MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 9
·1-9
·
IF 5.705
Gou Dan,Xie Guoming,Li Yuxia,Zhang Xin,Chen Hui
The authors describe an electrochemical immunoassay for ultrasensitive determination of the Mycobacterium tuberculosis (MTb) secretory protein MPT64 which is an antigen for early diagnosis of infection with MTb . Protein G was used to immobilize antibodies against MPT64 on a gold electrode. Graphene oxide with its large surface area was used as a carrier to anchor magnetite (Fe 3 O 4 ) and platinum (Pt) nanoparticles. The nanocomposite of type GO@Fe 3 O 4 @Pt was used as a signal reporter with excellent catalytic activity and recyclability. The nanocomposite exhibits peroxidase-like activity for hydrogen peroxide, best at −0.25 V (vs. Ag/AgCl). It works over a wide range of pH values (2–10) and temperatures (25–65 °C). Further signal amplification strategy was accomplished by rolling circle amplification. This voltammetric assay has a linear response to the logarithm of MPT64 concentration in the range from 5.0 fg·mL −1 to 1.0 ng·mL −1 and a detection limit of 0.34 fg·mL −1 (at a signal to noise ratio of 3, for n = 10). The assay can be completed within 4 h. It was successfully applied to the determination of MPT64 in spiked serum samples. Conceivably, the assay has a large potential in providing laboratory evidence for rapid diagnosis of MTb infection.
An electrochemical biosensor was developed for rapid detection of Mycobacterium tuberculosis secretory protein MPT64. The detection signal was synergistically amplified by GO@Fe 3 O 4 @Pt and rolling circle amplification. Phi29: phi29 DNA polymerase; BSA: bovine serum albumin; GO: graphene oxide.
研究领域:
医学诊断
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生物传感
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2022
·
IF 5.76
Yishi Li,Dengyong Peng,Shuliang Guo,Bijun Yang,Jing Zhou,Jiaxu Zhou,Qifan Zhang,Lijuan Bai
Rapid and sensitive tuberculosis (TB) diagnoses remain big challenges to current detection tools. In this work, a sensitive electrochemical aptasensor was constructed for the determination of Mycobacterium tuberculosis antigen MPT64 using a new redox nanoprobe. We found that anthraquinone derivative, anthraquinone-2-carboxylic acid (AQCA), a redox mediator, could be confined in ordered mesoporous carbon material of CMK-3. Due to the large loading amount of AQCA , as well as the confined space and electron transfer promotion effect of CMK-3, the obtained AQCA/CMK-3 nanohybrid with mass ratio of 2:1 showed excellent electroactivity and was employed as a new redox nanoprobe for signal amplification for the first time. Additionally, urchin-like Ce-MOFs were used to load a large amount of deposited gold nanocrystals (dep-Au), leading to dense immobilization of capture probe. The proposed electrochemical aptasensor for MPT64 detection showed a good linear relationship in the range from 100 fg/mL to 10 ng/mL with a low detection limit of 67.6 fg/mL. Besides, the aptasensor was utilized to detect MTP64 in human serum samples for TB diagnosis and presented satisfactory results.
研究领域:
生物医学工程
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电化学
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纳米技术
·
诊断学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOMATERIALS
·
2019
·
IF 10.273
Yuhan Chen,Xinzhu Liu,Shuliang Guo,Jun Cao,Jing Zhou,Jianli Zuo,Lijuan Bai
The present work described a novel sandwich-type electrochemical aptasensor for rapid and sensitive determination of Mycobacterium tuberculosis MPT64 antigen. Herein, a novel carbon nanocomposite composed of fullerene nanoparticles , nitrogen-doped carbon nanotubes and graphene oxide (C 60 NPs-N-CNTs/GO) was facilely synthesized for the first time, which not only possessed a large specific surface area and excellent conductivity , but also exhibited outstanding inherent electroactive property, and therefore served as nanocarrier and redox nanoprobe simultaneously. Gold nanoparticles (AuNPs) was then uniformly anchored onto the surface of such nanocomposite via Au–N bonds to bind with MPT64 antigen aptamer Ⅱ (MAA Ⅱ), forming the tracer label to realize generation and amplification of electrochemical signal. Additionally, conductive polyethyleneimine (PEI)-functionalized Fe-based metal-organic framework (P-MOF) was used as a sensing platform to absorb bimetallic core-shell Au–Pt nanoparticles ( [email protected] ), which could accelerate electron transfer and increase the immobilization of MPT64 antigen aptamer Ⅰ (MAA Ⅰ). After the typical sandwich-type protein-aptamer recognition, the inherent electroactivity of the tracer label was provoked by tetraoctylammonium bromide (TOAB), leading to a well-defined current response. Under the optimum condition, the proposed aptasensor showed a wide linear range for MPT64 detection from 1 fg/mL to 1 ng/mL with a limit of detection (LOD) as low as 0.33 fg/mL. More importantly, it was successfully used for MPT64 antigen detection in human serum, exhibiting a promising prospect for TB diagnosis in clinical practice.
研究领域:
分析化学
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纳米技术
·
生物医学工程
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电化学
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生物传感器
·
医疗诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 12
·1-7
·
IF 5.705
Li Ningning,Huang Xing,Sun Duanping,Yu Weiye,Tan Weiguo,Luo Zhaofan,Chen Zuanguang
An ultrasensitive aptasensor is described for the voltammetric determination of the Mycobacterium tuberculosis antigen MPT64 in human serum. Firstly, an amino-modified Zr(IV) based metal-organic framework (MOF; type UiO-66-NH 2 ; made up from Zr 6 O 32 units and 2-amino-terephthalate linkers) with a high specific surface was synthesized and used as the carrier of the gold nanoparticles and the aptamers. Then the signalling nanoprobe was fabricated after the horseradish peroxidase was cast on the nanomaterials. The two aptamers with synergistic effect on binding MPT64 were anchored on the gold electrode. Differential pulse voltammetry indicated that the peak current is highest if the ratio of the two aptamers is 1:1. The assay has a wide linear response range (0.02 to 1000 pg·mL −1 of MPT64) and a 10 fg·mL −1 detection limit at a working potential of around −96 mV (vs Ag/AgCl). The results show this biosensor to be a viable tool for detection of tuberculosis at an early stage.
Schematic presentation of the construction of the nanoprobe and biosensor. Firstly, the surface of UiO-66-NH 2 was anchored to gold nanoparticles (AuNPs). A dual-aptamer and HRP were added to form the signalling nanoprobe (Aptamer/HRP/AuNPs/UiO-66-NH 2 ). Then, the aptamers I and II were attached on the surface of gold electrode and 6-mercapto-1-hexanol was used to block the uncovered active site of the gold electrode. Finally, after incubation with MPT64, the signalling nanoprobe was dropped on the modified electrode and the DPV measurements was used for the analysis of Mycobacterium tuberculosis antigen MPT64. (PVP: poly(vinyl pyrrolidone); HRP: horseradish peroxidase; MCH: 6-Mercapto-1-hexanol; HQ: hydroquinone; BQ: benzoquinone).
研究领域:
生物传感器
·
电化学
·
纳米技术
·
结核病诊断
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）