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MICROCHIMICA ACTA · 2025 ·Vol. 192 ·期 4 ·1-10 · IF 5.3

A flexible magnetic DNA biosensor composed of AgNWs/hydrogel/PS/Fe3O4 for the detection of ASFV P72 protein gene fragment

Xue Mengjie,Sang Shengbo,Zhao Dong,Duan Qianqian,Guo Xing

African swine fever virus (ASFV) is a highly infectious and pathogenic DNA virus with a mortality rate of nearly 100%. Here, we developed a flexible biosensor based on a AgNWs/hydrogel/PS(polystyrene)/Fe 3 O 4 composite film for the detection of ASFV P72 protein gene fragments. A large number of carboxyl groups in the hydrogel provide modification sites for capture probes (cp) simplifying the preparation process of the biosensor. Combined with the excellent magnetic properties of Fe 3 O 4 , the excellent electrical conductivity of silver nanowires (AgNWs) and the unique optical properties of two-dimensional photonic crystals (2DPC), the flexible biosensor can convert biological signals into optical signals and electrical signals, and the micro-deformation of the film can be verified by simple optical methods. In order to improve the sensitivity, we introduce Fe 3 O 4 and apply an external magnetic field to amplify the thin film micro-deformation. The concentration of P72 protein gene fragment and the relative change rate of resistance showed a good linear relationship, and the linear equation is y  = − 0.00126x − 0.31729; the detection limit (LOD) is as low as 0.208 μM. The composite film was used to detect real serum samples, and the recovery of the composite film fluctuated in the range 91.89 to 103.19%, indicating that the composite film has practical application potential in clinical detection of ASFV. In addition, the biosensor also shows good biocompatibility, stability, and specificity. Graphical

研究领域: 病毒学 · 生物传感技术 · 材料科学
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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY · 2024 ·Vol. 108 ·期 1 ·1-12 · IF 3.9

Development of plate-type and tubular chemiluminescence immunoassay against African swine fever virus p72

Miao Chun,Shao Junjun,Yang Sicheng,Wen Shenghui,Ma Yunyun,Gao Shandian,Chang Huiyun,Liu Wei

Abstract African swine fever (ASF) is a highly contagious and fatal viral disease that has caused huge economic losses to the pig and related industries worldwide. At present, rapid, accurate, and sensitive laboratory detection technologies are important means of preventing and controlling ASF. However, because attenuated strains of African swine fever virus (ASFV) are constantly emerging, an ASFV antibody could be used more effectively to investigate the virus and control the disease on pig farms. The isolation of ASFV-specific antibodies is also essential for the diagnosis of ASF. Therefore, in this study, we developed two chemiluminescence immunoassays (CLIAs) to detect antibodies directed against ASFV p72: a traditional plate-type blocking CLIA (p72-CLIA) and an automatic tubular competitive CLIA based on magnetic particles (p72-MPCLIA). We compared the diagnostic performance of these two methods to provide a feasible new method for the effective prevention and control of ASF and the purification of ASFV. The cut-off value, diagnostic sensitivity (Dsn), and diagnostic specificity (Dsp) of p72-CLIA were 40%, 100%, and 99.6%, respectively, in known background serum, whereas those of p72-MPCLIA were 36%, 100%, and 99.6%, respectively. Thus, both methods show good Dsn, Dsp, and repeatability. However, when analytical sensitivity was evaluated, p72-MPCLIA was more sensitive than p72-CLIA or a commercial enzyme-linked immunosorbent assay. More importantly, p72-MPCLIA reduced the detection time to 15 min and allowed fully automated detection. In summary, p72-MPCLIA showed superior diagnostic performance and offered a new tool for detecting ASFV infections in the future. Key points • Two chemiluminescence immunoassay (plate-type CLIA and tubular CLIA) methods based on p72 monoclonal antibody (mAb) were developed to detect ASFV antibody. • Both methods show good diagnostic performance (Dsn (100%), Dsp (99.6%), and good repeatability), and p72-MPCLIA detects antibodies against ASFV p72 with high efficiency in just 15 min.

CAS / MDL: 4432-31-9
研究领域: 兽医学 · 免疫学 · 分子生物学 · 诊断技术
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