scholarly journals Produksi imunoglobulin Y (IgY) untuk pengembangan metode deteksi dini kontaminasi okratoksin (Immunoglobulin Y (IgY) production to develop an early detection method for ochratoxin contamination)

2018 ◽  
Vol 86 (1) ◽  
Author(s):  
Irma KRESNAWATY ◽  
. SUHARYANTO ◽  
. SISWANTO ◽  
Sumi HUDIYONO
Keyword(s):  
Early Detection ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Detection Method ◽  
Polyclonal Antibodies ◽  
Egg Laying ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Negative Effects ◽  
Sds Page

Indonesian coffee and cocoa commodities are constrained by low product quality problem due to contamination of fungal metabolites which  producing ochratoxin A (OTA). Ochratoxin is neprotoxic, immunogenic, carcinogenic and teratogenic to the human health. Early detection method on site detection should be developed  because of  those negative effects. The aim of this study  was to produce antibody to develop a method for  OTA detection. Antibody was produced by immunization of egg laying hen. Antibody-produced was sepatared and analyzed using ELISA (Enzyme-Linked Immunosorbent Assay) and DBIA (dot blot immunoassay),and tested its composition using HPLC and SDS PAGE. The results showed that anti-OTA polyclonal antibodies had been obtained already from chicken eggs in the 4th period (7 weeks after initial immunization). These antibodies showed anti-OTA reactivity by DBIA method and still showed anti-OTA reactivity up to 9th period (12 weeks after initial immunization). The anti-BSA antibodies produced should be removed to increase the sensitivity of antibodies againts ochratoxin A. The separation of BSA antibodies can be conducted by the absorption of the protein.  [Keywords: ochratoxin A; early detection; antibody IgY]. AbstrakKomoditas kopi dan kakao Indonesia terkendala masalah mutu produk yang rendah akibat kontaminasi cendawan penghasil okratoksin A. Okratoksin A (OTA) bersifat neprotoksik, imunogenik, karsinogenik dan teratogenik yang membahayakan kesehatan manusia. Karena efek negatif yang diakibatkan oleh mikotoksin ini, maka perlu dikembangkan deteksi dini kontaminasi okratoksin langsung di lokasi. Penelitian ini bertujuan menghasilkan antibodi imunoglobulin Y (IgY) untuk mengembangkan metode perakitan perangkat deteksi cepat berbasis imunologi untuk deteksi OTA. Antibodi dihasilkan menggunakan uji ayam petelur. Antibodi yang dihasilkan dipisahkan dan dianalisis aktivitasnya dengan ELISA (Enzyme-Linked Immunosorbent Assay) dan DBIA (dot blot immunoassay), serta diuji komposisinya dengan HPLC dan SDS PAGE. Hasil penelitian menunjukkan bahwa antibodi poliklonal anti-OTA sudah diperoleh dari telur ayam pada periode ke-4 (7 minggu setelah imunisasi awal). Antibodi ini menunjukkan reaktivitas anti-OTA dengan metode DBIA dan masih menunjukkan reaktivitas anti-OTA sampai periode 9 (12 minggu setelah imunisasi awal). Komposisi asam amino antibodi anti-OTA menunjukkan perbedaan dengan komposisi asam amino IgY di database. Antibodi anti BSA yang dihasilkan harus dihilangkan terlebih dahulu untuk meningkatkan sensitivitas antibodi terhadap okratoksin A dan pemisahan dapat dilakukan dengan penyerapan antibodi BSA.[Kata Kunci:  okratoksin A;  deteksi dini; antibodi IgY].

scholarly journals Immunodetection of Furcraea Necrotic Streak Virus-FSNV in Fique Plants (Furcraea Macrophylla Baker) Using A Polyclonal Antibody IgY Produced in Chicken Egg Yolk

2021 ◽  
Author(s):  
Deisy Toloza-Moreno ◽  
Laura Villamizar-Rivero ◽  
Paola Cuartas-Otálora ◽  
Gloria Barrera-Cubillos
Keyword(s):  
Early Detection ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Egg Yolk ◽  
Primary Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Concentration ◽  
Streak Virus ◽  
Dot Blot ◽  
Specificity And Sensitivity

Abstract The necrotic streak of the fique (Furcraea spp.) or “Macana” disease is considered the most limiting disease for this crop in Colombia, whose causal agent is the Furcraea Necrotic Streak Virus - FSNV (RNA +). Currently, there are no strategies to control the disease, being necessary to develop methods for early detection of this disease in the planting material before being taken to the field. In this study, polyclonal antibodies produced in egg yolk (IgY) were assesses for detection FSNV. Two immunoenzymatic methodologies were standardized: Dot Blot Immunobinding Assay (DBIA) and Enzyme Linked Immunosorbent Assay (ELISA), determining their specificity and sensitivity. The detection limit by DBIA corresponded to 8 µg/mL of purified virus suspension using 10 µg/mL of primary antibody. In the ELISA test, the primary antibody concentration of 3 µg/mL (1:800 dilution) detected the antigen at concentrations between 10 and 70 µg/mL. The polyclonal antibody anti-FNSV IgY allowed the detection of FNSV in samples of purified virus and extracts of roots and leaves of fique plants with symptoms of “Macana” disease and did not produce any signal with the control samples. Results showed the potential of using egg yolk IgY for immunological test as a novel approach for the early detection of FNSV in fique plants.

MYCOTOXINS IN BEANS OF PEANUTS GROWN IN TAJIKISTAN

2018 ◽  
Vol 1 (4) ◽  
pp. 74-77
Author(s):  
Sh. I. Razokov ◽  
◽  
D. M. Mirzoev ◽  
G. P. Kononenko ◽  
A. A. Burkin ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Cyclopiazonic Acid ◽  
Practical Significance ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Systems ◽  
The Republic ◽  
Combined Contamination

The article presents the results of an extensive mycotoxicological examination of 11 samples of peanut beans grown in two regions of the Republic of Tajikistan. The determination of 16 mycotoxins was carried out by indirect competitive enzyme-linked immunosorbent assay using commercial and certified research test systems. It has been established that for peanut beans in this area, a combined contamination by a group of sanitary-significant mycotoxins, including diacetoxyscirpenol, alternariol, ochratoxin A, PR-toxin and cyclopiazonic acid, is characteristic. The prospects of further research and the practical significance of the results are discussed.

Development of a quantitative and sensitive enzyme-linked immunosorbent assay for ochratoxin A using antibodies from the yolk of the laying hen

1993 ◽  
Vol 41 (10) ◽  
pp. 1784-1789 ◽  
Author(s):  
J. R. Clarke ◽  
R. R. Marquardt ◽  
A. Oosterveld ◽  
A. A. Frohlich ◽  
F. J. Madrid ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Laying Hen ◽  
Enzyme Linked Immunosorbent Assay

scholarly journals Mycotoxins as a risk in the grain food

2009 ◽  
pp. 79-86 ◽  
Author(s):  
Jovana Matic ◽  
Jasna Mastilovic ◽  
Ivana Cabarkapa ◽  
Anamarija Mandic
Keyword(s):  
European Union ◽  
Secondary Metabolites ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Toxic Effects ◽  
Enzyme Linked Immunosorbent Assay ◽  
The European Union ◽  
The Mean ◽  
Grain Foods

Mycotoxins are toxic secondary metabolites of fungi that contaminate a large variety of foods and have toxic effects on humans. The best protection against mycotoxins is to monitor their presence in food. This paper shows the screening results of mycotoxins present in 76 samples of different groups of grain foods. Samples of grain food were analyzed for contamination with aflatoxins, ochratoxin A, zearalenone, fumonisins and deoxynivalenol. Analysis were conducted using competitive enzyme-linked immunosorbent assay (ELISA). None of the samples was contaminated with aflatoxins. The most predominant mycotoxin was ochratoxin A with the mean level of 4.84 ? 4.49 ppb in 19.7% of the examined samples. Zearalenone, fumonisins, and deoxynivalenol were found in 9.21, 14.5 and 3.9% of the samples, respectively. Mycotoxin content in the investigated samples was compared with the regulations of Serbia and those of the European Union.

scholarly journals Aflatoxigenic and ochratoxigenic moulds and their toxins in tree nuts on Serbian market

2020 ◽  
pp. 9-18
Author(s):  
Irena Rakic ◽  
Gordana Dimic ◽  
Marija Skrinjar ◽  
Suncica Kocic-Tanackov
Keyword(s):  
Aspergillus Niger ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Alternaria Alternata ◽  
Rhizopus Oryzae ◽  
Limit Of Detection ◽  
Average Concentration ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mean Values ◽  
Tree Nuts

In this study, moulds and mycotoxins presence in different tree nuts were investigated. The results showed that all of the 25 samples were contaminated with moulds. Mean values of total mould count varied from 1-4.9 cfu per grain. The most frequent species in hazelnut samples were Rhizopus oryzae (32.2%) and Aspergillus niger (28.9%). In walnuts A. niger (75.6%), in cashews also A. niger (42.4%) while in pistachio samples Alternaria alternata (20.7%), and Cladosporium cladosporioides (20.7%) were the most dominant. Rhizopus oligosporus was the only identified species in all almond samples (100%). Using Enzyme Linked Immunosorbent Assay (ELISA), the presence of total aflatoxins and ochratoxin A was examinated. In all analyzed samples, levels of ochratoxin A were below the limit of detection. Total aflatoxins were detected only in walnut samples with average concentration of 7.1 ?g/kg.

scholarly journals Levels of Airborne Soybean Allergen (Gly m 1) in a Brazilian Soybean Production City: A Pilot Study

2020 ◽  
Vol 17 (15) ◽  
pp. 5381
Author(s):  
Cinthya Covessi Thom de Souza ◽  
Nelson Augusto Rosário Filho ◽  
Juliana Francis de Camargo ◽  
Ricardo Henrique Moreton Godoi
Keyword(s):  
Pilot Study ◽  
Immunosorbent Assay ◽  
Detection Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Soybean Production ◽  
Soy Production ◽  
Assay Detection ◽  
Soybean Allergen

Asthma epidemics have been shown to be related to where soybeans are loaded and handled, but data are scarce in the literature. This pilot study evaluated the levels of Gly m 1 in dust samples collected in Maringá, Brazil, a city with high soy production and processing. A dust impactor was used to collect seven isolated samples during 2015 and 2016. Samples were analyzed by an ELISA (enzyme-linked immunosorbent assay) detection method. Gly m 1 was found in all samples, ranging from 0.82–24.38 ng/m3 (median 2.41), regardless of the month or year evaluated. The levels of Gly m 1 were considered low, but the concentrations required to cause sensitization and symptoms are uncertain.

scholarly journals Enzyme-Linked Immunosorbent Assay for Simultaneous Detection of Two Fungicides Kresoxim-methyl and Trifloxystrobin in Oranges

2016 ◽  
Vol 34 (No. 5) ◽  
pp. 429-438 ◽  
Author(s):  
Yang Wei ◽  
Zhu Jie ◽  
Liu Ming-Qi ◽  
Dai Xian-Jun
Keyword(s):  
Immunosorbent Assay ◽  
Reaction Rate ◽  
Polyclonal Antibodies ◽  
Simultaneous Detection ◽  
Acid Methyl Ester ◽  
Enzyme Linked Immunosorbent Assay ◽  
Strobilurin Fungicides ◽  
Acid Methyl ◽  
Recovery Study ◽  
The Common

To assemble an indirect competitive enzyme-linked immunosorbent assay (ELISA) for estimating two strobilurin fungicides at the same time, the hapten was synthesised which contained the common active group, (E)-2-(2-bromo-phenyl)-2-(methoxyimino) acetic acid methyl ester (OEBr) in kresoxim-methyl and trifloxystrobin. The immunogen and coating antigen were respectively prepared through conjugating the above-mentioned hapten with BSA and OVA by the mixed anhydride and activated ester methods, and polyclonal antibodies were produced by immunised rabbits. An enzyme-linked immunosorbent assay was developed for simultaneous detection of kresoxim-methyl and trifloxystrobin. In ELISA, the antiserum showed high affinity and sensitivity to kresoxim-methyl and trifloxystrobin, and their IC<sub>50</sub> value and detection limit (expressed as IC<sub>10</sub>) were 14.7 and 0.0044 µg/ml, respectively, for kresoxim-methyl, and 22.9 and 0.014 µg/ml, respectively for trifloxystrobin. The cross-reaction rate was below 0.1% for other strobilurin fungicides. Recovery study of ELISA from spiked samples of homogenised peeled oranges (final concentrations of 100, 10, and 1 µg/ml) resulted in recovery levels in the range of 82–104%.

Quantification of serum amyloid P by enzyme-linked immunosorbent assay

1991 ◽  
Vol 37 (10) ◽  
pp. 1742-1745 ◽  
Author(s):  
R Saïle ◽  
A Kandoussi ◽  
M Deveaux ◽  
J Descamps ◽  
E Hachulla ◽  
...  
Keyword(s):  
Human Plasma ◽  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Serum Amyloid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Amyloid P ◽  
Assay Procedure ◽  
Normal Individuals ◽  
Sandwich Type ◽  
Amyloid P

Abstract This enzyme-linked immunosorbent assay procedure for quantifying serum amyloid P (SAP) in human plasma makes use of affinity-purified polyclonal antibodies to SAP in a "sandwich"-type format. The procedure is sensitive, reproducible, simple, and easily automatable. Results correlate well with those by a rocket immunoelectrophoresis method performed with the same antibodies. Sera from apparently normal individuals had a mean SAP content of 44.17 mg/L and increased with age.

Development of a Highly Sensitive Enzyme-Linked Immunosorbent Assay Based on Polyclonal Antibodies for the Detection of Polychlorinated Dibenzo-p-dioxins

1998 ◽  
Vol 70 (6) ◽  
pp. 1092-1099 ◽  
Author(s):  
Yukio Sugawara ◽  
Shirley J. Gee ◽  
James R. Sanborn ◽  
S. Douglass Gilman ◽  
Bruce D. Hammock
Keyword(s):  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Highly Sensitive
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