scholarly journals MEASUREMENT OF SERUM TESTOSTERONE IN KACANG GOAT BYUSING ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) TECHNIQUE: THE IMPORTANCE OF KIT VALIDATION

2016 ◽  
Vol 10 (1) ◽  
Author(s):  
Gholib G ◽  
Sri Wahyuni ◽  
Okta Hilda Kadar ◽  
Mulyadi Adam ◽  
Triva Murtina Lubis ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Jugular Vein ◽  
Serum Testosterone ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Validation ◽  
Validation Data ◽  
Testosterone Concentration ◽  
Standard Curve ◽  
Biological Validation ◽  
Elisa Technique

This study was conducted to validate a commercial testosterone enzyme-linked immunosorbent assay (ELISA) kits (DRG EIA-1559) inanalytic and biological manner for measuring serum testosterone concentrations in kacang goats. This study used 18 healthy kacang goats, sixbucks (>2 years), six kids (<6 months), and six does (>2 years). Blood samples were collected from jugular vein and prepared as serum. Twovalidation tests were performed, an analytical validation comprises a parallelism, accuracy, precision and sensitivity and a biological validationby comparing testosterone concentration from bucks, kids, and does. Testosterone concentrations were measured using ELISA technique. Data ofanalytical validation were analyzed descriptively and test of equality of slope was performed to see the parallelism between samples and standardcurves. Analysis of variance (ANOVA) was used for biological validation data. Results of parallelism showed that sample curve was parallel tothe standard curve. Accuracy, precision (% CV of intra-and inter-assay) and sensitivity of the assay were 99.65±4.27%, <10%, <15% and 0.083ng/ml, respectively. Results of biological validation showed that the assay used were accurately measured testosterone which testosteroneconcentrations in bucks were significantly higher compared to kids and does (P<0.05). In conclusion, a commercial testosterone ELISA kits(DRG EIA-1559) is a reliable assay for measuring serum testosterone concentration in kacang goats. Key words: analytical and biological validations, ELISA, testosterone, kacang goat

Enzyme-Linked Immunosorbent Assay for Microcystins in Blue-Green Algal Blooms

1990 ◽  
Vol 73 (3) ◽  
pp. 451-456 ◽  
Author(s):  
Fun S Chu ◽  
Xuan Huang ◽  
R D Wei
Keyword(s):  
Liquid Chromatography ◽  
Immunosorbent Assay ◽  
Algal Blooms ◽  
Ammonium Bicarbonate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Standard Curve ◽  
Green Algal ◽  
Recovery Study ◽  
Minimum Detection Level ◽  
Elisa Plate

Abstract A direct competitive enzyme-linked immunosorbent assay (ELISA) for the freshwater blue-green algal toxin mlcrocystln (MCYST) In algae and water was developed. The assay Involves coating antl-MCYST-variant leuclne-arglnine (LR) antibody to the ELISA plate and the use of MCYST-LRperoxidase as the enzyme marker. The linear portion of the standard curve for MCYST in phosphate buffer containing saline (PBS) was 0.5-10.0 ng/mL (25-500 pg/assay). The minimum detection level for MCYST-LR was 0.20 ng/mL (10 pg/assay). Contaminated water could be directly used In the ELISA. The overall analytical recoveries for MCYST-LR added to water at levels of 1-20 ng/mL was 83.4%. For analysis of cellular MCYST, the toxin was first extracted from the algae with 0.1M ammonium bicarbonate, diluted with PBS to less than 0.5 mg dried algae/mL (&lt;5.0 mg wet welght/mL) and directly used in the ELISA. C-18 reverse-phase Sep-Pak cartridges effectively adsorbed MCYST from the toxln-containlng solutions. The toxin could be recovered from the cartridge by elutlng with 60% methanol. Using this approach, an algae extract that was relatively free of MCYST was prepared and was used in a recovery study. The overall analytical recovery of MCYST added to the algae extract In the range of 0.25-20 ppm was 83% with a coefficient of variation of 11.9%. The detection limit for MCYST In dried algae was about 0.25-0.5 pg/g (0.25-0.5 ppm) lyophlllzed algae sample. This method was applied for the analysis of several naturally occurring algal blooms. Limited samples were also analyzed for MYCST by liquid chromatography. ELISA data were in general agreement with those obtainedby liquid chromatography. MCYST concentrations from 0.006 to 2.9 fig/g (6 to 2900 ppb) and from 26 to 5200 /ig/g (26 ppm to 5200 ppm) were found In water and algae (dried weight), respectively

Analytical validation of an enzyme‐linked immunosorbent assay for the quantification of S100A12 in the serum and feces of cats

2019 ◽  
Vol 48 (4) ◽  
pp. 754-761
Author(s):  
Cory S. Bridges ◽  
Niels Grützner ◽  
Jan S. Suchodolski ◽  
Jörg M. Steiner ◽  
Romy M. Heilmann
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Validation

scholarly journals Aflatoxin Plate Kit

2011 ◽  
Vol 94 (5) ◽  
pp. 1519-1530
Author(s):  
Arthur Trombley ◽  
Titan Fan ◽  
Robert LaBudde
Keyword(s):  
Immunosorbent Assay ◽  
Hplc Method ◽  
Aflatoxin Contamination ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rapid Testing ◽  
Total Aflatoxin ◽  
Standard Curve ◽  
Independent Laboratory ◽  
Test Kit

Abstract The level of total aflatoxin contamination was analyzed in naturally contaminated and spiked samples of corn and peanut using the Aflatoxin Plate Kit. This kit is an enzyme-linked immunosorbent assay (ELISA) suitable for rapid testing of grains and peanuts. The assay was evaluated for ruggedness and linearity of the standard curve. The test kit results were then statistically evaluated for accuracy, precision, and correlation to a validated HPLC method (AOAC 994.08). The results were verified by an independent laboratory.

scholarly journals Single-Dilution Enzyme-Linked Immunosorbent Assay for Quantification of Antigen-Specific Salmonid Antibody

2000 ◽  
Vol 12 (3) ◽  
pp. 245-252 ◽  
Author(s):  
Stewart W. Alcorn ◽  
Ronald J. Pascho
Keyword(s):  
Immunosorbent Assay ◽  
Sockeye Salmon ◽  
High Titer ◽  
Repeated Measurements ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Activity ◽  
Standard Curve ◽  
Test Fish ◽  
The Mean ◽  
Antibody Levels

An enzyme-linked immunosorbent assay (ELISA) was developed on the basis of testing a single dilution of serum to quantify the level of antibody to the p57 protein of Renibacterium salmoninarum in sockeye salmon ( Oncorhynchus nerka). The levels of antibody were interpolated from a standard curve constructed by relating the optical densities (OD) produced by several dilutions of a high-titer rainbow trout ( O. mykiss) antiserum to the p57 protein. The ELISA OD values produced by as many as 36 test sera on each microplate were compared with the standard curve to calculate the antigen-specific antibody activity. Repeated measurements of 36 samples on 3 microplates on each of 6 assay dates indicated that the mean intraassay coefficient of variation (CV) was 6.68% (range, 0–23%) and the mean interassay CV was 8.29% (range, 4–16%). The antibody levels determined for the serum sample from 24 sockeye salmon vaccinated with a recombinant p57 protein generally were correlated with the levels determined by endpoint titration ( r2 = 0.936) and with results from another ELISA that was based on extrapolation of antibody levels from a standard curve ( r2 = 0.956). The single-dilution antibody ELISA described here increases the number of samples that can be tested on each microplate compared with immunoassays based on analysis of several dilutions of each test serum. It includes controls for interassay standardization and can be used to test fish weighing <3 g.

scholarly journals Enzyme Linked Immunosorbent Assay for Fumonisin B1 Detection in Local Corn Seeds from Baghdad-Iraq

2021 ◽  
pp. 4621-4627
Author(s):  
Sinai W. Mohammed ◽  
Hanan J. Nayyef ◽  
Fadhaa O. Sameer ◽  
Ahmed Y. Hanoon
Keyword(s):  
Relative Density ◽  
Immunosorbent Assay ◽  
Fumonisin B1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fusarium Spp ◽  
Isolation And Identification ◽  
Toxic Compounds ◽  
Identification Of Fungi ◽  
Elisa Technique ◽  
Cancer Activity

Fungi produce a series of toxic compounds on corn, especially Fumonisin B1 (FB1) toxin produced by Fusarium spp. and promoting cancer activity in humans and animals. This study aimed to the isolation and identification of fungi associated with local corn seeds and the detection for the presence of FB1 by using ELISA technique. Thirty samples of corn ears were collected from silos and markets in Baghdad city during the period from November 2018 to March 2019. The present study found that Fusarium was the dominant isolate among fungi in terms of the relative density 57.07%, followed by Aspergillus 31.17%, Rhizopus 3.36%, Alternaria 2.88%, Mucor 2.16%, Penicillium 1.92%, Trichothecium 0.96%, and Helminthosporium 0.48%. FB1 was detected in all samples of the silos and markets with a concentration range of 13.69 - 175.54 µg/kg. There were no significant differences in FB1concentration among samples collected from the silos and markets. Also, no relationship was found between the number of infected seeds by Fusarium spp. and FB1concentrations.

scholarly journals Enzyme Linked Immunosorbent Assay (ELISA) Technique Guideline

2021 ◽  
Vol 5 (5) ◽  
pp. 447-453
Author(s):  
Rachmat Hidayat ◽  
Patricia Wulandari
Keyword(s):  
Immunosorbent Assay ◽  
Solid Surface ◽  
Color Change ◽  
Antibody Binding ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hormone Levels ◽  
Elisa Technique ◽  
Antibody Complex ◽  
Antigen Antibody ◽  
Detection Signal

ELISA (Enzyme-linked immunosorbent assay) is a technique used to assess the quantification of peptide, protein, antibody and hormone levels, based on the principle of antigen-antibody binding. In the ELISA technique, antigen immobilization will be carried out on a solid surface, then bound with antibodies to form an antigen-antibody bond complex, where the antigen-antibody complex is bound to the enzyme. The detection signal in the form of a color change will be formed due to the reaction between the enzyme and the substrate.

Sign in / Sign up

Export Citation Format

Share Document