Cervical cancer cell detection kit

A detection kit, cervical cancer cell technology, applied in the direction of measuring devices, microbiological determination/inspection, instruments, etc., can solve the problems of prone to photobleaching and photodegradation, poor photochemical stability, wide emission spectrum, etc., and achieve optical resistance Strong interference ability, improved detection sensitivity, and large amount of information

Active Publication Date: 2016-08-24
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional fluorescent reagents are dye molecules, such as FITC (Xiuhan Yang, Xiaochuan Wang, Xiaomin Zhang. Capillary zone electrophoresis separation of low concentration stimulants in human urine with laser-induced fluorescence detection. Analytica Chimica Acta, 2005, 549 (1-2), 81-87.), cyanine dyes Cy3, Cy5, etc., they have some unavoidable shortcomings, such as: poor photochemical stability, prone to photobleaching and photodegradation, which is not conducive to long-term observation
In addition, traditional dye molecules have narrow excitation spectra and broad emission spectra, which are easily interfered by the autofluorescence of biological macromolecules in complex life systems.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The preparation of embodiment 1 kit

[0036] (1) Preparation of B-GQDs

[0037] Boron-doped graphene quantum dots (B-GQDs) were synthesized by potentiostatic chronoamperometry via an electrochemical workstation (CHI 660B). The specific method is: prepare 0.1mol / L borax aqueous solution, and take 20mL as electrolyte. A high-purity graphite rod was used as the working electrode, a platinum wire electrode was used as the counter electrode, and a calomel electrode was used as the reference electrode, and the working potential was 3V. After reacting for 2 hours, the solution was filtered through a 0.22 μm filter membrane to remove graphene sheets. Dialyze the above solution through a 3500Da dialysis bag for 24 hours to remove Na in the solution + and B 4 o 7 2- , to get B-GQDs. Gained B-GQDS is evenly distributed, and its particle size is between 3-8nm ( figure 1 ); Its elemental analysis found that it contains B element, the content is 3.2% ( figure 2 ).

[0038]...

Embodiment 3

[0045] Embodiment 3 establishes the working curve that detects HeLa cell concentration

[0046] Take 1.0 mL of reagent A and 20.0 μL of reagent B, mix well, add 10.0 μL of reagent C to make a mixed reagent, mix the mixed reagent with different concentrations of cervical cancer cell HeLa (10-10 6 cell / mL) at 37°C, 5% CO 2 Incubate for 2 h in the incubator; record the fluorescence signal F of B-GQDs by fluorescence spectrometer i ( Figure 4 ), Figure 4 The middle a-g respectively represent the cell concentration of 0, 10, 10 2 、10 3 、10 4 、10 5 、10 6 cell / mL. Calculate the difference ΔF of the response signal (ΔF=F i -F 0 ); The concentration of described ΔF and HeLa cells is drawn into ΔF-c working curve ( Figure 5 ); thereby established the HeLa cell concentration detection working curve based on the fluorescence response of B-GQDs; its linear range is 10-10 6 cell / mL.

Embodiment 4

[0047] Detection of HeLa cells in the sample to be tested in embodiment 4

[0048] Get the HeLa cell to be tested, act on the reagent in the kit according to the same method as in Example 3 and detect its fluorescence signal, the experimental results show that when the HeLa cell exists, the fluorescence signal of the quantum dots obviously recovers; according to the fluorescence signal of the HeLa cell The degree of change, from the standard curve obtained in Example 3, can further obtain the concentration of HeLa cells to be tested. Human embryonic kidney epithelial HEK 293T cells were used as the detection object. When it interacted with the kit, there was no significant change in the fluorescence signal of the quantum dots. It shows that the kit can be used for the specific detection of cells (such as HeLa cells) positively expressing alkaline phosphatase.

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Abstract

The invention discloses a cervical cancer cell detection kit based on a boron-doped graphene quantum dot and belongs to the field of biological detection. The cervical cancer cell detection kit comprises reagents A, B and C. The reagent A is a boron-doped graphene quantum dot solution, the reagent B is a cerium nitrate solution, and the reagent C is an adenosine triphosphate solution. Alkaline phosphatase is selected for the kit to serve as a detected targeted goal, the boron-doped graphene quantum dot is adopted as a fluorescent probe, alkaline phosphatase on the surfaces of cervical cancer cells is detected quickly and efficiently through a quantum dot fluorescence analysis method, and the kit can be used in the medical field to detect the cervical cancer cells and concentration thereof. The kit has the advantages of being easy to operate, high in detection result accuracy and sensitivity and low in cost and the like.

Description

technical field [0001] The invention relates to a detection kit, in particular to a detection kit for cervical cancer cells, belonging to the technical field of biological detection. Background technique [0002] Cervical cancer is a malignant tumor that seriously threatens women's health, and its incidence is increasing year by year. According to statistics, there are about 500,000 new cases of cervical cancer in the world every year, and about 260,000 women die from the disease. 85% of new cases occur in developing countries, among which the annual new cases of cervical cancer in my country account for about 1 / 3 of the total number of cases in the world. Therefore, the development of efficient, convenient, and highly sensitive cervical cancer detection methods is of great significance for the early diagnosis of cervical cancer. [0003] Traditional cervical cancer diagnostic methods include cervical iodine test, colposcopy, cervical photography, cervical exfoliated cytol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/42C12Q1/04
CPCG01N33/5044G01N2333/916
Inventor 吴萍陈力蔡称心
Owner NANJING NORMAL UNIVERSITY
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