Development of Direct Competitive ELISA Kit for the Detection of Tetrodotoxin Using HRP Labeled Antigen

2011 ◽  
Vol 236-238 ◽  
pp. 2820-2824 ◽  
Author(s):  
Qing Ping Zhong ◽  
An Cheng Huang ◽  
Bin Wang ◽  
Xue Dong
Keyword(s):  
Room Temperature ◽  
Inhibitory Concentration ◽  
Enzyme Linked Immunosorbent Assay ◽  
Competitive Reaction ◽  
Antigen Concentration ◽  
Time Saving ◽  
Elisa Kit ◽  
Standard Curve ◽  
Coating Method ◽  
Ml Detection

The direct competitive enzyme-linked immunosorbent assay (dcELISA) kit was developed for detecting tetrodotoxin (TTX). The working conditions of the dcELISA kit including the anti-TTX mAb coating concentration, coating method, enzyme-labeled antigen concentration, the antigen diluents, reaction time and temperature were all optimized. The result showed that mAb coating concentration was 3.72 μg/ml, it was coated at the condition of minimal power treatment of microwave oven for 3 min. The enzyme-labeled antigen concentration was 4.08 μg/ml. The competitive reaction was under the condition of room temperature 25 °C for 30 min. The half maximal inhibitory concentration (IC50) of the standard curve was 20.4 ng/ml, detection limit was 1.1 ng/ml, linear range 3.3～137 ng/ml, the intra-assay CV and inter-assay CV were 6.25% and 7.34% respectively. And recovery rate of TTX ranged from 65.0% to 93.2% with the CV of 9.41～12.77%. This method is convenient, sensitive and time-saving, hope this dcELISA kit can bring benefits and reference for TTX detection.

scholarly journals Validation of Commercial ELISA kit for Non-Invasive Measurement of Cortisol Concentrations and the Evaluation of the Sampling Time of Blood and Fecal Sample in Aceh Cattle

2020 ◽  
Vol 151 ◽  
pp. 01007
Author(s):  
Gholib Gholib ◽  
Sri Wahyuni ◽  
Ahmad Wahyudi ◽  
Khoiriyah S. Silalahi ◽  
Muslim Akmal ◽  
...  
Keyword(s):  
Diurnal Variation ◽  
Fecal Sample ◽  
Plasma Cortisol ◽  
Sampling Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sample Collection ◽  
Elisa Kit ◽  
Standard Curve ◽  
Fecal Cortisol ◽  
Significant Difference

Some parameters should be evaluated before the analysis of cortisol hormone using enzyme-linked immunosorbent assay ( ELISA ). The most important one is to test the validity of the ELISA kit itself since most of the commercially available ELISA kit is not designed for animal use. The time of sample collection is also another important parameter needed to be evaluated due to a known diurnal variation of cortisol concentrations. This study aims to validate a commercial ELISA kit and evaluate the sampling time on the diurnal variations of cortisol concentration in blood and feces of Aceh cattle. Blood and fecal samples were collected in the morning and afternoon from 8 Aceh cows aged 2-3 years. A commercial ELISA Kit (Cat. No. EIA-K003-H5, Arbor Assays®) was validated using analytical (parallelism, accuracy, and precision/% CV of intraand inter-assay) and biological validations (by analyzing sample preand post-transportation). The results of a parallelism test showed the diluted sample curve from the fecal extract was not significantly different (parallel) to the standard curve of EIA-K003-H5 kits (P>0.05), the accuracy of the assay: 99,76 ± 3.77%, and the % CV of intra-and inter-assay less than 10%. The results from the biological validation test showed that the concentrations of fecal cortisol post-transportation were significantly higher compared to the pre-transportation (P<0.05). Plasma cortisol concentrations collected in the morning were significantly higher (79.34%) compared to afternoon samples(P<0.05). However, cortisol concentrations in feces collected in the morning and afternoon did not show a significant difference (P>0.05). In conclusion, ELISA Kit (EIA-K003H5, Arbor Assays®) is a reliable assay for measuring cortisol in the feces of Aceh cows. Plasma cortisol concentrations in Aceh cows show a diurnal variation which is higher in the morning than afternoon, but the concentration of cortisol in the fecal sample did not.

Can Osteopontin Be Considered a Biomarker for Endometriosis?

2017 ◽  
Vol 68 (9) ◽  
pp. 2132-2134
Author(s):  
Daniela Roxana Albu (Matasariu) ◽  
Elena Mihalceanu ◽  
Alina Pangal ◽  
Carmen Vulpoi ◽  
Mircea Onofriescu ◽  
...  
Keyword(s):  
Serum Level ◽  
Immunosorbent Assay ◽  
Pelvic Pain ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Multifactorial Disease ◽  
Elisa Kit ◽  
Healthy Women ◽  
Progesterone Treatment ◽  
Large Dispersion

Endometriosis is a multifactorial disease that is manifested by infertility and pelvic pain. The purpose of the study was to evaluate the effect of progesterone treatment on the serum level of osteopontin, a multipotent cytokine, in patients with endometriosis. The study was prospective and we evaluated osteopontin levels that were measured in the serum of 40 patients with endometriosis and 12 healthy women using a standardized Enzyme-Linked Immunosorbent Assay (ELISA) kit. Osteopontin seric levels were lower in endometriosis patients and increased after progesterone treatment. Because of the large dispersion of data even in the control group, we find the association between osteopontin and endometriosis questionable.

In-situ construction of direct Z-scheme AgBr/α-Ag2WO4 heterojunction with promoted spatial charge migration and photocatalytic performance

2021 ◽  
Author(s):  
Xiao-Ya Zhai ◽  
Yifan Zhao ◽  
Guo-Ying Zhang ◽  
Bing-Yu Wang ◽  
Qi-Yun Mao
Keyword(s):  
Anion Exchange ◽  
Large Scale ◽  
Room Temperature ◽  
Photocatalytic Performance ◽  
Charge Migration ◽  
Time Saving ◽  
Spatial Charge ◽  
Construction Strategy ◽  
Large Scale Synthesis

In the work, a direct Z-scheme AgBr/α-Ag2WO4 heterojunction was prepared by in-situ anion exchange at room temperature. The construction strategy is energy- and time-saving for large scale synthesis. The α-Ag2WO4...

scholarly journals Evaluation of an enzyme immunoassay for clinical diagnosis of neurocysticercosis in symptomatic patients

2010 ◽  
Vol 43 (6) ◽  
pp. 647-650 ◽  
Author(s):  
Reynaldo Mendes de Carvalho Junior ◽  
Dorcas Lamounier Costa ◽  
Savyo Carvalho Soares ◽  
Carlos Henrique Nery Costa
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Sensitivity And Specificity ◽  
Cerebral Spinal Fluid ◽  
Taenia Solium ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Disease ◽  
Multivariate Logistic Regression ◽  
Human Central Nervous System ◽  
Elisa Kit

INTRODUCTION: Neurocysticercosis is an infection of the human central nervous system caused by the metacestode larvae of Taenia solium. Neurocysticercosis is the most common parasitic disease in developing countries. Epilepsy is the most common clinical manifestation. Difficulties in confirming the diagnosis motivated the evaluation of the enzyme-linked immunosorbent assay on cerebral spinal fluid (CSF). METHODS: Twenty-two patients with NCC and 44 control patients were studied. CSF was analyzed using a commercial ELISA kit developed for NCC. Sensitivity and specificity were measured and a multivariate logistic regression was performed. RESULTS: Sensitivity and specificity of ELISA were 31.8% and 100%, respectively, with accuracy of 77.3%. Only the size of the lesions proved to be important for performance of the test. CONCLUSIONS: The results showed that ELISA contributes to the diagnosis of neurocysticercosis if the result is negative or if the patient has a lesion of 2 cm or more.

scholarly journals COMPARISON OF ANTIVENOM POTENTIAL OF CHICKEN EGG YOLK ANTIBODIES GENERATED AGAINST BENTONITE AND ADJUVENT COATED VENOMES OF COMMON POISONOUS SNAKE IN INDIA

1970 ◽  
Vol 7 (1) ◽  
pp. 259-267 ◽  
Author(s):  
S. Meenatchisundaram ◽  
R. Selvakumaran ◽  
G. Parameswari ◽  
A. Michael
Keyword(s):  
Room Temperature ◽  
Deae Cellulose ◽  
Egg Yolk ◽  
Precipitation Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Snake Venoms ◽  
Chicken Egg ◽  
Ammonium Sulphate Precipitation ◽  
Egg Yolk Antibodies ◽  
Chicken Egg Yolk

Antivenom antibodies were generated in white leghorn chicken against bentonite and adjuvant coated venoms of Common Indian Poisonous Snakes (Cobra, Krait, Russell's viper and Saw Scaled viper).The antivenom from immunized chicken egg yolk were purified by polyethylene glycol (PEG) and ammonium sulphate precipitation method and further purified by DEAE cellulose ion exchange column chromatography and concentrated by polyvinyl pyrolidone powder at room temperature. The titer of antibodies was estimated using Enzyme Linked Immunosorbent Assay (ELISA).The chickens immunized with Freund's complete adjuvant showed slightly higher titre when compared to bentonite. Inhibition of lethal, edema, haemorrhagic, procoagulant and phospholipase A2 and fibrinolytic activities of snake venoms were determined. The chicken egg yolk antivenom was effective in neutralization of these toxic and enzymatic activities of venom. The median effective dose (ED50) of chicken egg yolk antibodies raised against adjuvant coated venoms showed effective neutralizing venom toxicity when compared to the antibodies raised using bentonite coated venoms.

scholarly journals Investigating the dynamics of Leishmania antigen in the urine of patients with visceral leishmaniasis: a pilot study

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 1514
Author(s):  
Prakash Ghosh ◽  
Israel Cruz ◽  
Albert Picado ◽  
Thomas Edwards ◽  
Md. Anik Ashfaq Khan ◽  
...  
Keyword(s):  
Pilot Study ◽  
Visceral Leishmaniasis ◽  
Treatment Monitoring ◽  
Urine Samples ◽  
Urinary Antigen ◽  
Antigen Concentration ◽  
Elisa Kit ◽  
Time Points ◽  
Non Invasive ◽  
Antigen Elisa

Background: Detection of Leishmania antigens in the urine provides a non-invasive means of diagnosis and treatment monitoring of cases of visceral leishmaniasis (VL). Leishmania antigen load in the urine may vary between different time-points within a day, thus influencing the performance of antigen-detection tests. Methods: We investigated the dynamics of Leishmania antigen in urine collected at three different time points (08:00, 12:00 and 16:00 hours). All urine samples collected were tested with the Leishmania Antigen ELISA (VL ELISA) kit, produced by Kalon Biological Ltd., UK. Results: The median concentration of Leishmania antigen in urine collected at 08:00 (2.7 UAU-urinary antigen units/ml) was higher than at 12:00 (1.7 UAU/ml) and at 16:00 (1.9 UAU/ml). These differences were found to be statistically significant (08:00 vs. 12:00, p=0.011; 08:00 vs. 16:00, p=0.041). Conclusion: This pilot study indicates that the Leishmania antigen concentration is higher in urine samples collected in the morning, which has important implications when the VL ELISA kit or other tests to detect Leishmania antigen in urine are used for diagnosis of VL and treatment monitoring.

scholarly journals Kadar Prostaglandin dan Endorfin pada Remaja dengan Dismenore Primer yang diberi Hidroterapi Hangat dan Dingin

2020 ◽  
Vol 12 (1) ◽  
pp. 115-121
Author(s):  
Andi Asrina ◽  
Arsyad Aryadi ◽  
Nilawati Andi
Keyword(s):  
Experimental Study ◽  
Enzyme Linked Immunosorbent Assay ◽  
Primary Dysmenorrhea ◽  
Not Given ◽  
Elisa Kit ◽  
Group Design ◽  
Prostaglandin Level ◽  
Post Test ◽  
Quasi Experimental ◽  
The Mean

This study aims to determine the comparison of prostaglandin and endorphin levels in adolescents with primary dysmenorrhea with and without warm (37-40oC) and cold (18-20oC) hydrotherapy. This quasi-experimental study with a post-test only controls group design was carried out in Islamic Boarding Schools with a sample of 36 young girls divided into 3 groups: 12 teens given warm hydrotherapy, 12 teens given cold hydrotherapy and 12 teens not given intervention (control). Blood plasma is taken after an intervention is given on the first day of menstruation. Examination of prostaglandin and endorphins levels using the enzyme-linked immunosorbent assay (ELISA) kit method. After cold hydrotherapy, the mean levels of prostaglandins in the cold hydrotherapy group were twice higher (569 pg/ml) compared to controls (394 pg/ml). The mean prostaglandin level in the warm hydrotherapy group also showed an increase prostaglandin (437 pg/ml) compared to the control (394 pg/ml). In addition to increasing levels of prostaglandins, increased levels of endorphins also occurred in the group given warm hydrotherapy (154 pg/ml) and the group was given cold hydrotherapy (187 pg/ml) compared to the control (119 pg/ml) p = 0.001. The conclusion in this study is that warm and cold hydrotherapy can increase levels of prostaglandins and endorphins in adolescents with primary dysmenorrhea. However, cold hydrotherapy increases endorphin levels higher than warm hydrotherapy. Key words: Prostaglandin, Endorphin, Hydrotherapy, Primary Dismenorrhea.

scholarly journals ANTIFUNGAL ACTIVITY OF MANGIFERA INDICA LEAF EXTRACT ON FUNGI ISOLATED FROM BREAD VENDED WITHIN BAKORI, NIGERIA

2021 ◽  
pp. 1-4
Author(s):  
ABDULAZIZ BASHIR KUTAWA ◽  
SALMAN SADA BAKORI ◽  
HABU MUSA
Keyword(s):  
Antifungal Activity ◽  
Room Temperature ◽  
Inhibitory Concentration ◽  
Mangifera Indica ◽  
Morphological Characteristics ◽  
Diffusion Method ◽  
Leaf Extracts ◽  
Chemical Test ◽  
Concentration Result ◽  
Powdered Form

Objective: The present study was carried out to determine the antifungal activity of Mangifera indica leaves extract on fungi isolated from bread vended within Bakori. Methods: The powdered form of M. indica leaves was used to prepare the extract using ethanol, the leaves were air dried at room temperature for 10 days. Results: The chemical test was carried out to identify the secondary metabolites, some of the metabolites that were present include alkaloids, saponins, flavonoids, steroids, and tannins. Both fungal isolates were identified on the basis of morphological characteristics as Aspergillus niger, Aspergillus flavus, and Mucor spp. The antifungal activity of M. indica leaves extract was determined using agar well diffusion method on Aspergillus and Mucor spp. The results showed that the extract was found to inhibit A. niger at 100 mg/ml, 50 mg/ml, and 25 mg/ml with 13.9 mm, 11.5 mm, and 8.0 mm, respectively, and A. flavus at 100 mg/ml, 50 mg/ml, and 25 mg/ml with 13.6 mm, 11.2 mm, and 8.1 mm, respectively, while Mucor spp. was found to be resistant at 25 mg/ml while 100 mg/ml and 50 mg/ml showed an activity. Minimum inhibitory concentration result showed a promising activity against Aspergillus spp. at 25 mg/ml while Mucor spp. at 50 mg/ml. Conclusion: Therefore, M. indica leaf extracts can be used in the treatment of diseases or illness caused by Aspergillus and Mucor spp.

scholarly journals Quantification of a novel dense granule protein (granulophysin) in platelets of patients with dense granule storage pool deficiency

Blood ◽  
1992 ◽  
Vol 80 (5) ◽  
pp. 1231-1237 ◽  
Author(s):  
A Shalev ◽  
G Michaud ◽  
SJ Israels ◽  
A McNicol ◽  
S Singhroy ◽  
...  
Keyword(s):  
Membrane Vesicles ◽  
Dense Granule ◽  
Enzyme Linked Immunosorbent Assay ◽  
Granule Formation ◽  
Standard Curve ◽  
Storage Pool ◽  
Granule Membrane ◽  
Hermansky Pudlak Syndrome ◽  
Novel Protein

An antigen-capture sandwich enzyme-linked immunosorbent assay (ELISA) was developed for a novel protein granulophysin, a constituent of the platelet dense granule (DG) membrane and used to characterize patients with dense granule storage pool deficiency (delta-SPD). The assay uses two monoclonal antibodies against the protein, one of which is conjugated to peroxidase. Purified DGs, an enriched source of the protein, were used for the standard curve. Granulophysin levels were only low in forms of delta-SPD associated with albinism. Granulophysin levels in platelet homogenates of 30 patients with the Hermansky-Pudlak syndrome form of delta-SPD were 1/4 to 1/5 of levels in controls or obligate heterozygotes. Two patients with the Chediak-Higashi form of delta-SPD syndrome also had markedly reduced levels of granulophysin. Patients with other forms of delta-SPD had normal levels of granulophysin. Two sisters with delta-SPD in one family had normal granulophysin present in empty dense granule membrane vesicles. Three members of another family with delta-SPD had low DG counts but normal granulophysin levels, indicating that in this group the level of granulophysin was maintained despite the reduction in granule formation. Thus, granulophysin quantitation facilitates characterization of delta-SPD patients and may provide clues to the nature of defective granules in delta-SPD subtypes.

scholarly journals Technical viability of the YF MAC-HD ELISA kit for use in yellow fever-endemic regions

2021 ◽  
Vol 15 (6) ◽  
pp. e0009417
Author(s):  
Christin H. Goodman ◽  
Maurice Demanou ◽  
Mick Mulders ◽  
Jairo Mendez-Rico ◽  
Alison Jane Basile
Keyword(s):  
South America ◽  
Yellow Fever ◽  
Accurate Diagnosis ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Kit ◽  
Wash Buffer ◽  
Antibody Capture ◽  
Vaccination Campaigns ◽  
Arboviral Disease

Yellow fever (YF), an arboviral disease, affects an estimated 200,000 people and causes 30,000 deaths per year and recently has caused major epidemics in Africa and South America. Timely and accurate diagnosis of YF is critical for managing outbreaks and implementing vaccination campaigns. A YF immunoglobulin M (IgM) antibody-capture (MAC) enzyme-linked immunosorbent assay (ELISA) kit, the YF MAC-HD, was successfully introduced starting in 2018 to laboratories in Africa and South America. The YF MAC-HD kit can be performed in 3.5 hours, test up to 24 samples, and includes all reagents necessary to perform the test, except for water used to dilute wash buffer. In 2018 and 2019, a total of 56 laboratory personnel from 39 countries in Africa and South America were trained to use the kit during workshops, followed by take-home YF IgM proficiency testing (PT) exercises. Participants received either a 10- or 20-sample YF PT panel and performed testing using the YF MAC-HD kit. All countries obtained 90% or higher correct results. These results verified the technical viability and transferability of YF MAC-HD kit use for laboratories in YF-endemic countries.

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