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Sandwich type electrochemical immunosensor for simultaneously detecting markers of cervical cancer

An immunosensor, electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of low analysis efficiency, unsatisfactory sensitivity and specificity of tumor markers, etc., to achieve improved sensitivity, immune analysis Rapid and reproducible results

Active Publication Date: 2013-12-25
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In view of the unsatisfactory sensitivity and specificity of existing tumor markers, in order to improve the clinical application value of tumor markers, it is very important to select multiple markers for simultaneous detection according to the characteristics of different tumors and different markers.
Due to the low analysis efficiency for the detection of a single tumor marker, in order to shorten the analysis time, reduce the analysis steps, improve the analysis efficiency, and reduce the test cost, people began to pay attention to the high-throughput electrochemical immunosensor that can detect multiple samples simultaneously. Research

Method used

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  • Sandwich type electrochemical immunosensor for simultaneously detecting markers of cervical cancer
  • Sandwich type electrochemical immunosensor for simultaneously detecting markers of cervical cancer
  • Sandwich type electrochemical immunosensor for simultaneously detecting markers of cervical cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of Sandwich Electrochemical Immunosensor

[0037] (1) Prepare the secondary antibody incubated with AuCMK-3 and electron mediator (Ab 2 ) solution

[0038]Synthesis of AuCMK-3: 100 mL of a solution containing 0.01 g of chloroauric acid was heated to boiling in a flask, then 10 mL of 1% sodium citrate was added dropwise and the temperature was maintained for 40 min, then cooled to room temperature. Mesoporous carbon CMK-3 was added to the solution and stirred for 4h. Finally, centrifuge at 9000 rpm for 20 min, centrifuge three times, and dry in vacuum to obtain AuCMK-3; figure 1 It is the SEM photo of AuCMK-3, from which it can be seen that Au nanoparticles are uniformly dispersed on the surface of mesoporous carbon CMK-3;

[0039] Secondary antibody incubated with AuCMK-3 and electron mediator (Ab 2 ) solution: Weigh two parts of Au CMK-3 (4 mg) prepared in the above step ① in parallel, add PBS with a pH of 7.4 to dissolve it by ultrasonic, t...

Embodiment 2

[0049] Example 2 Simultaneous detection of cervical cancer tumor markers: SCCA and CEA

[0050] The electrochemical immunosensor was prepared according to the steps described in Example 1. The electrochemical immunosensor was used to simultaneously detect the cervical cancer tumor markers shown in Table 1. The detection technical indicators are shown in Table 1.

[0051] Table 1 Detection technical indicators of cervical cancer tumor markers

[0052]

[0053] It can be seen from the detection technical index results in Table 1 that the electrochemical immunosensor prepared by the present invention is used for the simultaneous detection of cervical cancer tumor markers, and has a wide linear range, low detection limit and high method sensitivity.

Embodiment 3

[0054] Example 3 Simultaneous detection of cervical cancer tumor markers in human serum

[0055] The concentrations of prepared CEA antigens were 0.05, 1.0, 5.0, 10, 15, 20 ng / mL and the concentrations of SCCA antigens were 0.03, 1.0, 5.0, 10, 15, 20 ng / mL in turn, and the corresponding working curve, respectively as figure 2 A and figure 2 As shown in B, it is used for the simultaneous detection of CEA and SCCA in human serum. Fresh serum samples were taken, and the supernatant was taken after centrifugation, diluted with PBS buffer solution, and the contents of CEA and SCCA in human serum were determined. Then, a certain concentration of CEA and SCCA standard solution was added to human serum, and the recovery experiment was carried out. The average recovery rate of CEA and SCCA in the sample was calculated by measuring the ratio of the recovery value and the added amount. The test results are shown in Table 2.

[0056] Table 2 Detection results of cervical cancer tum...

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Abstract

The invention relates to an electrochemical immunosensor, and in particular relates to a sandwich type electrochemical immunosensor which is constructed by taking Au-hybridized mesoporous carbon CMK-3(Au@CMK-3) and two electronic mediators, namely neutral red and thionine, as second antibody markers, a preparation method of the sandwich type electrochemical immunosensor, and application of the electrochemical immunosensor, which is prepared by the method, to simultaneous detection on markers of a cervical cancer. The electrochemical immunosensor can be used for simultaneously detecting two samples and overcoming the limitation of a single index, plays an important role in further improvement of the early confirming rate of the cervical cancer, and can be popularized to the field of simultaneous detection on tumor markers of other cancers.

Description

technical field [0001] The present invention relates to an electrochemical immunosensor, more specifically, the present invention relates to a mesoporous carbon CMK-3 (AuCMK-3) based on Au hybridization and two electron mediators of neutral red and thionine respectively as A sandwich type electrochemical immunosensor constructed with a secondary antibody marker, a preparation method thereof, and an application of the electrochemical immunosensor prepared by the method in simultaneous detection of cervical cancer markers. Background technique [0002] Cervical cancer is a common malignant tumor of the female reproductive system. Although the incidence of cervical cancer has decreased significantly in recent decades, there are still 131,500 new cases of cervical cancer in my country every year, accounting for about 28.8% of new cases of cervical cancer in the world. youth trend. Early and accurate detection of cervical precancerous lesions or cervical cancer is the key to redu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N33/574
Inventor 吴丹魏琴张勇郭爱平郭占魁
Owner UNIV OF JINAN
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