Food Analytical Methods
·
2014
·Vol. 7
·期 10
·2163-2168
·
IF 1.802
Suryoprabowo Steven,Liu Liqiang,Peng Juan,Kuang Hua,Xu Chuanlai
Fast and sensitive screening of antibiotics is a great need in food quality assurance. A monoclonal antibody specific to fluoroquinolones (FQ) was obtained using ciprofloxacin (CPF) derivant as hapten. The antibody was characterized with broad recognition to CPF (100 %), enrofloxacin (ENF, 73.89 %), norfloxacin (73.57 %), nadifloxacin (67.28 %), danofloxacin (53.09 %), pefloxacin (50.26 %), lomefloxacin (35.66 %), enoxacin (12.40 %), and sarafloxacin (3.23 %). Then, an enzyme linked immunosorbent assay (ELISA) was established. The optimized concentrations of coating antigen and antibody were 0.1 and 0.5 μg/ml, respectively. Various parameters including pH values, ionic strength and the concentration of antibody and antigen were optimized for this assay. The ELISA was found to have the best sensitivity in assay buffer of pH 6 and sodium chloride content 1.6 % (m/v) using CPF as tested compound. Fortified milk samples with CPF and ENF (50 and 250 μg/l) and fortified chicken samples (10 and 50 μg/kg) were analyzed with the proposed measure. The ELISA was examined with recovery range of 94–104 % for milk detection and 93–108 % for chicken detection. The coefficient variation data for intra-assay and inter-assay ranged from 4.24 % to 12.16 %. All results indicate the potential extensive application prospects of this ELISA in FQ monitoring for food safety.
研究领域:
分析化学
·
食品科学
·
免疫学
✓ 已验证
产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2022
·
IF 5.304
Jie Liu,Aihong Wu,Shanshan Song,Liguang Xu,Liqiang Liu,Chuanlai Xu,Hua Kuang
Pendimethalin (PDM), a dinitroaniline group herbicide, is extensively used for controlling weeds in amounts of agricultural crops. In this work, Hapten-1 and Hapten-2 were synthesized for the preparation of an anti-PDM monoclonal antibody (mAb), which was used to establish an indirect competitive ELISA. Based on the anti-PDM mAb, we developed an immunochromatographic assay (ICA) for detection of PDM in potato and apple. Under optimal conditions, the ICA exhibited a cut-off value of 200 ng/g and the visible limit of detection was 10 ng/g for apple and potato samples in qualitative detection. Analysis of quantitative detection suggested that the limit of detection were 5.4 and 3.2 ng/g in apple and potato samples, respectively. Moreover, the results obtained from the developed ICA strip were shown to coincide with those of LC–MS/MS in the analysis of apple and potato samples. In addition, the ICA exhibited excellent performance with regard to sensitivity, speed, and portability in contrast to previous determination methods for PDM. Hence, the proposed ICA strip showed superior prospects in PDM residue determination.
研究领域:
农业科学
·
分析化学
·
免疫学
✓ 已验证
DOI
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2014
·
IF 2.829
Yajing Lei,Xiaoyi Li,Muhammad Sajid Hamid Akash,Lifang Zhou,Xiaojing Tang,Weixing Shi,Zhiming Liu,Shuqing Chen
Salbutamol (SAL) is known as quick-acting β-2 receptor agonist and its use in humans for pulmonary diseases and/or in animal feed is limited because of associated potential hazardous effects on health. Several techniques are available for the detection of SAL, but are expensive and time consuming. Here for the first time, a novel pre-assembled DNA immunoprobe-based immuno-PCR assay was developed to investigate the levels of SAL in human urine samples and compared the proposed immuno-PCR assay for the detection of SAL with that of direct competitive ELISA. Under the optimized conditions, the assay showed a linear range over seven orders of magnitude, whereas the limit of detection of SAL in buffer was found to be 21 fg/mL. Current immuno-PCR assay exhibited a 300-fold better detection limit for SAL as compared to one achieved with direct competitive ELISA using the same antibody. In conclusion, it has been found that the developed method is highly sensitive and simple method that can significantly enhance the detection sensitivity for small molecules. Moreover it can be modified and used for detecting other small molecules and/or chemicals that exist in the environment and are responsible for the environmental pollution.
研究领域:
分析化学
·
分子生物学
·
环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Applied Sciences-Basel
·
2025
·Vol. 15
·期 7
·
IF 2.5
Tianyu Hu,Bingying Liu,Siqian He,Yuanjie Teng,Zaifa Pan
Insulin-like growth factor-1 (IGF-1) is a regulatory factor closely associated with diabetes, obesity, and breast cancer, and it also acts as one of the most abundant growth factors in bovine colostrum. Current methods generally have the problem of low sensitivity, a time-consuming nature, and low stability, which makes it difficult to crack down on the false advertising of IGF-1 content in dairy products. In this work, an ultrasensitive fluorescent enzyme-linked immunosorbent assay (ELISA) is proposed, where the antibody and the target are combined in the form of a “sandwich” to ensure the accuracy and specificity of the assay. IGF-1 is quantified based on an effective hydrogen peroxide (H2O2) probe with 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) as the fluorogenic substrate. The proposed fluorescent sandwich ELISA has a low limit of detection (LOD) of 77.29 pg/mL, fast experimental process within 1 h, and stable signal of 1 h. Furthermore, multi-step pretreatment methods for bovine colostrum powders are established to remove interfering substances, including fat, casein, and binding proteins, achieving the accurate and specific detection of IGF-1. IGF-1 recovery studies on treated bovine colostrum powders exhibit good recovery rates ranging from 91.71% to 102.32%, which proves the feasibility of detecting IGF-1 in real bovine colostrum.
研究领域:
分析化学
·
食品科学与技术
·
生物化学
·
免疫学
·
临床诊断
✓ 已验证
产品关联
DOI
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.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYST
·
2023
·Vol. 149
·期 2
·442-450
·
IF 4.2
Ting Wang,Ting Zhou,Kang Wu,Junlin Cao,Yuze Feng,Jianguo Li,Anping Deng
Zearalenone (ZEN) is one of the most toxic mycotoxins widely found in agricultural products. In this study, a sensitive enzyme-linked immunosorbent assay (ELISA) integrated with immunoaffinity column extraction for the detection of ZEN in food and feed samples was developed. A ZEN derivative containing a carboxylic group was first synthesized and then linked to bovine serum albumin (BSA). The formed ZEN–BSA conjugate was used as the immunogen for the production of the monoclonal antibody (mAb) against ZEN. The hybridoma clones (1G5) capable of secreting antibodies against ZEN were successfully selected. Based on this mAb, the IC50 and LOD of the ELISA for ZEN were 0.37 ng mL−1 and 0.04 ng mL−1, respectively, which were 1.6–308.1 times lower than those in the published ELISAs, indicating the high sensitivity of our assay. There was no cross-reactivity of the mAb with other four mycotoxins (patulin, AFB1, DON, and OTA). Due to the high similarity in molecular structures among ZEN and its homologs (α-zearalanol, β-zearalanol, zearalanone, α-zearalenol, β-zearalenol), the CR values of the mAb with the homologs were within 3.59%–105.71%. Taking advantage of plenty of mAb, the immunoaffinity column was prepared by immobilizing the mAb on Sepharose-4B gel and filling it into an SPE column. ZEN spiked samples (corn, wheat, feed) were extracted using an immunoaffinity column and measured by ELISA and HPLC-FLD simultaneously. The recoveries of the ELISA for ZEN in the spiked samples were 92.46–105.48% with RSDs of 4.87–10.11%. A good correlation between ELISA (x) and HPLC-FLD (y) with the linear regression equation y = 1.0589x + 1.43815 (R2 = 0.998, n = 6) was obtained. To verify the applicability, the proposed ELISA was also applied to some real samples randomly collected from a local market. It was proven that the newly produced mAb-based ELISA was a feasible and sensitive method for the detection of ZEN in food and feed samples.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Guojun Weng,Xun Shen,Jianjun Li,Jingyuan Wang,Jian Zhu,Junwu Zhao
The etching of gold nanorods/nanobipyramid, or silver-coated nanorods/nanobipyramid inducing plasmon changes represents an efficient strategy to improve the performance of enzyme-linked immunosorbent assay (ELISA). However, the effect of shape on the sensitivity was negligible, especially the thickness of coated silver shell. Here, we propose a plasmonic ELISA for multi-colorimetric detection of CRP based on the etching of Ag-coated Au nanobipyramid (Au [email protected] ). The effect of silver shell thickness on the sensitivity of plasmon peak shifting was investigated by experiments and DDA calculations. The relationship between the Ag shell thickness and the sensitivity of plasmon peak shifting was obtained. Our results reveal that the thickness of coated Ag shell acts as a key factor in the multi-color change of Au [email protected] etching. It is found that Au [email protected] with medium Ag shell thickness and rod-like shape has the higher sensitivity and is suitable for sensing. At the optimized most sensitive Ag shell, the detection limit of proposed plasmonic ELISA for CRP was determined to be 0.09 ng/mL with a spectrometer in the range from 0.09 ng/mL to 25 ng/mL. Importantly, the visual detection limit was 0.78 ng/mL, which allows the differential diagnosis with the naked eye. Compared with traditional ELISA with the monochromatic intensity variations, the multi-color ELISA proposed in this study has a large linear range and rich color variation for high-sensitivity and naked-eye semi-quantitative detection.
✓ 已验证
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
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 11
·1-12
·
IF 5.3
Nie Zheng,Sun Juan,Wu Fangjun,Lai Fujie,Yang Xudong,Chen Shaoyang,Wang Yang,Jiang Yong
A novel immunomagnetic bead was prepared and designed to be combined with a smartphone app for the rapid detection of SARS-CoV-2 antibody in serum. The nucleocapsid protein (N protein) of SARS-CoV-2 was immobilized on the prepared magnetic polymer microspheres (Fe 3 O 4 @PMAA) with core-shell structure by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimid/N-hydroxy succinimide (EDC/NHS) to form the immunomagnetic beads (Fe 3 O 4 @PMAA-NP) capable of binding antibodies specifically. Combining its magnetic separability with the image analysis function of the smartphone app, a novel detection platform was established on a 96-well plate. The platform can realize rapid separation and colorimetric detection of antibodies within 30 min, and the detection limit is 0.5 µg/mL. This method has the advantages of portability, rapidity, high sensitivity and low cost, which is suitable for home self-test and on-site detection. This study not only provides an innovative method for the antibody detection of SARS-CoV-2 in serum, but also provides a favorable reference for the practical application of functionalized magnetic polymer materials in the medical field. Graphical
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Liang Luo,Yingjie Zhang,Xiangjun Zhao,Weilin Wu,Jie Fei,Xuezhi Yu,Kai Wen,Jianzhong Shen,Yantong Pan,Zhanhui Wang
Xylazine (XYL) is an illicit adulterant in opioids and an approved veterinary sedative drug, which has been abused, misused, and residued in food samples, endangering people health, causing drug-facilitated crimes and even death. Immunoassay used antibody as core biomaterial could to achieve highly sensitive and rapid detection screening purpose for XYL in situ . Here, we rationally designed four novel XYL haptens with different spacer arms to produce antibodies with high affinity and specificity. Ten monoclonal antibodies (mAbs) were obtained and mAb 7H5 showed a high affinity with IC 50 of 0.23 ng mL −1 and ignorable cross-reactivity for the other eight analogs. One highly sensitive indirect competitive ELISA (icELISA) and lateral flow immunoassay (LFIA) were established based on heterologous haptens for XYL detection in a series of human urine and food samples. The limit of detections (LODs) of the icELISA were 0.10–2.24 μg L −1 with recovery of 83.5 % to 128.6 % and CV below 15.8 % in different samples, while, the visual limits of detection (vLOD) of LFIA were 0.10–1.80 μg L −1 with the cut-off value of 0.60–4.80 μg L −1 . In addition, the molecular recognition mechanism of mAbs was explored. The study provides powerful tools for rapid screening of XYL in human urine and food samples for the first time.
研究领域:
免疫学
·
药物检测
·
食品安全
✓ 已验证
产品关联
DOI
PubMed
高影响因子
International Journal of Electrochemical Science
·
2024
·
IF 1.3
Tao Zhong,Zuhua Jiang,Yuxuan Xing,Sicong Jiang,Yuan Wu,Shiyuan Yao
This study presents the development of an electrochemical immunosensor utilizing gold nanoparticles for the detection of creatine kinase (CK), a crucial cardiac marker. Gold nanoparticles with an average diameter of 18.3 ± 2.1 nm were synthesized and characterized, exhibiting a characteristic surface plasmon resonance peak at 520 nm. The construction of the immunosensor was observed through cyclic voltammetry and electrochemical impedance spectroscopy, showing notable alterations in electron transfer rates during each step of the assembly process. Optimal performance was achieved at pH 7.4, with an antibody concentration of 100 μg/mL and an incubation time of 45 minutes. The sensor demonstrated a sensitivity of 152.6 Ω/(ng/mL) and maintained its response in the presence of common interfering substances, with maximum signal deviation of 7.2 %. Testing with spiked human serum samples yielded recovery rates between 96.5 % and 103.8 %. Comparison with ELISA using clinical samples further validated the sensor's accuracy. This AuNP-based immunosensor offers a promising platform for rapid and sensitive CK detection in cardiac health assessment.
研究领域:
分析化学
·
生物传感器
·
心血管疾病诊断
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2026
·
IF 6.1
Jinlong Yuan,Han Sun,Yuhan Chen,Haohan Zhang,Wanjian Liu,Mingyang Wang,Zhonghuang Yang,Aihua Liu
Solid phase panning combined with liquid phase panning to screen specific peptide. • Specific peptide (GS24) binding to RSV GP with good affinity obtained. • Peptide GS24a-based sensitive competitive magnetic bead ELISA for RSV GP developed. • Clinical samples detected, agreeing well with results of test strips. Respiratory syncytial virus (RSV) is the leading cause of severe lower respiratory tract infections in infants, young children and immunocompromised individuals, to which effective vaccines and specific antiviral therapies remain elusive. Although several RSV vaccines and long-acting monoclonal antibodies have been approved for specific populations, rapid and sensitive diagnostic tools are still critical for early clinical triage. RSV glycoprotein (GP) is a highly glycosylated adhesion protein in mediating viral adhesion and immune escape, making it an ideal target for antigen detection. Herein a combination of solid phase panning with liquid phase panning strategy was explored to isolate peptide GS24 (sequence: SRTCTPNFCYLG) with high affinity and specificity to RSV GP. Molecular dynamics simulation indicates that Arg 2 and Phe 8 residues on GS24 play a crucial role in binding, due to lower binding energies. Glu232 and Glu284 residues on the RSV GP interact with Arg 2 of the peptide, respectively, to form two pairs of hydrogen bonds (Glu232-Arg2, Glu284-Arg2). Additionally, there exists π-π stacking interaction between Pro234 of RSV GP and Phe8 of the peptide. Then the peptide GS24a was used as a recognition probe, and coordination reaction between L-ascorbic-acid 2-phosphate and Fe 3+ as colorimetric signal, a signal-enhanced competitive binding of peptide/alkaline phosphatase-conjugated streptavidin complex to magnetic bead-immobilized antigen was constructed. The as-developed signal-enhanced competitive ELISA can detect target RSV GP within the linear range of 0.15-10 ng/mL and detection limit of 0.1 ng/mL. Finally, the proposed method was applied to detect human nasopharyngeal swab samples reproducibly, with the results in agreement well with commercial test strip, suggesting its good accuracy. This proposed method features simplicity, convenience and good sensitivity, which may offer a promising alternative for point-of-care RSV diagnosis. This work also provides guidance for developing similar immunoassay to detect other respiratory diseases. Download: Download high-res image (281KB) Download: Download full-size image
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）