tetano
Editor, Senior Moderator
Anal Chim Acta. 2017 Apr 22;963:112-118. doi: 10.1016/j.aca.2017.01.031. Epub 2017 Jan 31.
[h=1]Highly sensitive colorimetric immunosensor for influenza virus H5N1 based on enzyme-encapsulated liposome.[/h] Lin C[SUP]1[/SUP], Guo Y[SUP]1[/SUP], Zhao M[SUP]1[/SUP], Sun M[SUP]1[/SUP], Luo F[SUP]2[/SUP], Guo L[SUP]1[/SUP], Qiu B[SUP]1[/SUP], Lin Z[SUP]1[/SUP], Chen G[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Development of simple but sensitive biosensor for influenza detection is highly important in immediate and effective clinical treatment. In this study, a sensitive colorimetric immunosensor which combines the advantages of high selectivity of immunoassay and simplicity of colorimetric detection has been developed to detect influenza virus H5N1 based on enzyme-encapsulated liposome. Biotin-tagged liposome encapsulated with large amount of horseradish peroxidase (HRP) was firstly synthesized. In the presence of H5N1, H5N1 co-bound with the capture antibody and the biotinylated detection antibody to form sandwich immunocomplex. Subsequently, the HRP-encapsulated liposome was introduced to conjugate with the detection antibody through biotin-avidin-biotin linkage. Upon the addition of substrate (mixture of 3,3',5,5'-tetramethylbenzidine (TMB) and H[SUB]2[/SUB]O[SUB]2[/SUB]), the liposome was directly lysed to release large amount of HRP by TMB. The released HRP catalyzed the H[SUB]2[/SUB]O[SUB]2[/SUB]-mediated oxidation of TMB, resulting in color change of the system, which was observed by naked eyes or UV-vis spectra. The result showed that the absorption intensity enhanced with the increase of H5N1 concentration ranging from 0.1 to 4.0 ng/mL, and the detection limit was calculated to be 0.04 ng/mL. The sensitivity of the proposed biosensor is much higher than that of conventional enzyme-linked immunosorbent assay method. The proposed immunosensor is relatively simple, low-cost, sensitive, and selective without using any sophisticated instruments, therefore it may have a promising prospect for detecting targets in clinical medicine, food safety analysis, and environmental monitoring.
Copyright ? 2017 Elsevier B.V. All rights reserved.
[h=4]KEYWORDS:[/h] Colorimetric; H5N1; Immunoassay; Liposome; Signal amplification
PMID: 28335964 DOI: 10.1016/j.aca.2017.01.031
[h=1]Highly sensitive colorimetric immunosensor for influenza virus H5N1 based on enzyme-encapsulated liposome.[/h] Lin C[SUP]1[/SUP], Guo Y[SUP]1[/SUP], Zhao M[SUP]1[/SUP], Sun M[SUP]1[/SUP], Luo F[SUP]2[/SUP], Guo L[SUP]1[/SUP], Qiu B[SUP]1[/SUP], Lin Z[SUP]1[/SUP], Chen G[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Development of simple but sensitive biosensor for influenza detection is highly important in immediate and effective clinical treatment. In this study, a sensitive colorimetric immunosensor which combines the advantages of high selectivity of immunoassay and simplicity of colorimetric detection has been developed to detect influenza virus H5N1 based on enzyme-encapsulated liposome. Biotin-tagged liposome encapsulated with large amount of horseradish peroxidase (HRP) was firstly synthesized. In the presence of H5N1, H5N1 co-bound with the capture antibody and the biotinylated detection antibody to form sandwich immunocomplex. Subsequently, the HRP-encapsulated liposome was introduced to conjugate with the detection antibody through biotin-avidin-biotin linkage. Upon the addition of substrate (mixture of 3,3',5,5'-tetramethylbenzidine (TMB) and H[SUB]2[/SUB]O[SUB]2[/SUB]), the liposome was directly lysed to release large amount of HRP by TMB. The released HRP catalyzed the H[SUB]2[/SUB]O[SUB]2[/SUB]-mediated oxidation of TMB, resulting in color change of the system, which was observed by naked eyes or UV-vis spectra. The result showed that the absorption intensity enhanced with the increase of H5N1 concentration ranging from 0.1 to 4.0 ng/mL, and the detection limit was calculated to be 0.04 ng/mL. The sensitivity of the proposed biosensor is much higher than that of conventional enzyme-linked immunosorbent assay method. The proposed immunosensor is relatively simple, low-cost, sensitive, and selective without using any sophisticated instruments, therefore it may have a promising prospect for detecting targets in clinical medicine, food safety analysis, and environmental monitoring.
Copyright ? 2017 Elsevier B.V. All rights reserved.
[h=4]KEYWORDS:[/h] Colorimetric; H5N1; Immunoassay; Liposome; Signal amplification
PMID: 28335964 DOI: 10.1016/j.aca.2017.01.031