when thesingle-stranded nucleic acid probe is adsorbed on the surface of the bismuth-olefin nanosheet, fluorescence is quenched; and when the specific miRNA competitively binds with the single-stranded nucleic acid probe, the fluorescence is recovered. The biosensor based on bismuth-olefin nanosheet fluorescence quenching has the advantages of extremely high detection limit, extremely high detection specificity, simple operation, high detection speed, small sample demand and the like when being used for detecting the specific miRNA. The invention also provides a miRNA detection kit and an applicationthereof.

Description

technical field [0001] The present invention relates to the technical fields of biomedicine and analytical chemistry, in particular to a reagent for clinical diagnosis, more specifically to a biosensor based on bismuthene nanosheet fluorescence quenching, and including the fluorescence quenching sensor based on bismuthene nanosheet The miRNA detection kit of the biosensor, and the present invention also relates to the application of the biosensor based on bismuthene nanosheet fluorescence quenching in the detection of miRNA. Background technique [0002] MicroRNA (miRNA) is a single-stranded small molecule RNA expressed endogenously in the body. It is located in the non-coding region of the genome and does not have an open reading frame (open reading frame, ORF). . miRNA widely exists in various eukaryotic cells, does not encode any protein, and is only 20-24nt in length. There is a phosphate group at the 5' end of the mature miRNA, and a hydroxyl group at the 3' end, whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6883C12Q1/6876
CPCC12Q1/6886C12Q1/6883C12Q1/6876C12Q2600/178Y02A50/30
Inventor 梁维源张家宜范涛健康建龙黄浩
Owner SHENZHEN HANGUANG TECH CO LTD
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