JOURNAL OF DAIRY SCIENCE
·
2016
·
IF 2.408
Shan Shan,Daofeng Liu,Qi Guo,Songsong Wu,Rui Chen,Kai Luo,Liming Hu,Yonghua Xiong,Weihua Lai
In this study, cascade signal amplification in ELISA involving double-antibody sandwich ELISA and indirectly competitive ELISA was established to sensitively detect Escherichia coli O157:H7. In the double-antibody sandwich ELISA, a complex was formed comprising anti- E. coli O157:H7 polyclonal antibody, E. coli O157:H7, biotinylated anti- E. coli O157:H7 monoclonal antibody, streptavidin, and biotinylated β-lactamase. Penicillin solution was then added into the ELISA well and hydrolyzed by β-lactamase. Afterward, the penicillin solution was transferred to indirectly competitive ELISA. The concentration of penicillin can be sensitively detected in indirectly competitive ELISA. In the cascade signal amplification system, increasing the amount of added E. coli O157:H7 resulted in more β-lactamase and less penicillin. The detection sensitivity of E. coli O157:H7, which was 20 cfu/mL with the cascade signal amplification in ELISA, was 1,000-fold higher than that of traditional ELISA. Furthermore, the novel method can be used to detect E. coli O157:H7 in milk (2 cfu/g). Therefore, this new signaling strategy will facilitate analyses of highly sensitive foodborne pathogens.
研究领域:
微生物学
·
食品科学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
International Journal of Environmental Research and Public Health
·
2013
·Vol. 10
·期 7
·2688-2698
·
IF 1.998
Wenbin Wang,Liqiang Liu,Liguang Xu,Wei Ma,Hua Kuang,Chuanlai Xu
β-Lactamase residues in milk represent a public health risk. The cylinder plate detection method, which is based on bacterial growth, is laborious and time consuming. In this study, 15 monoclonal antibodies (mAbs) were selected against Temoneira (TEM) 1 β-lactamase. A sandwich enzyme-linked immunosorbent assay (ELISA) based on an optimum mAb pair was developed and validated for the detection of β-lactamase. The limit of detection and linear dynamic range of the method were 4.17 ng/mL and 5.5–100 ng/mL, respectively. β-Lactamase recovery in pure milk was 96.82–103.13%. The intra- and inter-assay coefficients of variation were 6.21–7.38% and 12.96–13.74%, respectively. Our developed sandwich ELISA can be used as a rapid detection method of β-lactamase in milk.
研究领域:
食品科学
·
免疫学
·
公共卫生
✓ 已验证
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Yang Liu,Meng Cai,Weiping Wu,Yuting Fang,Pei She,Shouming Xu,Jianguo Li,Kang Zhao,Jingjuan Xu,Ning Bao,Anping Deng
Herein, we reported a disposable electroanalytical device for competitive enzyme-linked immunosorbent assay (ELISA) of phenylethanolamine A (PA). Conductive carbon tape coated with gold cluster served as the working electrode , while the counter and reference electrodes were fabricated on the filter paper by the screen-printing technique. Separating fabrication of the working electrode from other electrodes could make it possible for full utilization of the working electrode modification in bulk and no contamination of the immune-reagents on the counter and reference electrodes. The gold cluster played an important role in both immobilizing antigen and accelerating electron transfer . The eight-channel devices were utilized to detect PA on the strategy of competitive ELISA. The detection range and the limit of detection (LOD) using differential pulse voltammetry for PA were 0.005–60 ng mL −1 and 2.6 pg mL −1 , respectively. And also, the linear range for PA performed by square wave voltammetry was 0.05–60 ng mL −1 with the LOD value as 0.028 ng mL −1 . The results clearly demonstrated that the proposed electroanalytical devices could be successfully applied in immunoassay and might be further developed for determination of different analytes based on ELISA format.
研究领域:
分析化学
·
免疫学
✓ 已验证
DOI
PubMed
高影响因子
ANALYST
·
2016
·Vol. 141
·期 10
·2970-2976
·
IF 4.033
Yanyan Li,Xiaoming Ma,Zhengming Xu,Meihua Liu,Zhenyu Lin,Bin Qiu,Longhua Guo,Guonan Chen
Seed-mediated synthesis of gold nanorods (AuNRs) has been widely used for diverse applications in the past decade. In this work, this synthetic process is demonstrated for multicolor biosensing for the first time. Our investigation reveals that ascorbic acid acts as a key factor to mediate the growth of AuNRs. This phenomenon is incorporated into the alkaline phosphatase (ALP)-enzyme-linked immunosorbent assay (ELISA) system based on the fact that ALP can catalyze the conversion of ascorbic acid–phosphate into ascorbic acid with high efficiency. This allows us to develop a multicolor ELISA approach for sensitive detection of disease biomarkers with the naked eye. We show the proof-of-concept multicolor ELISA for the detection of prostate-specific antigen (PSA) in human serum. The results show that different colors are presented in response to different concentrations of PSA, and a detection limit of 3 × 10−15 g mL−1 in human serum was achieved. The proposed multicolor ELISA could be a good supplement to conventional ELISA for POC diagnostics.
研究领域:
生物化学
·
医学诊断
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2016
·
IF 2.687
Lu Li,Zheng-zhong Lin,Ai-hong Peng,Hui-ping Zhong,Xiao-mei Chen,Zhi-yong Huang
A direct competitive enzyme-linked immunosorbent assay (ELISA) method was used for the detection of malachite green (MG) with a high sensitivity and selectivity using magnetic molecularly imprinted polymers (MMIPs) as a bionic antibody. MMIPs were prepared through emulsion polymerization using Fe 3 O 4 nanoparticles as magnetic nuclei, MG as a template, methacrylic acid (MAA) as a functional monomer, ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent and span-80/tween-80 as mixed emulsifiers. The MMIPs were characterized by scanning electron micrographs (SEM), thermal-gravimetric analyzer (TGA), Fourier transform infrared spectrometer (FT-IR) and vibrating sample magnetometer (VSM), respectively. A high magnetic saturation value of 54.1 emu g −1 was obtained, resulting in rapid magnetic separation of MMIPs with an external magnet. The IC 50 of the established ELISA method was 20.1 μg L −1 and the detection limit (based on IC 85 ) was 0.1 μg L −1 . The MMIPs exhibited high selective binding capacity for MG with cross-reactivities less than 3.9% for MG structural analogues. The MG spiking recoveries were 85.0%–106% with the relative standard deviations less than 4.7%. The results showed that the biomimetic ELISA method by using MMIPs as bionic antibody could be used to detect MG rapidly in fish samples with a high sensitivity and accuracy.
研究领域:
分析化学
·
环境科学
·
食品科学
✓ 已验证
DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2014
·
IF 0.984
Juan Peng,Xu Meng,Xiaofang Deng,Jianping Zhu,Hua Kuang,Chuanlai Xu
Hen egg is one of the most frequent causes of food allergy in infants and adults. Ovalbumin (OVA) has been identified as a major egg allergen. In order to detect OVA in foods, a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) based on two monoclonal antibodies (mAbs) was established. The 2 mAbs were selected out of 17 murine hybridomas secreting OVA-specific antibody. Using mAb17 as the capture antibody and mAb15 as the detection antibody, the detection limit of the ELISA method was 0.51 ng/mL, and the linear dynamic range was between 1.95 and 500 ng/mL. The recovery ranged from 85.6 to 115.2%, whereas the intra- and inter-assay coefficients of variation were less than 8.6 and 13.9%, respectively. Sample analysis verified that the produced anti-OVA mAb and the developed ELISA may provide a valuable tool for the sensitive determination of OVA in processed foods and for future studies on the mechanism of how OVA functions in anaphylaxis.
研究领域:
免疫学
·
食品科学
·
医学
✓ 已验证
产品关联
DOI
高影响因子
International Journal of Environmental Research and Public Health
·
2013
·Vol. 10
·期 7
·2897-2905
·
IF 1.998
Juan Peng,Shanshan Song,Liguang Xu,Wei Ma,Liqiang Liu,Hua Kuang,Chuanlai Xu
We have established a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) based on two monoclonal antibodies (mAb) to measure the content of the major peanut allergen Ara h 1 in foods. Two mAbs were selected out of 12 murine hybridoma cells secreting Ara h 1-specific antibody. Using mAb 6 as the capture antibody and HRP-labelled mAb 4 as the detection antibody, the limit of detection (LOD) the assay was 0.34 ng/mL. Cross-reaction analysis showed that this method was strongly specific and had no cross-reactions with Ara h 2, pea protein or soy protein. Sample analysis showed that this ELISA was a useful tool to monitor peanut allergens in food products by measuring Ara h 1 content.Keywords:peanut allergen;Ara h 1;monoclonal antibody;sandwich ELISA
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
·
2014
·Vol. 6
·期 24
·9616-9621
·
IF 1.938
Chifang Peng,Xiaohui Duan,Gabriel Wafala Khamba,Zhengjun Xie
Based on the H2O2-mediated generation of gold nanoparticles, a colorimetric catalase nanosensor was developed and utilized as a signal output in a catalase-labelled immunoassay. Methyltestosterone (MT), which is used illegally as a growth promoter during the cattle production cycle, was selected as a small molecule model in this new ELISA development. As a result, a “signal on” plasmonic ELISA for MT was established. This plasmonic ELISA can quantitatively detect MT by spectrophotometry with a 50 times higher sensitivity than the traditional ELISA. In addition, it allows the highly sensitive and easily qualitative detection of MT by the naked eye. This plasmonic direct competitive ELISA can be utilized for developing cost-effective and sensitive ELISA against various small molecules by the naked eye.
研究领域:
分析化学
·
纳米技术
·
生物传感器
·
环境科学
·
食品安全
✓ 已验证
产品关联
DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2012
·
IF 0.714
Xiaofang Deng,Liqiang Liu,Wenwei Ma,Chuanlai Xu,Libing Wang,Hua Kuang
In this study, two anti-peanut agglutinin (PNA) polyclonal antibodies were successfully prepared. Using mouse anti-PNA polyclonal antibody as the capture antibody and rabbit anti-PNA polyclonal antibody as the detection antibody, a novel sandwich enzyme-linked immunosorbent assay (ELISA) was developed to quantify PNA. The detection limit of the ELISA method was low (0.49 ng/mL), and the linear dynamic range was between 0.78 and 100 ng/mL. The recovery ranged from 93.86% to 139.3%, whereas the intra- and inter-assay coefficients of variation were less than 10.25% and 12.06%, respectively. Sample analysis verified this method as a reliable tool for the detection of PNA in processed foods.
研究领域:
免疫学
·
食品科学
✓ 已验证
产品关联
DOI
高影响因子
Toxins
·
2018
·Vol. 11
·期 1
·6
·
IF 3.273
Ming Li,Mingna Sun,Xia Hong,Jinsheng Duan,Daolin Du
A total of 328 agricultural product samples highly suspected to be contaminated, from flour companies, feed companies, and livestock farms throughout China, were surveyed for deoxynivalenol (DON) contamination using a self-assembly enzyme-linked immunosorbent assay (ELISA) kit. An ELISA kit for DON was developed with a 4.9 ng mL−1limit of detection (LOD) in working buffer and a 200 ng g−1LOD in authentic samples. The DON contamination detection rate was 88.7%, concentrations ranged from 200.9 to 6480.6 ng g−1, and the highest DON contamination was found in distillers’ dried grains with solubles with an average of 3204.5 ng g−1. Wheat bran and wheat were found to be the most commonly contaminated samples, and the corn meal samples had the lowest average DON level. This ELISA kit is a powerful alternative method for the rapid, sensitive, specific, accurate, and high-throughput determination of DON and can meet the maximum requirement levels. This survey suggests that DON contamination in the Chinese market is serious, and the contamination risk deserves attention. Essential preventive measures should be implemented to ensure food safety and human health.Keywords:deoxynivalenol;contamination;survey;ELISA kit;agricultural productKey Contribution:A total of 328 agricultural product samples highly suspected to be contaminated were surveyed for deoxynivalenol using a self-assembly ELISA kit.
研究领域:
食品安全
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food Analytical Methods
·
2015
·Vol. 8
·期 10
·2605-2611
·
IF 1.956
Peng Juan,Song Shanshan,Liu Liqiang,Kuang Hua,Xu Chuanlai
In order to develop highly sensitive immunoassays for detecting the major peanut allergen, Ara h 2, we prepared six monoclonal antibodies (mAb) with high affinity to Ara h 2. Using mAb 5 as a capture antibody and HRP-labeled mAb 1 as a detection antibody, a sandwich enzyme-linked immunoassay (ELISA) was established with a limit of detection (LOD) of 0.02 ng/mL. An immunochromatographic strip based on these two mAbs was also established with a LOD of 1 ng/mL. Both tests showed high specificity in cross-reactivity experiments. Sample analyses indicated that these two immunoassays were feasible, accurate, and highly sensitive for measuring allergen Ara h 2 of food.
研究领域:
食品科学
·
免疫学
·
生物化学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2016
·
IF 4.712
Lin Luo,Hong-Tao Lei,Jin-Yi Yang,Gong-Liang Liu,Yuan-Ming Sun,Wei-Dong Bai,Hong Wang,Yu-Dong Shen,Sui Chen,Zhen-Lin Xu
The widespread occurrence of ethyl carbamate (EC, 89.09 Da), a group 2A carcinogen, in fermented foods and alcoholic beverages has raised worldwide public health concern. Immunoassay for EC is unavailable due to the simple and small structure of EC. In this work, an initial attempt to produce antibody specific for EC, by using 4-((ethoxycarbonyl)amino)butanoic acid as hapten , was made but failed. However, since EC can easily react with 9-xanthydrol to form xanthyl ethyl carbamate (XEC), two haptens based on XEC structure were designed and synthesized. Polyclonal antibody against XEC, instead of EC was obtained and then used to develop a competitive indirect ELISA for EC via a pre-analysis derivatization . After optimization, the ciELISA was applied in analyzing Chinese rice wine with detection limit of 166 μg/L, and negligible cross-reactivity with EC analogs. Recoveries of EC in fortified samples were from 84.4% to 100.9%, with coefficients of variation below 10%. Results for analysis of real samples by the ci-ELISA correlated well with that by reference method GC-MS, suggesting the good accuracy and reproducibility of the proposed method. This is the first report of an immunoassay capable of detecting EC, which is suitable for monitoring EC in a large amount of samples.
研究领域:
食品安全
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）