Testing Peanut Seeds for Peanut Stripe Virus1

1986 ◽  
Vol 13 (1) ◽  
pp. 38-40 ◽  
Author(s):  
J. W. Demski ◽  
D. Warwick
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Seed ◽  
Peanut Stripe Virus ◽  
Elisa Technique

Abstract Peanut seeds were tested by the enzyme-linked immunosorbent assay (ELISA) for peanut stripe virus without affecting their viability. A portion of cotyledon (0.02–0.05 g) was removed from the end of the seed opposite the radicle and triturated in an antigen buffer. Virus was frequently detected in extracts of cotyledons and embryos but seldom in the testa. Portions of one infected seed in ten could be detected. Correlations among ELISA, infectivity assays, and growing out tests were obtained. The ELISA technique has the advantage of detecting individual infected seed allowing the elimination of the infected ones. Identified infected seed can be used for preservation of virus cultures.

A Comparison of an Enzyme Linked Immunosorbent Assay (ELISA) and Western Blotting for Detection of Peanut Mottle Virus and Peanut Stripe Virus1

1986 ◽  
Vol 13 (2) ◽  
pp. 64-67 ◽  
Author(s):  
J. L. Sherwood ◽  
H. A. Melouk
Keyword(s):  
Western Blotting ◽  
Immunosorbent Assay ◽  
Protein A ◽  
Mottle Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coat Proteins ◽  
Western Blots ◽  
Peanut Stripe Virus ◽  
Dry Milk ◽  
The Difference

Abstract Western blotting was used to detect infections of peanut cv. Tamnut 74 with peanut mottle virus (PMV) and/or peanut stripe virus (PStV). Leaf samples were ground in electrophoresis sample buffer and heated for 5 min at 95 C prior to electrophoresis in 12% polyacrylamide gels. After electrophoresis, proteins were transferred to nitrocellulose sheets at 100V for 45 min. Western blots were performed by first blocking unbound sites on the nitrocellulose with 5% non-fat dry milk in Tris-buffered saline (TBS), pH 7.4 for 30 min, followed by incubation in a 1/200 dilution of PMV and/or PStV antiserum in TBS (the latter antiserum provided by J. W. Demski, U. of GA) for 45 min. This was followed by incubation in protein-A-peroxidase (2 μg/mL in TBS) for 45 min, followed by 4-chloro-1-napthol plus hydrogen peroxide in TBS. As little as 25 ng of either purified PMV or PStV was detected. This was similar to the limits of detection fo the double sandwich enzyme linked immunosorbent assay (ELISA). Because of the difference in migration of the coat proteins of PMV and PStV, both viruses may be detected in plants infected with PMV and PStV. This assay can be performed in approximately 6 h when mini-gels are used for the initial electrophoretic seperation and does not require the antiserum to be fractionated or bound to an enzyme as is the case with ELISA.

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.

scholarly journals Comparison of Estrous Performance and Progesterone Level of Kacang Goats Induced by PGF2α Versus Ovsynch Protocol

2020 ◽  
Vol 151 ◽  
pp. 01045
Author(s):  
Budianto Panjaitan ◽  
Almira Dewi ◽  
Fadli FR Nasution ◽  
Mulyadi Adam ◽  
Tongku N. Siregar ◽  
...  
Keyword(s):  
Estrous Cycle ◽  
Immunosorbent Assay ◽  
Group Versus ◽  
Visual Observation ◽  
Progesterone Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Progesterone Concentration ◽  
Blood Samples ◽  
Elisa Technique ◽  
K2 Group

This study aimed to compare estrous performance and progesterone concentration during the estrous cycle of kacang goats induced by PGF2α versus ovsynch protocol. This research used six female kacang goats. The goats were divided into two groups namely the K1 group which was induced by injecting 7.5 mg PGF2α intramuscularly in 10 days interval and K2 group which was induced by using the ovsynch method. Protocol for K2 group was as follow; at day 1, the goats were injected with 7.5 mg PGF2α; at day 8, they were injected with 50 µg GnRH; at day 15 they were injected with 7.5 mg PGF2α; at day 18 they were injected with 50 µg GnRH. Estrous were detected using male goat and visual observation. The blood samples were taken on days 7, 14, and 21 after estrous. Progesterone concentration was measured with enzyme-linked immunosorbent assay (ELISA) technique. Intensity, onset, and duration of estrous in K1 group versus K2 group were 8.33±2.08 vs 7.00±1.00; 56.00±34.12 vs 36.00±20.78 hours; and 24.00±26.15 vs 24.00±20.78 hours, respectively (p>0.05). Level of progesterone hormone on day 7, 14, and 21 for K1 vs K2 were 0.812±0.710 vs 2.369±3.351; 5.051±7.754 vs 3.091±4.385 ng/ml; and 4.173±6.692 vs 3.562±4.113 ng/mL, respectively (p>0,05). It can be concluded that the differences in synchronization protocols between PGF2α versus ovsynch do not affect the performance of estrous and the concentration of progesterone during the estrous cycle of kacang goats.

The enzyme-linked immunosorbent assay (ELISA) test for the identification of blood-meals of haematophagous insects

1986 ◽  
Vol 76 (2) ◽  
pp. 321-330 ◽  
Author(s):  
M. W. Service ◽  
A. Voller ◽  
D. E. Bidwell
Keyword(s):  
Field Test ◽  
Immunosorbent Assay ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fresh Blood ◽  
Elisa Technique ◽  
Blood Meals

AbstractA sandwich enzyme-linked immunosorbent assay (ELISA) test was developed to detect blood-meals in insects and identify the host fed on. The test proved both sensitive and specific. Very small quantities of fresh blood (about 0·02 μl) can be detected; in practice, this enables blood in mosquitoes which are about three-quarters gravid to be identified. In trials in both Zambia and Britain, positive reactions were easily identified visually; consequently, this enabled the ELISA technique to be used as a routine field test. In addition to those of mosquitoes, blood-meals of a few Culicoides species were also successfully identified.

Structural Aspects of Protein Accumulation in Developing Pea Cotyledons. III. Immunocytochemical Localization of Legumin and Vicilin Using Antibodies Shown to Be Specific by the Enzyme-Linked Immunosorbent Assay (ELISA)

1980 ◽  
Vol 7 (3) ◽  
pp. 339 ◽  
Author(s):  
S Craig ◽  
A Millerd ◽  
DJ Goodchild
Keyword(s):  
Immunosorbent Assay ◽  
Storage Proteins ◽  
Protein Body ◽  
Protein Bodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Accumulation ◽  
Immunocytochemical Localization ◽  
Elisa Technique ◽  
Immunocytochemical Techniques ◽  
Structural Aspects

The site of sequestration of the storage proteins legumin and vicilin during development of cotyledons from pea (Pisum sativum L.) has been determined using improved immunocytochemical techniques. Antibodies to legumin and vicilin were made monospecific by affinity chromatography. They were allowed to react on sections of glycol methacrylate-embedded cotyledon tissue and detected by indirect immunocytochemical localization using rhodamine-labelled antibodies. The enzyme-linked immunosorbent assay (ELISA) technique was adapted to verify antibody specificity at a sensitivity up to 300 times greater than that of immunodiffusion. Legumin and vicilin 4 are localized in small peripheral deposits within large vacuoles as early as day 8 after flowering. As the vacuoles fragment during development the storage proteins continue to be localized in the vacuolar deposits until, at day 16, they entirely fill vacuoles, now termed protein bodies. Thereafter, the protein bodies become more densely packed and retain a similar form from day 22 to maturity. Wherever the same vacuolar deposit of protein body could be observed in adjacent sections, antilegumin and antivicilin 4 labelled both deposits, clearly indicating that both storage proteins are sequestered into the same area of protein.

Enzyme immunoassay for factor VIII-related antigen.

1982 ◽  
Vol 28 (6) ◽  
pp. 1356-1358 ◽  
Author(s):  
J Cejka
Keyword(s):  
Regression Analysis ◽  
Factor Viii ◽  
Enzyme Immunoassay ◽  
Immunosorbent Assay ◽  
Present Method ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasma Samples ◽  
Factor Viii Related Antigen ◽  
Elisa Technique

Abstract I describe a simple enzyme-linked immunosorbent assay (ELISA) for the quantitation of Factor VIII-related antigen in plasma with use of commercially available peroxidase-labeled antiserum and solid-phase support. Regression analysis of 85 plasma samples analyzed by this technique (y) and by a commonly used electroimmunoassay (Anal. Biochem. 15: 45-52, 1966) (x) gave the equation y = 0.223 + 0.77x (r = 0.973). The present method was also compared with enzyme immunoassay in which a phosphatase-labeled antiserum prepared in our laboratory was used; the correlation between the two assays was very good. The simplicity and specificity of the ELISA technique should make it a useful alternative to the more difficult and time-consuming Laurell method.

Concentrations of apoprotein CII, CIII, and E in total serum and in the apoprotein B-containing lipoproteins, determined by a new enzyme-linked immunosorbent assay

1990 ◽  
Vol 36 (12) ◽  
pp. 2047-2052 ◽  
Author(s):  
N Alsayed ◽  
R Rebourcet ◽  
J Chapman
Keyword(s):  
Immunosorbent Assay ◽  
Healthy Subjects ◽  
Direct Determination ◽  
Total Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Particle Structure ◽  
Apo B ◽  
Apoprotein B ◽  
Elisa Technique

Abstract We describe a new enzyme-linked immunosorbent assay (ELISA) that permits direct determination of apoprotein (apo) CII, CIII, and E in total serum as well as in apo B-containing lipoprotein particles. To validate this ELISA technique, we studied several aspects of the assay: its specificity, the influence of the conditions of conservation of plasma and of lipoprotein fractions, the effect of delipidation, and its reproducibility. We measured the concentrations of apo CII, CIII, and E in total serum and in apo B-containing lipoproteins from a pool of normal sera and in sera from 75 healthy subjects. After sequential ultracentrifugation, the content of apo CII, CIII, and E in the major lipoprotein fractions was also determined. Total serum or plasma could be stored at -20 or -50 degrees C for at least six weeks and the isolated lipoprotein fractions for as long as four weeks, which suggests a protective effect of total serum on lipoprotein particle structure. Advantages of this ELISA include (a) its specificity, sensitivity, and reliability; (b) better discrimination than determination of total serum apoprotein; (c) easier application and greater rapidity; and (d) the possibility of application to population screening.

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