Survey of Deoxynivalenol Contamination in Agricultural Products in the Chinese Market Using An ELISA Kit

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−1 limit of detection (LOD) in working buffer and a 200 ng g−1 LOD 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.

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Determination of Aflatoxin M1 in Raw Milk Using an HPLC-FL Method in Comparison with Commercial ELISA Kits—Application in Raw Milk Samples from Various Regions of Greece

Veterinary Sciences ◽

10.3390/vetsci8030046 ◽

2021 ◽

Vol 8 (3) ◽

pp. 46

Author(s):

Martha Maggira ◽

Maria Ioannidou ◽

Ioannis Sakaridis ◽

Georgios Samouris

Keyword(s):

Limit Of Detection ◽

Reference Method ◽

Raw Milk ◽

Aflatoxin M1 ◽

Enzyme Linked Immunosorbent Assay ◽

Fluorescence Detector ◽

Elisa Kit ◽

Milk Samples ◽

Of Greece

The highly toxic Aflatoxin M1 (AFM1) is most often detected in milk using an Enzyme-Linked-Immunosorbent Assay (ELISA) for screening purposes, while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-FL) is the reference method used for confirmation. The aim of the present study was the comparison between three commercially available ELISA kits and a newly developed HPLC-FL method for the determination of the AFM1 in milk samples. The developed HPLC-FL method was validated for the AFM1 and Aflatoxin M2 (AFM2), determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results. All three ELISA kits were also validated and showed equally good performance with high recovery rates. Moreover, the Limit Of Detection (LOD) and Limit Of Quantification (LOQ) values were found to be significantly lower than the Maximum Residue Limit (MRL) (50 ng kg−1). After the evaluation of all three commercial kits, the ELISA kit with the optimum performance along with the HPLC method was used for the determination of AFM1 in raw cow’s, goat’s, and sheep’s milk samples (396) obtained from producers in different regions of Greece. The evaluation of both methods showed that this ELISA kit could be considered as a faster and equally reliable alternative method to HPLC in routine analysis for the determination of AFM1 in milk.

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Pitfalls in the Immunochemical Determination of β‑Lactam Antibiotics in Water

Antibiotics ◽

10.3390/antibiotics10030298 ◽

2021 ◽

Vol 10 (3) ◽

pp. 298

Author(s):

Alexander Ecke ◽

Rudolf J. Schneider

Keyword(s):

Limit Of Detection ◽

Cost Effective ◽

Enzyme Linked Immunosorbent Assay ◽

Degree Of Hydrolysis ◽

Site Analysis ◽

Antibody Recognition ◽

Immunochemical Determination ◽

Key Factor ◽

Lactam Ring

Contamination of waters with pharmaceuticals is an alarming problem as it may support the evolution of antimicrobial resistance. Therefore, fast and cost-effective analytical methods for potential on-site analysis are desired in order to control the water quality and assure the safety of its use as a source of drinking water. Antibody-based methods, such as the enzyme-linked immunosorbent assay (ELISA), can be helpful in this regard but can also have certain pitfalls in store, depending on the analyte. As shown here for the class of β-lactam antibiotics, hydrolysis of the β‑lactam ring is a key factor in the immunochemical analysis as it influences antibody recognition. With the antibody used in this study, the limit of detection (LOD) in the immunoassay could be significantly reduced by hydrolysis for the five tested penicillins, with the lowest LOD for carbenicillin (0.2 nmol/L) and the greatest impact on penicillins G and V (reduction by 85%). In addition to enhanced quantification, our strategy also provides access to information about the degree of hydrolysis in water samples as shown for the most abundant penicillin amoxicillin.

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Characterization and validation of fragment antigen-binding (Fab) antibody-based immunoassay for deoxymiroestrol, a potent phytoestrogen from Pueraria candollei

10.22541/au.161270515.50311593/v1 ◽

2021 ◽

Author(s):

Worapol Sae-foo ◽

Supaluk Krittanai ◽

Wipawee Juengsanguanpornsuk ◽

Gorawit Yusakul ◽

Tharita Kitisripanya ◽

...

Keyword(s):

Quantitative Method ◽

Estrogenic Activity ◽

Limit Of Detection ◽

Hormone Replacement ◽

Enzyme Linked Immunosorbent Assay ◽

Antigen Binding ◽

Pueraria Candollei ◽

Specific Determination ◽

Fragment Antigen Binding

Deoxymiroestrol is the most potent phytoestrogen in chromenes group that has been found in Pueraria candollei, Thai name known as Kwao Krua Khao. Several studies reported estrogenic activity of P. candollei in order to using as hormone replacement therapy for postmenopausal women. Previously, specific determination of deoxymiroestrol content by enzyme-linked immunosorbent assay (ELISA) using polyclonal antibody (pAb) have been reported. However, production of pAb has limitation and variability from different batches. Therefore, in this study, we established quantitative method for determination of deoxymiroestrol using fragment antigen-binding (Fab) antibody-based immunoassay. The developed immunoassay has specificity to deoxymiroestrol with a calibration range of 15.6-1000 ng mL-1. Precision including intra-assay and inter-assay are 1.48-7.11 and 0.58-9.31%, respectively. Accuracy of the assay showed in recovery between 99.77-101.61% when spike deoxymiroestrol standard into the samples. The limit of detection (LOD) is 30.80 ng mL-1. Comparation antibody-based immunoassay for determination of deoxymiroestrol using Fab with pAb was represented consistency (R2 = 0.9807) when analysis roots bark of Pueraria candollei from difference areas. Therefore, this development assay can apply to determine deoxymiroestrol content in the plant samples.

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Evaluation of Performance of a Commercial Monoclonal Antibody–Based Fenitrothion Immunoassay and Application to Residual Analysis in Fruit Samples

Journal of Food Protection ◽

10.4315/0362-028x-69.1.191 ◽

2006 ◽

Vol 69 (1) ◽

pp. 191-198 ◽

Cited By ~ 7

Author(s):

EIKI WATANABE ◽

KOJI BABA ◽

HEESOO EUN ◽

TOMOHITO ARAO ◽

YASUO ISHII ◽

...

Keyword(s):

Monoclonal Antibody ◽

Dynamic Range ◽

Limit Of Detection ◽

Close Correlation ◽

Gas Chromatography Mass Spectrometry ◽

Enzyme Linked Immunosorbent Assay ◽

High Recovery ◽

Elisa Kit ◽

Fruit Samples ◽

Standard Solutions

The performance of a commercially available enzyme-linked immunosorbent assay (ELISA) kit containing highly sensitive monoclonal antibodies against fenitrothion was assessed. The experimentally estimated dynamic range (0.087 to 2 ng/g) agreed with that established by the kit manufacturer (0.075 to 1 ng/g). The linearity of the standard curve produced for the kit-assembled standard solutions (slope = −0.3829) agreed with that of the curve produced for the self-made standard solutions (slope =−0.3619). The sensitivity (I50 value) and the limit of detection for the kit were 0.23 and 0.033 ng/g, respectively. Selectivity of the ELISA indicates that the monoclonal antibody can readily distinguish the target pesticide from other structurally related analogs and some metabolites (oxon forms), with the exception of ethyl O-(p-nitrophenyl) phenyl phosphonothionate (EPN), parathion-methyl, and parathion. Methanol was the best organic solvent for the kit, with optimal sensitivity observed at a final concentration in the well of 10% (vol/vol) or less. Matrix interferences were minimized by direct dilution with water (60-fold) of the methanolic extracts from apple and peach samples. To extract fenitrothion from these two agricultural products as simply and rapidly as possible, three extraction methods were used. The extraction method that involved shaking by hand for 3 min was the best among the three methods. High recovery percentages (116.6% for apple and 110.8% for peach) were obtained. Validation of the ELISA method was carried out using the gas chromatography–mass spectrometry method, resulting in high recovery and close correlation of results (r > 0.95). These findings strongly suggest that the ELISA kit may be routinely employed for on-site fenitrothion screening of fruit samples.

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Determination of Prolactin in Canine Saliva: Is it Possible to Use a Commercial ELISA kit?

Animals ◽

10.3390/ani9070418 ◽

2019 ◽

Vol 9 (7) ◽

pp. 418 ◽

Cited By ~ 1

Author(s):

Gutiérrez ◽

Gazzano ◽

Torracca ◽

Meucci ◽

Mariti

Keyword(s):

Stress Response ◽

Immunosorbent Assay ◽

Matrix Effect ◽

Enzyme Linked Immunosorbent Assay ◽

Blood Samples ◽

Elisa Kit ◽

Cases Study ◽

Dog Blood

Prolactin has been reported to be a remarkable index of stress response, both acute and chronic, in several species. The use of biological matrixes other than blood is receiving increasing interest in the study of hormones, due to the lower invasiveness in collection. This research aimed to investigate the possibility of using a commercial ELISA (enzyme-linked immunosorbent assay) kit for measuring canine prolactin in blood for the quantification of canine prolactin in saliva. Study 1 consisted of a validation protocol, using saliva samples collected from lactating and non-lactating dogs. Study 2 was conducted to investigate a possible correlation between prolactin concentration in saliva and plasma in sheltered dogs by using the same kit. Prolactin values were reliably read only when they came from blood samples, not from saliva, but tended to be low in most of the cases. Study 1 showed that saliva had a matrix effect. In study 2, saliva prolactin levels were low and in 42.9% of cases, not readable. No correlation between prolactin values in plasma and saliva was found (ρ=0.482; p=0.274). These findings suggested that the determination of prolactin in dog saliva through an ELISA kit created for measuring prolactin in dog blood was unreliable.

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A Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Determination of Homoharringtonine

Planta Medica ◽

10.1055/a-0578-8689 ◽

2018 ◽

Vol 84 (14) ◽

pp. 1038-1044 ◽

Cited By ~ 3

Author(s):

Benyakan Pongkitwitoon ◽

Seiichi Sakamoto ◽

Rika Nagamitsu ◽

Waraporn Putalun ◽

Hiroyuki Tanaka ◽

...

Keyword(s):

Monoclonal Antibody ◽

Immunosorbent Assay ◽

Myeloid Leukemia ◽

High Performance ◽

Sodium Periodate ◽

Limit Of Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Study Support ◽

Mediated Oxidation

AbstractHomoharringtonine (HHT), also known as omacetaxine, is a natural compound found in the genus Cephalotaxus and is a promising pharmaceutical drug used for the treatment of chronic or accelerated phase chronic myeloid leukemia. As a tool for the quantitative determination of HHT, a specific monoclonal antibody against HHT (MAb 6A1) was generated by conjugates prepared via sodium periodate-mediated oxidation. The developed indirect competitive enzyme-linked immunosorbent assay (icELISA) using MAb 6A1 was found to be highly specific and sensitive with a limit of detection for HHT of 48.8 ng/mL. Validation assays to evaluate precision and accuracy of the method were conducted by the use of intra- and inter-assay analysis, recovery test, and comparison analysis between the amounts of HHT determined by ELISA and high-performance liquid chromatography. These results revealed that the established icELISA using MAb 6A1 is specific, sensitive, and reliable enough to be applied to the qualitative analysis for HHT. Furthermore, the results of this study support the usefulness of sodium periodate as a reagent for the conjugation between Cephalotaxus alkaloids and proteins for producing specific antibodies.

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Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody

Planta Medica ◽

10.1055/s-0043-102689 ◽

2017 ◽

Vol 83 (10) ◽

pp. 855-861 ◽

Cited By ~ 7

Author(s):

Tharita Kitisripanya ◽

Supaluk Krittanai ◽

Orapin Udomsin ◽

Kamonthip Jutathis ◽

Jukrapun Komaikul ◽

...

Keyword(s):

Monoclonal Antibody ◽

Estrogenic Activity ◽

Limit Of Detection ◽

Simple Procedure ◽

Enzyme Linked Immunosorbent Assay ◽

Efficacy And Safety ◽

Limit Of Quantitation ◽

Estrogenic Action ◽

White Kwao Krua

AbstractMiroestrol is a chromene with potent estrogenic activity present in Pueraria candollei, commonly known as White Kwao Krua. Although this compound is only present in low amounts in the plant, it plays an important role in the estrogenic action of P. candollei products. As a tool for further studies about the efficacy and safety of P. candollei as a phytoestrogenic supplement, we generated a novel monoclonal antibody against miroestrol. This anti-miroestrol monoclonal antibody was used to develop an immunoassay for the determination of miroestrol content, which can be used for quality control purposes of P. candollei. The developed ELISA against miroestrol has a calibration range of 10–780 ng/mL miroestrol, a limit of detection of 3.5 ng/mL, and a limit of quantitation of 12.2 ng/mL. According to the validation analysis, the established ELISA is precise, accurate, specific, and sensitive for miroestrol detection in plants. Furthermore, the anti-miroestrol monoclonal antibody was used to prepare an immunoaffinity column for the isolation of miroestrol from the tuberous root of P. candollei. The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel.

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Single-Laboratory Validation of the Biosense Direct Competitive Enzyme-Linked Immunosorbent Assay (ELISA) for Determination of Domoic Acid Toxins in Shellfish

Journal of AOAC International ◽

10.1093/jaoac/90.4.1000 ◽

2007 ◽

Vol 90 (4) ◽

pp. 1000-1010 ◽

Cited By ~ 9

Author(s):

Hans Kleivdal ◽

Sven-Inge Kristiansen ◽

Mona V Nilsen ◽

Lyn Briggs

Keyword(s):

Immunosorbent Assay ◽

Domoic Acid ◽

Matrix Effects ◽

Limit Of Detection ◽

Method Comparison ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Shellfish Poisoning ◽

Relative Standard

Abstract Method validation was conducted for an enzyme-linked immunosorbent assay (ELISA) for the determination of domoic acid (DA) toxins, known to give amnesic shellfish poisoning (ASP) symptoms, in shellfish. The calibration curve range of the assay is approximately 10260 pg/mL, with a dynamic working range for DA toxins in shellfish from 0.01 to at least 250 mg/kg. The ASP ELISA showed no significant cross-reactivity to structural analogs, and proved to be robust to deliberate alterations of the optimal running conditions. The shellfish matrix effects observed with mussels, oysters, and scallops were eliminated by diluting shellfish extracts 1:200 prior to analysis, leading to a limit of detection at 0.003 mg/kg. Thirteen blank shellfish homogenates were spiked with certified mussel material containing DA to levels in the range of 0.125 mg DA/kg, and analyzed in quadruplicate on 3 different days. The relative standard deviation (RSD) under intra-assay repeatability conditions ranged from 6.5 to 13.1%, and under interassay repeatability conditions the RSD ranged from 5.7 to 13.4%, with a mean value of 9.3%. The recoveries ranged from 85.5 to 106.6%, with a mean recovery of 102.2%. A method comparison was conducted with liquid chromatography with ultraviolet detection, using naturally contaminated scallop samples (n = 27) with DA levels at 0244 mg/kg. The overall correlation coefficient was 0.960 and the slope of the regression was 1.218, indicating a good agreement between the methods.

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Detection of bisphenol A in water samples using ELISA determination method

Water Science & Technology Water Supply ◽

10.2166/ws.2011.008 ◽

2011 ◽

Vol 11 (1) ◽

pp. 55-60 ◽

Cited By ~ 4

Author(s):

J. Zheng ◽

S. Q. Zhao ◽

X. T. Xu ◽

K. Zhang

Keyword(s):

Drinking Water ◽

Bisphenol A ◽

Water Samples ◽

Limit Of Detection ◽

Industrial Effluent ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Uv Spectrophotometry ◽

Inhibition Concentration

In order to study whether bisphenol A (BPA) can pass into drinking water from polycarbonate barrel and exist in the river and industrial effluent the indirect competitive enzyme-linked immunosorbent assay (ELISA) for the determination of BPA was established. The results presented an inhibition concentration at 50% absorbance (IC50) of 0.123 mg L−1, and the limit of detection (LOD) is 9.934 μg L−1. The specificity of antiserum was proved well because the cross-reactivity with benzene, tert-butylbenzene, hydroquinone and o-hydroxybenzoic acid were found lower than 0.01%, except phenol was 0.26%. The method was found to be reliable and repeatable. It was used for monitoring the concentration of BPA in the barreled drinking water. The results confirmed BPA can pass into barreled drinking water from the polycarbonate barrel and concentration increased as days went on. A certain content of BPA was found in industrial effluent. The results of ELISA were consistent with the results of UV spectrophotometry. BPA could not be found in the water samples obtained from Zhujiang River. The established method shows specific recognition of BPA and could be applied in detection of environmental BPA.

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Usefulness of ultrasensitive anti-Müllerian hormone assay to predict follicle growth in patients with primary ovarian insufficiency

10.21203/rs.2.22083/v1 ◽

2020 ◽

Author(s):

Yukiyo Kasahara ◽

Satoko Osuka ◽

Bayasula Bayasula ◽

Natsuki Nakanishi ◽

Tomohiko Murase ◽

...

Keyword(s):

Limit Of Detection ◽

Early Stage ◽

Primary Ovarian Insufficiency ◽

Enzyme Linked Immunosorbent Assay ◽

Follicle Growth ◽

Ovarian Insufficiency ◽

Elisa Kit ◽

Median Serum ◽

Positive Serum ◽

Low Levels

Abstract Background: Patients with primary ovarian insufficiency (POI) present with follicle growth occasionally, however, it is difficult to predict the exact cycles with follicle growth. Serum anti-Müllerian hormone (AMH), a useful marker for ovarian reserve, is produced in early-stage follicles. Therefore, AMH levels should reflect the existence of small follicles which are difficult to detect using ultrasonography and may be useful for predicting follicle growth in patients with POI. Methods: Using an ultrasensitive enzyme-linked immunosorbent assay (ELISA) kit, we found very low serum AMH levels of patients with POI; we further assessed the follicle growth of each patient and their cycles to clarify the usefulness of measuring serum AMH levels for predicting the follicle growth in patients with POI. Results: A total of 91 cycles of 19 patients with POI were analyzed in this study. Five patients presented with positive serum AMH levels during the observational periods and all five experienced follicle growth. Only two of the 14 patients with negative serum AMH levels had follicle growth. Serum AMH and follicle-stimulating hormone (FSH) levels in cycles with follicle growth were significantly higher than in cycles without follicle growth. The median serum AMH level (2.77 pg/mL; 25th, 75th percentiles: 0.0, 9.64) in cycles with follicle growth were lower than the lower limit of detection of conventional AMH ELISA kits. The positive predictive value of positive serum AMH levels for follicle growth was higher than that of FSH (<10 mIU/mL).Conclusions: Measuring very low levels of serum AMH using picoAMH assays is useful for the prediction of follicle growth in patients with POI.

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