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Electrochemical aptamer sensor for detecting mycobacterium tuberculosis MPT64 and detection method thereof

A technology of Mycobacterium tuberculosis and MPT64, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of expensive, time-consuming, poor sensitivity and specificity of sputum culture, etc.

Active Publication Date: 2022-03-29
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the clinical detection methods of Mycobacterium tuberculosis have their own defects, and it is difficult to fully realize the simple, rapid and accurate batch detection of Mycobacterium tuberculosis
For example: sputum culture has poor sensitivity and specificity; interferon-γ release assay, T-SPOT. Mycobacterium genetic tests (such as Xpert MTB / RIF test and line probe LPA test) can achieve rapid and accurate detection of M. tuberculosis infection, but they are expensive and require specialized technicians and expensive equipment

Method used

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  • Electrochemical aptamer sensor for detecting mycobacterium tuberculosis MPT64 and detection method thereof
  • Electrochemical aptamer sensor for detecting mycobacterium tuberculosis MPT64 and detection method thereof
  • Electrochemical aptamer sensor for detecting mycobacterium tuberculosis MPT64 and detection method thereof

Examples

Experimental program
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Embodiment 1

[0044] The main chemical reagent used in the embodiment of the present invention is as follows:

[0045] Recombinant MPT64 protein was purchased from Detai Biotechnology Co., Ltd. (Nanjing, China). AQ-2-COOH and bovine serum albumin (BSA) were purchased from Bailingwei Technology Co., Ltd. (Beijing, China). Streptavidin, chloroauric acid (HAuCl 4 ), tris(2-carboxyethyl)phosphine hydrochloride (TCEP) were purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). CMK-3 was purchased from Frontier Nanomaterials Technology Co., Ltd. (Nanjing, China). Sodium borohydride (NaBH 4 ) were purchased from Chengdu Kelong Chemical Reagent Factory (Chengdu, China). N,N-Dimethylformamide (DMF) was purchased from McLean Biochemical Technology Co., Ltd. (Shanghai, China). All DNA sequences in the experiment were synthesized and purified by Shanghai Sangon Co., Ltd., and the specific sequences are shown in Table 1.

[0046] Table 1. Nucleotide synthesis sequence list invo...

Embodiment 2

[0064] Example 2 uses electrochemical aptasensor to detect MPT64 antigen.

[0065] Using the electrochemical aptasensor constructed in Example 1 to detect the MPT64 antigen, follow the steps below:

[0066] (1) Drawing of standard curve:

[0067]The electrode of the biosensor constructed in Example 1 was placed in 0.1M PBS solution (containing 10mM KCl and 2mM MgCl 2 ), and measure the current values ​​of different concentrations of MPT64. A standard curve was drawn according to the logarithmic value of different concentrations of MPT64 and the DPV response signal. The test results showed that the two had a good linear relationship in the concentration range of 100fg / mL-10ng / mL, the linear correlation coefficient was 0.9885, and the detection limit was 67.6fg / mL. See the results for details figure 2 .

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Abstract

The invention provides an electrochemical aptamer sensor for detecting a specific antigen MPT64 of mycobacterium tuberculosis. The invention discloses a sandwich type electrochemical aptamer sensor which takes a nano compound CMK-3 / AQ-2-COOH formed by ordered mesoporous carbon CMK-3 and anthraquinone-2-carboxylic acid (AQ-2-COOH) as a signal response and amplification material, and takes Ce-MOF / AuNPs formed by a sea urchin-shaped cerium-based metal organic framework (Ce-MOF) and gold nanoparticles (AuNPs) as a sensing interface. The kit is used for detecting the specific antigen MPT64 of the mycobacterium tuberculosis pathogen of tuberculosis. Results show that Ce-MOF / AuNPs is used as a sensing interface, CMK-3 / AQ-2-COOH is used as a redox probe, signals of the sensor are amplified in a coordinated mode, ultra-sensitive detection of the specific antigen MPT64 of the mycobacterium tuberculosis is achieved, and a new detection method is provided for diagnosis of tuberculosis.

Description

technical field [0001] The invention belongs to the technical field of electrochemical aptamer sensors and nanocomposite materials, and in particular relates to an electrochemical aptamer sensor for detecting Mycobacterium tuberculosis specific antigen MPT64 and a detection method thereof. Background technique [0002] Tuberculosis is a serious infectious disease caused by Mycobacterium tuberculosis, which is transmitted through respiratory aerosols. According to the World Health Organization (WHO) 2020 Global Tuberculosis Report, there were about 10 million new cases of tuberculosis (TB) worldwide in 2019, and about 1.4 million people died of TB (including 208,000 HIV-infected people). Early diagnosis of tuberculosis is particularly important for tuberculosis treatment and infection control. [0003] At present, the clinical detection methods of Mycobacterium tuberculosis have their own defects, and it is difficult to completely realize the simple, rapid and accurate batch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30G01N27/327G01N27/48
CPCG01N27/308G01N27/3278G01N27/48Y02A50/30
Inventor 郭述良彭登勇白丽娟
Owner CHONGQING MEDICAL UNIVERSITY
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