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
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产品关联
DOI
高影响因子
FOOD CHEMISTRY
·
2022
·
IF 9.231
Ying-yue Zhang,Mei-juan Zhao,Chun-yao Liu,Kai Ma,Tian-yu Liu,Fei Chen,Li-na Wu,De-jun Hu,Guang-ping Lv
Immunoassay-based techniques are important on-site screening tools for the detection of mycotoxins in cereals. This study aims to evaluate the trueness, precision, repeatability and cross-reactivity of commercially available test strips, ELISA kits and UHPLC-MS/MS on analyzing zearalenone, ochratoxin A, deoxynivalenol, T-2 toxin and fumonisin B 1 . The results showed that false negative rate (25.7 %–37.4 %) of all tested mycotoxins by test strips was higher than the false positive rate (0 %–31.0 %). The repeatability of ELISA kits at the declared LOD dispersed from −85.7 % to +98.4 %. ELISA kits were more accurate at 50 % of the maximum residue limit (MRL) of mycotoxins than 150 % and 200 %. All the tested deoxynivalenol/zearalenone derivatives had cross-reactivity with different level, and sample matrix could reinforce this overestimation of target mycotoxin. This study emphasized that higher-quality antibody screening and more analytical performance investigations are need to address for on-site detection of mycotoxins in the future.
研究领域:
食品科学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Gut Microbes
·
2022
·
IF 9.434
Liqiu Jia,Shufeng Weng,Jing Wu,Xiangxiang Tian,Yifan Zhang,Xuyang Wang,Jing Wang,Dongmei Yan,Wanhai Wang,Fang Fang,Zhaoqin Zhu,Chao Qiu,Wenhong Zhang,Ying Xu,Yanmin Wan
The origins of preexisting SARS-CoV-2 cross-reactive antibodies and their potential impacts on vaccine efficacy have not been fully clarified. In this study, we demonstrated that S2 was the prevailing target of the preexisting S protein cross-reactive antibodies in both healthy human and SPF mice. A dominant antibody epitope was identified on the connector domain of S2 (1147-SFKEELDKYFKNHT-1160, P144), which could be recognized by preexisting antibodies in both human and mouse. Through metagenomic sequencing and fecal bacteria transplant, we demonstrated that the generation of S2 cross-reactive antibodies was associated with commensal gut bacteria. Furthermore, six P144 reactive monoclonal antibodies were isolated from naïve SPF mice and were proven to cross-react with commensal gut bacteria collected from both human and mouse. A variety of cross-reactive microbial proteins were identified using LC-MS, of which E. coli derived HSP60 and HSP70 proteins were confirmed to be able to bind to one of the isolated monoclonal antibodies. Mice with high levels of preexisting S2 cross-reactive antibodies mounted higher S protein specific binding antibodies, especially against S2, after being immunized with a SARS-CoV-2 S DNA vaccine. Similarly, we found that levels of preexisting S2 and P144-specific antibodies correlated positively with RBD binding antibody titers after two doses of inactivated SARS-CoV-2 vaccination in human. Collectively, our study revealed an alternative origin of preexisting S2-targeted antibodies and disclosed a previously neglected aspect of the impact of gut microbiota on host anti-SARS-CoV-2 immunity.
研究领域:
免疫学
·
微生物学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2015
·
IF 0.986
Naifeng Xu,Liqiang Li,Shanshan Song,Liguang Xu,Hua Kuang,Chuanlai Xu
We developed a sensitive lateral flow immunoassay (LFI) for the detection of total malachite green (MG) residues in fish tissues. LFI was based on a colloidal gold nanoparticle-labeled monoclonal antibody against MG , which was generated by fast immunization of carboxymalachite green-bovine serum albumin. The half-maximum inhibition concentration (IC50) of MG monoclonal antibody (MG mAb) was 0.057 ng/ml with a linear range of 0.020–0.162 ng/ml. Following the optimization of LFI, a qualitatively visual limit of detection of 1 ng/ml and a semi-quantitatively limit of detection of 0.418 ng/ml (IC50 of the calibration curve) were obtained for total MG residues in 8 min. Cross-reactivity with most MG analogs was negligible, expect with crystal violet (CV; 78.6% cross-reactivity). LFI is a novel and sensitive assay based on MG mAb that has applications for the rapid detection of total MG and CV in fish products.
研究领域:
食品安全
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
FOOD CHEMISTRY
·
2022
·
IF 9.231
Yu Lu,Lujia Sun,Cheng Li,Xiaoqin Wang,Wenqing Li,Tong Zhao,Linjuan Huang,Zhongfu Wang
Extensive cross-allergic reactivity exists among food-borne plants. As most allergens are glycoproteins, protein sensitization has been extensively studied. However, little attention has been given to the glycan component of glycoproteins. Here, we qualitatively and quantitatively compared the N -glycans of eight cross-reactive plant glycoproteins derived from peach pollen, soy, pine nut, cashew nut, pistachio, walnut, almond, and hazelnut. Cross-reactive carbohydrate determinants (CCDs) were widely present in the allergic glycoproteins according to the following order: pollen > soybean > pistachios > pine nuts > walnuts > cashews > hazelnuts > almonds. The N -glycan structure presented clear differences between the germplasm and vegetative tissue. Importantly, fucosylation and xylosylation levels correlated positively with extensive cross-reactivity between plants, which suggests that CCDs may be one of the causes of cross-allergy. This result provides the foundation for mechanistic studies on plant-derived CCDs as a source of food cross-allergies.
研究领域:
生物化学
·
免疫学
✓ 已验证
DOI
高影响因子
FOOD CONTROL
·
2017
·
IF 3.496
Xiya Zhang,Meirong Song,Xuezhi Yu,Zhanhui Wang,Yuebin Ke,Haiyang Jiang,Jiancheng Li,Jianzhong Shen,Kai Wen
To reduce the incidence of false-positive and false-negative results caused by high or low cross-reactivity (CR%) values of the antibodies for total aflatoxins (AFs, AFB 1 +AFB 2 +AFG 1 +AFG 2 ) detection, a new broad-specific monoclonal antibody (MAb) with uniform affinity, named 5H3, was developed. Moreover, magnetic beads (MBs) replaced microplates as immobile phase to improve the sensitivity of the enzymatic immunoassay. Then, a direct competitive enzyme-linked immunosorbent assay (ELISA) based on MBs (MBs-dcELISA) that could simultaneously detect the total AFs with similar sensitivity was developed. Following optimization of conditions, the half maximal inhibitory concentrations (IC 50 ) of the MBs-dcELISA in buffer were 0.05 ng/mL for AFB 1 , 0.04 ng/mL for AFB 2 , 0.05 ng/mL for AFG 1 , 0.06 ng/mL for AFG 2 . The corresponding CR% values were 100%, 125%, 100% and 83.3%, respectively. The limit of detection (LOD) of the MBs-dcELISA for the total AFs was 0.21 ng/g with a working range from 0.22 ng/g to 19.8 ng/g, and the recoveries for the total AFs ranged from 74.5% to 96.5% with coefficients of variation (CV) under 12.1% in spiked maize samples. In addition, the MBs-dcELISA was more sensitive than the conventional dcELISA. Finally, the MBs-dcELISA was applied to screen 9 naturally contaminated maize samples and 6 spiked samples and the results indicated a good agreement with that obtain by HPLC-MS/MS method.
研究领域:
食品科学
·
免疫学
✓ 已验证
DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2017
·
IF 1.392
Xiujin Chen,Zhaozhou Li,Jinying Guo,Daomin Li,Hongli Gao,Yao Wang,Chuanlai Xu
A broad-specificity immunoassay was established to monitor marbofloxacin (MBF) and ofloxacin (OFX) residues in raw milk and porcine muscle samples. The main influential parameters in the assay buffer were investigated to improve the assay performance. The cross-reactivities (CRs) were lower than 0.25%, except for OFX (41.38%). The quantitative working ranges for MBF and OFX based on fortified matrices ranged from 0.11 to 30.85 ng mL−1, with a 50% inhibitory concentration of 0.76±0.12 ng mL−1 for MBF and 2.70±0.28 ng mL−1 for OFX. The detection limits of enzyme-linked immunosorbent assay (ELISA) ranged from 0.053 to 0.358 ng mL−1 in spiked samples. The mean recoveries were in the range of 80.52–95.46%, less than 12.06% of the coefficient of variation. A good correlation between the ELISA results and those from instrument methods demonstrated the practical application of the novel ELISA. Therefore, the immunoassay proposed is feasible for MBF and OFX analysis in animal-derived foods.
研究领域:
食品安全
·
免疫学
·
分析化学
✓ 已验证
DOI
高影响因子
Foods
·
2024
·Vol. 14
·期 1
·27
·
IF 4.7
Xingdong Yang,Yang Qu,Chenchen Wang,Lihua Wu,Xiaofei Hu
The ingestion of food contaminated with citrinin (CIT) poses a variety of health risks to humans and animals. The immunogens (CIT-COOH-BSA, CIT-H-BSA) and detection antigen (CIT-COOH-OVA, CIT-H-OVA) were synthesised using the active ester method (-COOH) and formaldehyde addition method (-H). A hybridoma cell line (3G5) that secretes anti-CIT monoclonal antibodies (mAbs) was screened via CIT-H-BSA immunisation of mice, cell fusion, and ELISA screening technology. The cell line was injected intraperitoneally to prepare ascites. The reaction conditions for the indirect competitive ELISA (ic-ELISA) were optimised, and anic-ELISA method for detecting CIT was preliminarily established. The results revealed that the IC50of CIT from optimisedic-ELISA was 37 pg/mL, the linear detection range was 5.9~230 pg/mL, and the cross-reaction (CR) rate with other analogues was less than 0.01%. The intra-assay and interassay sample recovery rates of CIT were 84.7~92.0% and 83.6~91.6%, and the coefficients of variation (CVs) were less than 10%. Theic-ELISA of CIT established in this study was not significantly different from the HPLC results and is rapid, highly sensitive and strongly specific, providing technical support for the detection of CIT.
研究领域:
食品安全检测
·
免疫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2018
·
IF 2.568
Min Feng,Steven Suryoprabowo,Hong Tao,Liqiang Liu,Qiankun Zheng,Hua Kuang
Clonidine (CLO) preferentially stimulates central α-2-adrenoceptors and causes both desirable and undesirable effects. Undesirable effects include sedation and withdrawal reactions, which occur as a sudden rise in arterial pressure, nervousness, agitation, and increased heart rate. An immunochromatographic test strip was developed for the rapid simultaneous detection of CLO and its cross-reactivity with apraclonidine (ACLO) in pig urine. The antigen CLO-OVA and goat anti-mouse IgG were attached to a nitrocellulose membrane as control line and test line. The cut-off limits of the test strips for CLO and ACLO were found to be 2.5 ng/mL in both 0.01 M PBS (pH 7.4) and pig urine. All the results were obtained within 5 min. The results revealed that the developed method is a sensitive, rapid, and simple tool for the detection of CLO and ACLO.
研究领域:
兽医学
·
分析化学
·
药理学
✓ 已验证
产品关联
DOI
高影响因子
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2014
·Vol. 21
·期 9
·5803-5809
·
IF 2.757
Li Ming,Sheng Enze,Yuan Yulong,Liu Xiaofeng,Hua Xiude,Wang Minghua
Europium (Eu 3+ )-labeled antibody was used as a fluorescent label to develop a highly sensitive time-resolved fluoroimmunoassay (TRFIA) for determination of clothianidin residues in agricultural samples. Toward this goal, the Eu 3+ -labeled polyclonal antibody and goat anti-rabbit antibody were prepared for developing and evaluating direct competitive TRFIA (dc-TRFIA) and indirect competitive TRFIA (ic-TRFIA). Under optimal conditions, the half-maximal inhibition concentration (IC 50 ) and the limit of detection (LOD, IC 10 ) of clothianidin were 9.20 and 0.0909 μg/L for the dc-TRFIA and 2.07 and 0.0220 μg/L for the ic-TRFIA, respectively. The ic-TRFIA has no obvious cross-reactivity with the analogues of clothianidin except for dinotefuran. The average recoveries of clothianidin from spiked water, soil, cabbage, and rice samples were estimated to range from 74.1 to 115.9 %, with relative standard deviations of 3.3 to 11.7 %. The results of TRFIA for the blind samples were largely consistent with gas chromatography ( R
2 = 0.9902). The optimized ic-TRFIA might become a sensitive and satisfactory analytical method for the quantitative monitoring of clothianidin residues in agricultural samples.
研究领域:
农业科学
·
分析化学
·
环境科学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL LETTERS
·
2023
·
IF 2
Peipei Cao,Xiangde Lai,Rui Zhang,Zhiling Chen,Bin Qiao,Lina Niu,Qiang Wu
Monkeypox, a rare zoonotic disease classified as a category B infectious disease, is caused by the monkeypox virus (MPXV), which has a broad host range. Since May 2022, human MPXV infection outbreak has spread to more than 100 countries, and China has reported 106 MPXV infections between June 2 and 30, 2023. Therefore, rapid and effective MPXV detection is essential to control and prevent its transmission. In this study, a rapid, cost-effective, and convenient fluorescent immunochromatographic assay (FICA) for MPXV antigen A35R detection was developed. Under the optimal reaction conditions, the visual detection limit of the proposed assay was 7.5 ng/mL and was completed within 15 min. There was no cross-reactivity with norovirus (NV) or dengue virus (DENV) controls. Additionally, the clinical application of the proposed method was demonstrated by testing human serum samples spiked with the MPXV antigen A35R. FICA was faster and more convenient than traditional enzyme-linked immunosorbent assays (ELISA) or polymerase chain reaction (PCR), and hence suitable for the early screening of MPXV infection. In summary, FICA could realize rapid, low-cost, sensitive, and specific MPXV detection hold potential application prospects in clinical settings.
研究领域:
病毒学
·
传染病
·
分子诊断
·
临床微生物学
✓ 已验证
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2019
·
IF 9.518
Yun Zhang,Wenqing Tan,Yang Zhang,Huili Mao,Shuyou Shi,Liangwei Duan,Hui Wang,Junping Yu
To develop a specific method for the detection of S. aureus , chicken anti-protein A IgY was adopted for specifically capturing S. aureus , depending on the specific recognition of staphylococcal protein A (SPA) by chicken anti-protein A IgY, which can eliminate the interference from protein G-producing Streptococcus . HRP labeled IgG , Fc region of which has a high affinity towards SPA, was paired with IgY for the colorimeter analysis of the system. By optimizing the system, a super-low detection limit of 11 CFU of S. aureus in 100 μL PBS without enrichment, with a linear range from 5.0 × 10 2 CFU mL −1 to 5.0 × 10 4 CFU mL −1 was obtained. The entire assay was accomplished in less than 90 min and no cross-reactivity with the other tested bacterial species was observed. Moreover, the developed assay has been applied for the detection of S. aureus in three different types of real samples (sodium chloride injection, apple juice and human urine) with satisfactory results. To the best of our knowledge, it is the first time to report using chicken anti-protein A IgY and any IgG to detect S. aureus based on the dual-recognition mode of SPA. The novel method opened up a way for monitoring S. aureus in food samples with high sensitivity, specificity and simple operation.
研究领域:
微生物学
·
分析化学
·
食品安全
✓ 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）