scholarly journals Enzyme-linked immunosorbent assay (ELISA) for the screening of dioxins in fish samples

2021 ◽  
Author(s):  
Elaine Yu-Lan Chen
Keyword(s):  
Immunosorbent Assay ◽  
High Resolution Mass Spectrometry ◽  
Gel Permeation Chromatography ◽  
The Other ◽  
Alternative Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fish Sample ◽  
Purification Method ◽  
Screening Technique ◽  
Fish Samples

Dioxins are environmental contaminants that are toxic to humans. The conventional analytical method for dioxins, gas chromatography - high resolution mass spectrometry, is extremely time-consuming and expensive. Research is needed to find alternative methods that will increase sample throughput while decreasing time and costs associated with dioxin detection. Dioxins readily accumulate in fish tissue and fish are a common food source for humans. Thus, the goal of this research was to develop a screening technique for dioxins in fish samples using enzyme-linked immunosorbent assay (ELISA). Three approaches, each with a different fish sample purification method but all using ELISA detection, were undertaken. This research concluded that the approach of Florisil cleanup followed by ELISA detection (Florisil-ELISA) was suitable as a screening technique. The other two approaches, one using gel permeation chromatography (GPC-ELISA) and the other using acid silica and carbon columns (acid silica/carbon-ELISA) for fish sample cleanup, were not suitable.

scholarly journals Enzyme-linked immunosorbent assay (ELISA) for the screening of dioxins in fish samples

2021 ◽  
Author(s):  
Elaine Yu-Lan Chen
Keyword(s):  
Immunosorbent Assay ◽  
High Resolution Mass Spectrometry ◽  
Gel Permeation Chromatography ◽  
The Other ◽  
Alternative Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fish Sample ◽  
Purification Method ◽  
Screening Technique ◽  
Fish Samples

Dioxins are environmental contaminants that are toxic to humans. The conventional analytical method for dioxins, gas chromatography - high resolution mass spectrometry, is extremely time-consuming and expensive. Research is needed to find alternative methods that will increase sample throughput while decreasing time and costs associated with dioxin detection. Dioxins readily accumulate in fish tissue and fish are a common food source for humans. Thus, the goal of this research was to develop a screening technique for dioxins in fish samples using enzyme-linked immunosorbent assay (ELISA). Three approaches, each with a different fish sample purification method but all using ELISA detection, were undertaken. This research concluded that the approach of Florisil cleanup followed by ELISA detection (Florisil-ELISA) was suitable as a screening technique. The other two approaches, one using gel permeation chromatography (GPC-ELISA) and the other using acid silica and carbon columns (acid silica/carbon-ELISA) for fish sample cleanup, were not suitable.

scholarly journals Principles and techniques towards successful development of enzyme-linked immunosorbent assay (ELISA) for dioxin analysis

2021 ◽  
Author(s):  
Wei Zhang
Keyword(s):  
Immunosorbent Assay ◽  
Environmental Protection Agency ◽  
High Resolution Mass Spectrometry ◽  
Enzyme Linked Immunosorbent Assay ◽  
Environmental Matrices ◽  
Successful Development ◽  
Trace Level ◽  
The Us ◽  
Us Epa ◽  
Technical Details

Dioxins are highly toxic, persistent and bio-accumulative compounds. Laboratory detection of dioxins in various environmental matrices is one of the most technically demanding and expensive tasks in analytical chemistry.The cost to analyze a soil sample by conventional gas chromatography-high resolution mass spectrometry (GC-HRMS) is approximately $1,900 USC accordign to the Unisted States Environmental Protection Agency (US EPA) (Billets, 2005). As an alternative, enzyme-linked immunosorbent assay (ELISA) for dioxin analysis has been commercially available for over a decade and recognized as the US EPA Method 3025. However, assay attributes need to be examined, especially at trace level detection. In this study, sources of error in ELISA, such as background contamination and dioxin-like polycholrinated biphenyl (dl-BCB) cross-reactions have been investigated. Quality assurance data on spikes have been reviewed and the recovery was estimated to be 70%. Technical details that are crucial for the performance of dioxin in ELISA were also identified and addressed.

Indirect Enzyme-Linked Immunosorbent Assay for the Identification of Sole (Solea solea), European Plaice (Pleuronectes platessa), Flounder (Platichthys flesus), and Greenland Halibut (Reinhardtius hippoglossoides)

1999 ◽  
Vol 62 (10) ◽  
pp. 1178-1182 ◽  
Author(s):  
ANA CÉSPEDES ◽  
TERESA GARCÍA ◽  
ESTHER CARRERA ◽  
ISABEL GONZÁLEZ ◽  
ALICIA FERNÁNDEZ ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Greenland Halibut ◽  
Flatfish Species ◽  
Pleuronectes Platessa ◽  
Platichthys Flesus ◽  
Solea Solea ◽  
Fish Samples ◽  
Greenland Halibut Reinhardtius Hippoglossoides ◽  
Reinhardtius Hippoglossoides

Polyclonal antibodies produced against soluble muscle protein extracts from sole (Solea solea), European plaice (Pleuronectes platessa), flounder (Platichthys flesus), and Greenland halibut (Reinhardtius hippoglossoides) were used in an indirect enzyme-linked immunosorbent assay for the specific identification of fillets from these flatfish species. The assay was performed in two different formats: microtiter plates and immunostick tubes. Immunorecognition of antibodies adsorbed to their specific fish samples was made with goat antirabbit immunoglobulins conjugated to the enzyme horseradish peroxidase. Subsequent enzymatic conversion of the substrate allowed unequivocal identification of all flatfish species studied.

scholarly journals Principles and techniques towards successful development of enzyme-linked immunosorbent assay (ELISA) for dioxin analysis

2021 ◽  
Author(s):  
Wei Zhang
Keyword(s):  
Immunosorbent Assay ◽  
Environmental Protection Agency ◽  
High Resolution Mass Spectrometry ◽  
Enzyme Linked Immunosorbent Assay ◽  
Environmental Matrices ◽  
Successful Development ◽  
Trace Level ◽  
The Us ◽  
Us Epa ◽  
Technical Details

Dioxins are highly toxic, persistent and bio-accumulative compounds. Laboratory detection of dioxins in various environmental matrices is one of the most technically demanding and expensive tasks in analytical chemistry.The cost to analyze a soil sample by conventional gas chromatography-high resolution mass spectrometry (GC-HRMS) is approximately $1,900 USC accordign to the Unisted States Environmental Protection Agency (US EPA) (Billets, 2005). As an alternative, enzyme-linked immunosorbent assay (ELISA) for dioxin analysis has been commercially available for over a decade and recognized as the US EPA Method 3025. However, assay attributes need to be examined, especially at trace level detection. In this study, sources of error in ELISA, such as background contamination and dioxin-like polycholrinated biphenyl (dl-BCB) cross-reactions have been investigated. Quality assurance data on spikes have been reviewed and the recovery was estimated to be 70%. Technical details that are crucial for the performance of dioxin in ELISA were also identified and addressed.

Detection of Staphylococcal Enterotoxin H by an Enzyme-Linked Immunosorbent Assay

1996 ◽  
Vol 59 (3) ◽  
pp. 327-330 ◽  
Author(s):  
YI-CHENG SU ◽  
AMY C. LEE WONG
Keyword(s):  
Hydrogen Peroxide ◽  
Immunosorbent Assay ◽  
Sulfonic Acid ◽  
Culture Supernatant ◽  
Staphylococcal Enterotoxin ◽  
The Other ◽  
Enzyme Linked Immunosorbent Assay ◽  
Standard Curve ◽  
Negative Results ◽  
Enzyme Substrate

A double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the detection of a newly identified staphylococcal enterotoxin H (SEH). Peroxidase was conjugated to antibodies specific to the enterotoxin. 2,2′ Azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid)(ABTS) in hydrogen peroxide solution was used as the enzyme substrate. A standard curve of purified SEH was prepared with concentrations ranging from 1.3 to 50 ng/ml. SEH at levels equal to 2.5 ng/ml and higher were detected by this procedure. Culture supernatant from the growth of selected Staphylococcus aureus strains was analyzed by using the ELISA. SEH was produced by three of 20 strains that produced one identified enterotoxin. Ten of 21 strains, previously shown to produce substances that induced emesis in monkeys but not any known enterotoxins (A through E), were also positive for SEH production. The other 11 strains gave negative results in the ELISA, indicating that other unidentified serological types of enterotoxin exist.

scholarly journals Detection of Fertility Levels of Female Bawean Deer (Axis kuhlii) Based on Fecal Steroid Metabolic

2021 ◽  
Vol 4 (1) ◽  
pp. 84
Author(s):  
Mitha Ardila Rahmawati ◽  
Mas’ud Hariadi ◽  
Tjuk Imam Restiadi ◽  
Rimayanti Rimayanti ◽  
Tita Damayanti Lestari ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Luteal Phase ◽  
Follicular Phase ◽  
The Other ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fertility Level ◽  
Fecal Steroid ◽  
Pregnancy Status

The aim of the study was to detect the fertility level of females Bawean deer (Axis kuhlii) through fecal steroid metabolic which were tested using Enzyme-Linked Immunosorbent Assay (ELISA). The study used five females Bawean deer in Taman Flora Surabaya, which were marked using R1, R2, R3, R4, and R5 necklaces. The feces of each deer was collected on the 1st day, 6th day, 11th day, 16th day, and 21st day. The samples which were collected then extracted using the freeze dry method. Samples were tested using ELISA. The results of the measurement of the levels of fecal steroid metabolic  in five Bawean deers showed that three Bawean deer (R1, R4, and R5) were in the luteal or pregnant phase as indicated by the increased of progesterone from 1st day to 21st day. While the other two Bawean deer (R2 and R3) were in the follicular or estrous phase as indicated by the decreased of level of faecal steroid metabolic  on 11th day for deer R2 and 16th day for deer R3. The results showed that the levels of the fecal steroid metabolic can be used to determine the follicular phase and luteal phase and the pregnancy status of female Bawean deer.

scholarly journals Measurement of rotavirus antibody by an enzyme-linked immunosorbent assay blocking assay

1978 ◽  
Vol 8 (3) ◽  
pp. 283-287
Author(s):  
R H Yolken ◽  
R G Wyatt ◽  
B A Barbour ◽  
H W Kim ◽  
A Z Kapikian ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Animal Species ◽  
Assay System ◽  
The Other ◽  
New Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fixed Amount ◽  
Blocking Assay

A new method for the measurement of rotavirus antibody is described, utilizing the system of enzyme-linked immunosorbent assay (ELISA). In this method, serum is incubated with a fixed amount of rotavirus antigen, and the amount of antibody is determined by measuring the amount of unneutralized antigen. Such an assay system proved to be as efficient as the other available rotaviral antibody systems. The ELISA blocking assay also has the advantages of not requiring purified or gnotobiotic antigen and of being able to measure rotaviral antibody in all animal species.

scholarly journals Detection by PCR-Enzyme-Linked Immunosorbent Assay of Clostridium botulinum in Fish and Environmental Samples from a Coastal Area in Northern France

2002 ◽  
Vol 68 (12) ◽  
pp. 5870-5876 ◽  
Author(s):  
Patrick Fach ◽  
Sylvie Perelle ◽  
Françoise Dilasser ◽  
Joël Grout ◽  
Claire Dargaignaratz ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Environmental Samples ◽  
Clostridium Botulinum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mouse Bioassay ◽  
Type B ◽  
Probable Number ◽  
Northern France ◽  
Fish Samples ◽  
High Prevalence

ABSTRACT The prevalence of Clostridium botulinum types A, B, E, and F was determined in 214 fresh fish and environmental samples collected in Northern France. A newly developed PCR-enzyme-linked immunosorbent assay (ELISA) used in this survey detected more than 80% of samples inoculated with fewer than 10 C. botulinum spores per 25 g and 100% of samples inoculated with more than 30 C. botulinum spores per 25 g. The percent agreement between PCR-ELISA and mouse bioassay was 88.9%, and PCR-ELISA detected more positive samples than the mouse bioassay did. The prevalence of C. botulinum in seawater fish and sediment was 16.6 and 4%, respectively, corresponding to 3.5 to 7 and 1 to 2 C. botulinum most-probable-number counts, respectively, and is in the low range of C. botulinum contamination reported elsewhere. The toxin type identification of the 31 naturally contaminated samples was 71% type B, 22.5% type A, and 9.6% type E. Type F was not detected. The high prevalence of C. botulinum type B in fish samples is relatively unusual compared with the high prevalence of C. botulinum type E reported in many worldwide and northern European surveys. However, fish processing and fish preparation in France have not been identified as a significant hazard for human type B botulism.

Comparison of the Mouse Bioassay and Enzyme-Linked Immunosorbent Assay Procedures for the Detection of Type A Botulinal Toxin in Food

2004 ◽  
Vol 67 (1) ◽  
pp. 203-206 ◽  
Author(s):  
J. L. FERREIRA ◽  
S. J. ELIASBERG ◽  
P. EDMONDS ◽  
M. A. HARRISON
Keyword(s):  
Immunosorbent Assay ◽  
Lethal Dose ◽  
Type A ◽  
Foodborne Illness ◽  
The Other ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mouse Bioassay ◽  
Elisa Method ◽  
Standard Type ◽  
Standard Curve

Samples of chili linked to a foodborne illness outbreak of type A botulism were examined for preformed type A botulinal toxin using two enzyme-linked immunosorbent assay (ELISA) procedures and the mouse bioassay. One of the samples was positive for type A botulinal toxin and three of the samples were negative for type A, B, E, and F botulinal toxins using the three methods. The mouse bioassay indicated that type A toxin was present at the 10,000 minimal lethal dose per gram (MLD per g) of product. The ELISA tests indicated a toxicity of 7,650 MLD per g with one method and 8,350 MLD per g with the other method. The sample toxicity determined by the ELISA was estimated by comparing samples to a standard curve generated with standard type A neurotoxin in casein buffer. The ELISA methods are more rapid than the mouse bioassay, since the toxin type can be determined in 1 day. The mouse bioassay is more sensitive than the ELISA but usually requires multiple assays to obtain the toxin type and toxicity. Type A culture isolates from the sample were also verified using one ELISA method.

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