APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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2024
·Vol. 108
·期 1
·1-12
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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
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LUMINESCENCE
·
2013
·Vol. 29
·期 4
·338-343
·
IF 1.273
Yuan Zhou,Tao Zhou,Rui Zhou,Yonggang Hu
A rapid, simple, facile, sensitive and enzyme-amplified chemiluminescence immunoassay (CLIA) method to detect antibodies against porcine parvovirus has been developed. Horseradish peroxidase (HRP) and the detection antibody were simultaneously co-immobilized on the surface of gold nanoparticles using the electrostatic method to form gold nanoparticle-based nanoprobes. This nanoprobe was employed in a sandwich-type CLIA, which enables CL signal readout from enzymatic catalysis and results in signal amplification. The presence of porcine parvovirus infection was determined in porcine parvovirus antibodies by measuring the CL intensity caused by the reaction of HRP–luminol with H 2 O 2 . Under optimal conditions, the obtained calibration plot for the standard positive serum was approximately linear within the dilution range of 1:80 to 1:5120. The limit of detection for the assay was 1:10,240 ( S / N = 3), which is much lower than that typically achieved with an enzyme-linked immunosorbent assay (1:160; S / N = 3). A series of repeatability measurements using 1:320-fold diluted standard positive serum gave reproducible results with a relative standard deviation of 4.9% ( n = 11). The ability of the immunosensor to analyze clinical samples was tested on porcine sera. The immunosensor had an efficiency of 90%, a sensitivity of 93.3%, and a specificity of 87.5% relative to the enzyme-linked immunosorbent assay results. Copyright © 2013 John Wiley & Sons, Ltd.
研究领域:
兽医学
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纳米技术
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免疫学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CLINICA CHIMICA ACTA
·
2023
·
IF 5
Ying Xu,Chenguang Hu,Jiayu liu,Haoran Guo,Jingkai Gao,Chi Wang,Chengbin Wang
Background and Aims Synovial fluid lactoferrin (LTF) and S100 calcium-binding protein A8 (S100A8) have been considered as potential biomarkers for the diagnosis of periprosthetic joint infection (PJI) through our previous research. However, the detection methods of these two proteins are still immature, so a rapid, accurate and cost-effective testing method is warranted. Materials and Methods We developed chemiluminescent immunoassays (CLIA) for the automated detection of synovial fluid LTF and S100A8 and assessed the analytical performance for these two methods. In addition, we recruited 86 patients who were suspected of PJI after total joint replacement (TJA) and examined their synovial fluid using CLIA to explore the clinical application value of these methods and the diagnostic efficiency of synovial fluid LTF and S100A8 for PJI. Results Our established CLIA methods have a wide linear range of 20–10,000 ng/mL for LTF detection system and 5–5000 ng/mL for S100A8 detection system. Performance parameters such as precision, specificity, and recovery rate can meet the industry standards. Then, the established methods were used to detect LTF and S100A8 in synovial fluid samples, which showed excellent diagnostic efficiency for PJI, and the areas under ROC curve (AUC) were 0.954 (95 % CI : 0.909–0.999) and 0.958 (95 % CI : 0.918–0.997), respectively. Conclusion Our established CLIA methods have the advantages of automation, high throughput, low price, and is expected to be widely popularized in clinical applications. Synovial fluid LTF and S100A8 detected through CLIA had efficient diagnostic potentiality for predicting and diagnosing PJI.
研究领域:
临床诊断
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骨科
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免疫学
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生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2019
·
IF 3.206
Zhi Zhang,Yueping Guan,Guoli Xu,Chen Guo
In this submission, a novel washing-free immunosensor was designed for chemiluminescence immunoassay (CLIA) of tumor marker using human Alpha-fetoprotein (AFP) as a representative. Reactive groups were introduced on the surface of the solvothermally prepared Fe 3 O 4 magnetic submicron particles (MSPs) and the dual-functional polyethylene glycol (PEG) linkers were modified to reduce the non-specific adsorption capacity of the particle surface and further to immobilize N -(4-aminohexyl)- N -ethylisoluminol (AHEI) and anti-AFP antibody on the surface of the particles. After incubation with AFP and horseradish peroxidase (HRP) labeled anti-AFP antibody respectively, the formed sandwich-type immunocomplexes on the particle surface could lead to the close proximity between AHEI and HRP, thus eventually leads to an increase in the detected chemiluminescence (CL) signal. As a bound HRP labeled anti-AFP antibody produces much stronger CL signal than an unbound one, this approach eliminates the time- and labor-consuming washing steps usually required of such assays. Under the optimal conditions, this present method shows a wide quantitative range from 1 to 250 ng mL −1 with a detection limit of 1.751 ng mL −1 . The detection limit was sufficient for measuring physiological (0–20 ng mL −1 ) and clinical (>20 ng mL −1 ) AFP levels. This method can also be applied in serum samples, and also avoid the problem of particle aggregation caused by multiple separation and washing steps.
研究领域:
生物医学工程
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癌症诊断
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分子生物学
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分析化学
✓ 已验证
产品关联
DOI
高影响因子
EUROPEAN FOOD RESEARCH AND TECHNOLOGY
·
2020
·Vol. 246
·期 9
·1857-1865
·
IF 2.366
Yang Fanfan,Xu Long,Liu Xiang,Chen Chen,Dias Alberto C. P.,Zhang Xiaoying
Abstract Bisphenol S (BPS) is a major alternative to Bisphenol A (BPA) newly applied in food packing material and disrupts reproductive and developmental endpoints through endocrine pathways. This study aimed to develop a rapid, sensitive, and accurate chemiluminescence immunoassay (CLIA) for the detection of BPS in canned beverage. Anti-BPS monoclonal antibody (mAb) was generated by immunizing Balb/c mice with immunogen (BPS–BSA conjugation). CLIA assay conditions, including 3-[9-(((3-( N -succinimidyloxycarboxypropyl) [4-methxylphenyl] sulfonyl) amine) carboxyl]-10-acridiniumyl)-1-propanesulfonate inner salt (NSP-SA-NHS) labeling ratio (1:40), antigen concentration (1 μg mL −1 ) and antibody titer (1:8000; 125 ng mL −1 in PBS (0.01 M, pH 7.4)) were determined, respectively. The half inhibition concentration (IC 50 ), detection limit (IC 10 ), and working range of the CLIA in canned beverage were 0.04, 1.48, and 0.32–5 ng mL −1 , respectively. Cross-reactivity with other bisphenols (4,4′-thiodiphenol, Bisphenol F, 4,4′- dihydroxy benzophenone, Bisphenol A and Bisphenol B) was less than 0.56%, and the recovery rates of BPS in canned beverage ranged from 80 to 109.2% (intra-assay) and from 82 to 108.4% (inter-assay). The results of CLIA were compared with high-performance liquid chromatography–UV (HPLC–UV) analysis, showing a good correlation of 0.956. After proper sample pre-treatment, this method allows relatively large amounts of sample analysis in a limited time period (30 min). Thus, it was deemed suitable and efficient for routine screening of BPS in canned beverage. Graphic abstract
研究领域:
分析化学
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环境科学
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食品安全
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免疫学
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毒理学
✓ 已验证
DOI
高影响因子
NANO
·
2018
·
IF 1.1
Yongfeng Gao,Yuanhui Feng,Lu Zhou,Lucia Petti,Zhe Wang,Jun Zhou,Shusen Xie,Jian Chen,Yanping Qing
Ultrasensitive detection of alpha-fetoprotein (AFP) is critical for the early diagnosis of liver cancer. In this work, a novel surface-enhanced Raman scattering (SERS)-based immunoassay complex has been successfully developed for the detection of AFP by using the Au-Ag alloy nanoparticals and the Ag/AgBr hybrid nanostructure. As the typical bimetal or metal/semiconductor plasmonic materials, besides the strong SERS enhancement characteristics, the Au-Ag alloy nanoparticals exhibit excellent monodispersity and the Ag/AgBr hybrid nanostructure demonstrates good stability. The experimental results show that the SERS-based immunoassay of AFP presents a low limit of detection of 1.86 fg/mL and a broad dynamic range from 2 fg/mL to 0.8 μ g/mL. Furthermore, the clinical applicability of the proposed SERS-based immunoassay has been assessed by the detection of AFP in the human serum samples of cancer patient and healthy person. The test data are consistent well with that of chemiluminescence immunoassay (CLIA) in the relative errors of − 8.82–8.06% and show better detection sensitivity. It reveals that the proposed immunoassay protocol is significant for giving insight into the design of ultrasensitive biosensor and the point-of-care testing of cancers.
研究领域:
分析化学
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生物医学工程
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纳米技术
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癌症研究
·
临床诊断
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）