MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 4
·1-10
·
IF 5.3
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
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生物传感技术
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材料科学
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2024
·Vol. 108
·期 1
·1-12
·
IF 3.9
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.
研究领域:
兽医学
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免疫学
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分子生物学
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诊断技术
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2022
·Vol. 106
·期 7
·2703-2714
·
IF 5.56
Geng Rui,Sun Yaning,Li Rui,Yang Jifei,Ma Hongfang,Qiao Zixuan,Lu Qingxia,Qiao Songlin,Zhang Gaiping
Abstract African swine fever virus (ASFV) causes a highly contagious and often lethal swine viral disease, and leads to tremendous economic losses to the swine industry. Unfortunately, there are no vaccines and effective antiviral agents available to prevent and control ASFV outbreaks. Therefore, it is necessary to develop simple and rapid strategies to monitor ASFV-infected pigs to restrain its spread. In the current study, ASFV capsid protein p72 was expressed along with its chaperone pB602L to form trimers in human embryonic kidney 293 (HEK293) cells. The p72 trimers were subsequently labeled with colloidal gold to develop a immunochromatographic strip. The strip showed high specificity to ASFV-positive serum and no cross-reactivity to other swine virus positive sera. Importantly, the strip showed a higher sensitivity of detecting ASFV antibodies in both positive standard serum and clinical serum samples than a commercial enzyme-linked immunosorbent assay (ELISA) kit. Taken together, these results demonstrate the strip as a reliable diagnostic tool against ASFV infection, which will be appropriate for application in prevention and control of ASFV.
Key points
• ASFV p72 trimers were successfully generated . • A colloidal gold strip was developed based on ASFV p72 trimers . • The strip is appropriate for detecting ASFV antibodies in the field .
研究领域:
兽医学
·
分子生物学
·
诊断技术
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2021
·Vol. 105
·期 11
·4751-4759
·
IF 4.813
Chen Dongjie,Wang Di,Wang Caixia,Wei Fang,Zhao Hongyuan,Lin Xiangmei,Wu Shaoqiang
Abstract Infection with African swine fever virus (ASFV) causes an acute and highly lethal hemorrhagic disease that has been responsible for huge economic losses in China. To exactly detect the antigen of ASFV, we established a rapid, no-wash, one-step sandwich-type immunoassay based on the amplified luminescent proximity homogenous assay-linked immunosorbent assay (AlphaLISA) using two monoclonal antibodies (mAbs) M-5 and M-6 against ASFV p72. ASFV p72 in samples was captured by biotinylated mAb M-5 connected to the donor bead surface via streptavidin and “sandwiched” by mAb M-6 which was coated onto the acceptor bead. Efficacy and sensitivity trials revealed that the AlphaLISA could detect ≥0.78 ng/ml of purified p72 and with a linear range of 0.78–100 ng/ml. The AlphaLISA was specific for ASFV and did not cross-react with other common pathogenic porcine viruses. Compared with RealPCR ASFV DNA test and ASFV antigen detection kit, the sensitivity of the AlphaLISA evaluated in 60 porcine serum samples was 93% and 100%, respectively. The specificity was 100% and 91.7%, respectively. This study presents a good laboratory diagnostic tool for sensitive and efficient detection of ASFV in porcine serum. Key points • MAbs M-5 and M-6 recognized various epitopes of ASFV p72. • The established ASFV p72 AlphaLISA showed well specificity, high sensitivity, and satisfied correlation coefficient.
研究领域:
兽医学
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分子诊断学
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免疫学
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 2
·68
·
IF 5.3
Guo Xing,Liu Yuanhang,Zhao Dong,Duan Qianqian,Sang Shengbo
African swine fever (ASF) is an acute hemorrhagic disease in pigs caused by the African swine fever virus (ASFV), which has a high mortality rate and brought great damage to global pig farming industry. At present, there is no effective treatment or vaccine to combat ASFV infection, so early detection of ASFV has become particularly important. Therefore, the PDMS/chitosan/MPMs composite film was proposed to detect ASFV P72. PDMS mixed with chitosan particles were used to provide biological modification sites for the ASFV P72 antibody, which simplified the functional modification process. The specific binding of antigen and antibody leads to the mechanical deformation of the composite film, which was observed through optical method to change the microsphere spacing. The polystyrene microsphere (PMs) spacing changes were acquired by simply measuring the diameters of the Debye ring diffracted by the PDMS/chitosan/PMs with a simple laser pointer. The conductive layer formed by AgNWs realize the transformation of biological signals to electrical signals. By conjugating PMs with Fe 3 O 4 nanoparticles (MPMs) and applying a magnetic field, a magnetic-stress-electric coupling system has been constructed, which amplifies the deformation of the composite film and enhances the sensitivity of the flexible film. The experimental results showed that the flexible film has wide linear range of 10 ng/mL–100 μg/mL, and the optimal detection limit (LOD) is 0.059 ng/mL, which was about one order of magnitude lower than that without magnetic field. In addition, the flexible film showed a good stability, specificity, and selectivity. Graphical
研究领域:
生物传感器
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病毒学
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纳米复合材料
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光学检测
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柔性电子学
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兽医诊断
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产品关联
DOI
PubMed
高影响因子
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