Analytical Methods
·
2016
·Vol. 8
·期 7
·1577-1585
·
IF 1.915
Tao Liao,Fang Yuan,Haoyang Yu,Zigang Li
Sepsis is a potentially fatal systemic body infection with a significant mortality rate worldwide. Procalcitonin (PCT) is a sensitive and specific biomarker for sepsis diagnosis, however, the traditional immune-gold labeling detection of PCT is semi-quantitative with relatively low sensitivity (>0.1 ng mL−1). Recently proposed techniques (such as electrochemical immunosensor, microarray, and total internal reflection based methods) require either a complex sensor or sophisticated equipment. Herein, we report a magnetic bead and enzyme-antibody labeled gold nanoparticle based ELISA strategy for sensitive PCT detection. This developed assay is sensitive enough to detect PCT at 20 pg mL−1, efficient for the direct human serum test. Notably, this assay specifically distinguishes PCT from other sepsis markers and the whole assay takes only 1.5 hours to finish.
研究领域:
医学诊断
·
免疫学
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2018
·
IF 5.123
Weiguo Wang,Junmin Li,Changzhi Dong,Yafang Li,Qiuye Kou,Jinwu Yan,Lei Zhang
Amplifying the signal of ELISA is important in the early disease diagnosis. Herein we report an ultrasensitive ELISA applying for the detection of hCG based on the assemble of AuNPs induced by functional PAMAM . The AuNP-PAMAM probe shows a competitive advantage sensitivity of 0.03 IU L −1 compared to traditional ELISA and mAb 1 -AuNP-HRP probe. The line range is ranged from 0.1 to 6.4 IU L −1 . Moreover, the precision and reproducibility and specificity of AuNP-PAMAM probe are also eligible for the detection of hCG. The assembled AuNPs was firstly used in the signal enhancement in immunoassay .
研究领域:
生物医学
·
纳米技术
·
诊断学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Yafei Wang,Haitao Wang,Yan Zhang,Wei Li,Sijia Gao,Jiyan Qiao,Lei Zou,Zhuo Zhao,Ying Guan,Yongjun Zhang
Background Ultra-sensitive detection of biomarkers is critical for disease diagnosis and other applications. Immunoassays, particularly enzyme-linked immunosorbent assay (ELISA), are commonly used for protein biomarker detection but often suffer from insufficient sensitivity at low concentrations. Enhancing the detection capacity of ELISA for early diagnosis is therefore essential. Results Herein, a simple and cost-effective strategy was developed to enhance ELISA performance by pre-enriching target proteins using molecularly imprinted polymers (MIPs). MIPs, synthesized with high absorption capacity and specificity, were prepared using TrF (Transferrin) as the template and polypeptide as crosslinker. Target protein was pre-enriched through large-volume adsorption followed by small-volume elution. This pre-enrichment process resulted in an 8.25-fold increase in protein concentration, significantly improving ELISA sensitivity and reducing the limit of detection (LOD) by nearly an order of magnitude. The enhanced detection performance was validated in terms of accuracy, precision, and linearity. Significance This method, which combines MIP-based protein enrichment with ELISA detection, offers a promising and cost-effective solution for enhancing ELISA performance. It holds great potential as a widely applicable strategy for the sensitive detection of biomarkers in disease diagnostics.
研究领域:
生物化学
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yarong Ren,Juan Wei,Ye Wang,Peng Wang,Yanwei Ji,Baoyuan Liu,Jianlong Wang,Gualberto González-Sapienza,Yanru Wang
Salmonella are major pathogens that cause foodborne diseases. In this work, a broad-spectrum Salmonella nanobody-01 (Nb-01) was isolated and applied in the development of a streptavidin-bridged sandwich ELISA (SAB-ELISA) for simultaneously identifying five Salmonella serovars , including Salmonella Enteritidis ( S. Enteritidis), Salmonella Typhimurium ( S. Typhimurium), Salmonella London (S. London), Salmonella Paratyphi B (S. Paratyphi B) and Salmonella Hadar (S. Hadar). In this work, streptavidin (SA) was utilized as a scaffold to directionally immobilize biotinylated nanobody (BiNb) and Salmonella was detected by phage-displayed nanobodies. The SAB-ELISA can be accomplished within 180 min with a limit of detection (LOD) of 6.31 × 10 3 colony forming units (CFU) mL −1 , 9.15 × 10 3 CFU mL −1 , 4.23 × 10 3 CFU mL −1 , 7.31 × 10 3 CFU mL −1 and 7.25 × 10 3 CFU mL −1 towards S. Typhimurium, S. Enteritidis, S. London, S. Paratyphi B and S. Hadar, respectively. In comparison of sandwich ELISA by passive immobilization of Nb-01 on polystyrene plate, the sensitivity was increased by around 6-fold, which confirmed the enhanced immobilization efficacy of SAB-ELISA. Furthermore, the feasibility of the assay for S. Typhimurium determination in actual samples was evaluated, showing excellent recovery, inter-day and intra-day precision.
研究领域:
微生物学
·
免疫学
·
食品科学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2019
·
IF 2.398
Lu Zeng,Shanshan Song,Qiankun Zheng,Pengjie Luo,Xiaoling Wu,Hua Kuang
Shrimp tropomyosin is one of the most important causes of shellfish allergy. The objective of this study was to develop a highly sensitive and specific immunoassay for the detection of shrimp tropomyosin in food. Ten monoclonal antibodies against shrimp tropomyosin were obtained by fusion and cell screening. We found two optimum monoclonal antibodies (mAbs); mAb 2 was used as the capture antibody, and HRP-labelled mAb 3 was used as the detection antibody. Using this pair of mAbs, a sandwich enzyme-linked immunosorbent assay (ELISA) was developed with the limit of detection (LOD) of 0.45 ng/mL. Cross-reactivity with other food allergens using this method was found to be negligible. An immunochromatographic assay strip was also developed for the rapid detection of shrimp tropomyosin with the LOD of 0.5 ng/mL. The results demonstrated that our developmental methods represent a useful tool for the detection of shrimp tropomyosin in food.
研究领域:
免疫学
·
食品安全
·
生物技术
✓ 已验证
产品关联
DOI
高影响因子
TALANTA
·
2013
·
IF 3.498
Si Li,Chen-Gang Guo,Lu Chen,Xiao-Yang Yin,Yi-Xin Wu,Liu-Yin Fan,Hui-Zhi Fan,Cheng-Xi Cao
Hemoglobin A 1c (HbA 1c ) has been proven to be a key biomarker for diabetes screening, and glutathiolation of HbA 1c (viz., GSS-HbA 1c ) has been identified. However, the impact of GSS-HbA 1c on the measurement of HbA 1c for diabetes screening has not been quantitatively assessed yet. To address the issue, the micropreparative capillary isoelectric focusing (cIEF) developed in our previous work was used for the high resolution separation and purification of hemoglobin (Hb) species. The main fractions of HbA 0 , HbA 3 and HbA 1c extracted from the developed cIEF were identified by validated Mono S method. The proposed GSS-HbA 1c fractions in the cIEF were pooled and identified by electrospray ionization mass spectrometry (ESI-MS). The HbA 1c enzyme-linked immunosorbent assay (ELISA) kit was employed for further quantitative analysis of GSS-HbA 1c . A total of 34 blood samples with HbA 1c levels from 4.2% to 13.4% were assessed via the above comprehensive strategy of IEF-HPLC-MS-ELISA. It was demonstrated that the HbA 1c levels detected by cation exchange LC were considerably influenced by the glutathiolation of Hb and the range of detected GSS-HbA 1c values was between 0.23% and 0.74%. The results and developed cIEF methods have considerable significances for investigation of diabetes and clinical diagnosis.
研究领域:
医学诊断
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2018
·
IF 4.946
Chun Wang,Keyu Xing,Ganggang Zhang,Meifang Yuan,Shaolan Xu,Daofeng Liu,Wenyao Chen,Juan Peng,Song Hu,Wei-Hua Lai
Escherichia coli O157:H7 ( E. coli O157:H7) is a potential threat to human health; thus, a rapid and sensitive method for detecting it is necessary. We designed a single-stranded DNA that contained an appended block and anchoring block. The appended block acted as a scaffold to prepare fluorescent Ag nanoclusters (AgNCs). The anchoring block contained Poly A, which bound with the surface of gold nanoparticles to quench the fluorescence of AgNCs. An interesting ELISA approach for detecting E. coli O157:H7 was established via fluorescent quenching of DNA-stabilized AgNCs by using a sandwich complex. The changes in fluorescence intensity of AgNCs were used to quantitatively detect E. coli O157:H7. The sensitivity for detecting E. coli O157:H7 reached 1.905 × 10 3 CFU/mL with a good linear range. Compared with conventional ELISA, the sensitivity of this technique increased by 30-fold. Moreover, this method demonstrated specificity and reproducibility and could be used in food samples.
研究领域:
生物技术
·
分析化学
·
微生物学
·
食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Chaochao Chen,Liang Luo,Jianzhong Shen,Zhanhui Wang,Yantong Pan
The enzyme-linked immunosorbent assay (ELISA) stands as one of the most frequently employed rapid detection techniques for both chemical and biological contaminants. Horseradish peroxidase (HRP) and hydrogen peroxide (H2O2), serving as signal generators, play indispensable roles throughout the entire ELISA process. However, HRP and H2O2 exhibit high sensitivity to elevated temperatures, hindering the broader utilization and transportation of ELISA. Hereby, being inspired by the cascade catalysis of glucose by glucose oxidase (GOx) and HRP as well as the advantageous properties of metal–organic frameworks (MOFs), we developed a cascade catalysis-based competitive fluorescent ELISA (CCF-ELISA) for mycotoxin detection in maize using zearalenone (ZEN) as a model. The results revealed the remarkable protective effects of MOFs on enzymatic activities. Based on upon characteristics, the built CCF-ELISA enabled the detection of ZEN with a half-maximal inhibitory concentration of 0.54 ng/mL and allowed a detection range from 0.19 to 1.51 ng/mL. When applied to naturally contaminated maize samples, the outcomes obtained from the CCF-ELISA showed a high level of comparability with high-performance liquid chromatography–tandem mass spectrometry, demonstrating the promising potential of the developed CCF-ELISA in the rapid detection for food security.
研究领域:
食品安全检测
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2020
·
IF 6.306
Yi-Fang Li,Zheng-Zhong Lin,Cheng-Yi Hong,Zhi-Yong Huang
Due to complex matrixes and specific reagent deficiency, the rapid detection of histamine is still a challenge to date. Based on the high peroxidase-like activity of iron-cobalt co-doped carbon dots, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established for histamine detection using the mimic enzyme labeled with histamine antibody (His-Ab). Through the competitive binding of the labeled His-Ab to solid-phase and sample antigens, histamine content was detected with a linear range of 2.5–150 μg mL −1 . The detection limit based on 3σ/K was 0.50 mg kg −1 , which was much lower than those of commercial His-kit and HPLC methods. The ic-ELISA method was applied to histamine detection in fish samples with the recovery of (103.4 ± 0.5)%, which was in accord with those of commercial His-kit and HPLC methods. The results indicated that the established ic-ELISA method was suitable for rapid detection of histamine in fish samples with high accuracy, sensitivity and stability.
研究领域:
食品科学
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2016
·Vol. 8
·期 17
·3578-3586
·
IF 1.915
Haijing Du,Yanxin Chu,Hong Yang,Kang Zhao,Jianguo Li,Pei She,Xiang Zhang,Anping Deng
Brombuterol, a new β-adrenergic agonist which can enhance the lean meat-to-fat ratio, is forbidden as an additive in animal feeds for livestock production due to its adverse effects on consumers. In this study, a highly sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA) for the detection of brombuterol in tissue and feed samples was developed. The immunogen and coating antigen were prepared by directly linking brombuterol to carrier proteins using the diazobenzidine method. The antisera against brombuterol were obtained from immunized rabbits. The IC50 and limit of detection (LOD) values of the superior ELISA were 0.165 ng mL−1 and 0.007 ng mL−1, respectively. The cross-reactivity (CR) values of ELISA with clenbuterol, terbutaline and cimbuterol were less than 6.2%; there was no CR of ELISA with nine other compounds. To investigate the accuracy and precision of the assay, liver, meat and feed samples were spiked with brombuterol at different concentrations and analyzed by ELISA. Acceptable recovery rates of 91.9–115.4% and intra-assay coefficients of variation of 1.5–9.5% (n = 3) were achieved. The ELISA was also validated by HPLC. As revealed, both methods were highly correlated (R2 = 0.9927–0.9994, n = 6). The proposed ELISA was proven to be a feasible method for quantitative analysis of brombuterol in tissue and feed samples.
研究领域:
食品安全
·
免疫学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2016
·
IF 2.857
Feng Wang,Hong Wang,Yu-Dong Shen,Yong-Jun Li,Jie-Xian Dong,Zhen-Lin Xu,Jin-Yi Yang,Yuan-Ming Sun,Zhi-Li Xiao
A new multianalyte immunoassay was designed to screen furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone (AMOZ), malachite green (MG), and leucomalachite green (LMG) in aquatic products using a bispecific monoclonal antibody (BsMAb). Gradient drug mutagenesis methods were separately used to prepare an anti-3-nitrobenzaldehyde-derivatized AMOZ (3-NPAMOZ) hybridoma cell line that was hypoxanthine-guanine-phosphoribosyltransferase (HGRPT) deficient and an anti-LMG hybridoma cell line that was thymidine kinase (TK) deficient. BsMAb recognizing 3-NPAMOZ and LMG was generated using hybrid–hybridomas of HGRPT and TK deficient cell lines. For AMOZ and LMG, respectively, the BsMAb-based indirect competitive ELSIA (ic-ELISA) values of 1.7 ng/mL and 45.3 ng/mL and detection limits of 0.2 ng/mL and 4.8 ng/mL. To establish the ic-ELISA, 3-NPAMOZ derivatized from AMOZ with 3-nitrobenzaldehyde and LMG reduced from MG by potassium borohydride was recognized by BsMAb. Recoveries of AMOZ, MG, and LMG in aquatic products were satisfactory and correlated with HPLC analysis. Thus, the multianalyte ic-ELISA is suitable for rapid quantification of AMOZ, MG, and LMG in aquatic products.
研究领域:
分析化学
·
食品科学
·
免疫学
·
水产养殖
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2015
·Vol. 7
·期 15
·6190-6197
·
IF 1.821
Xiaoling Wu,Dezhao Kong,Shanshan Song,Liqiang Liu,Hua Kuang,Chuanlai Xu
A pair of sensitive monoclonal antibodies against both Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola were generated by immunizing mice and cell fusion techniques. The sandwich enzyme-linked immunosorbent assay method and immunochromatographic test strip were developed based on newly generated monoclonal antibodies for detection in rice samples. The lower limit of detection of the sandwich ELISA method for X. oryzae pv. oryzae was 1.0 × 104 cfu mL−1. The cut-off values for the immunochromatographic test strip for X. oryzae pv. oryzae and X. oryzae pv. oryzicola in rice samples were 3.3 × 105 cfu mL−1 and 1 × 106 cfu mL−1, respectively. The quantitative sandwich ELISA method and rapid immunochromatographic test strip assay could be useful for food safety inspections.
研究领域:
植物病理学
·
微生物学
·
食品安全
✓ 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）