Rhizobium strain identification in Arachis hypogaea nodules by enzyme-linked immunosorbent assay (ELISA)

1978 ◽  
Vol 24 (12) ◽  
pp. 1537-1543 ◽  
Author(s):  
B. Kishinevsky ◽  
M. Bar-Joseph
Keyword(s):  
Arachis Hypogaea ◽  
Immunosorbent Assay ◽  
Root Nodules ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Strain Identification ◽  
Single Root ◽  
Rhizobium Strains ◽  
Serological Identification ◽  
Phosphate Buffered Saline

The technique of enzyme-linked immunosorbent assay (ELISA) was used for serological identification of peanut Rhizobium strains both in cell suspension of pure culture and in single root nodules of groundnut (Arachis hypogaea) plants. Antisera of three peanut Rhizobium strains were tested against eight different Rhizobium isolates. Three serogroups identified by agglutination and immunodiffusion tests were confirmed by ELISA.In this experiment ELISA was more sensitive by four to six orders of magnitude than the agglutination and immunodiffusion tests and enabled the detection of Rhizobium antigens in cell suspensions of 104–105 cells per millilitre.The reactions of culture and nodule antigens were identical for all strains investigated.ELISA enabled the precise typing of rhizobial isolates in single small root nodules. The minimum fresh weight of nodule tissue necessary to perform the ELISA test was 0.4 mg crushed in 1 ml of phosphate-buffered saline (PBS).ELISA was also successfully used for strain identification in mixed inoculated plants. One of the strains in each pair formed most of the nodules examined.

Studies on the enzyme linked immunosorbent assay (ELISA) test forSchistosoma mansoniinfections

1978 ◽  
Vol 72 (3) ◽  
pp. 243-253 ◽  
Author(s):  
M. McLaren ◽  
C. C. Draper ◽  
J. M. Roberts ◽  
E. Minter-Goedbloed ◽  
G. S. Ligthart ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay

Competitive Direct Enzyme-Linked Immunosorbent Assay for Detection of Sulfamethazine Residues in Swine Urine and Muscle Tissue

1988 ◽  
Vol 71 (6) ◽  
pp. 1137-1140 ◽  
Author(s):  
Deborah E Dixon-Holland ◽  
Stanley E Katz
Keyword(s):  
Horseradish Peroxidase ◽  
Muscle Tissue ◽  
Immunosorbent Assay ◽  
Microtiter Plate ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sensitive Assay ◽  
Saline Extract ◽  
Swine Urine ◽  
Phosphate Buffered Saline

Abstract A sensitive assay for the detection of sulfamethazine in swine urine and muscle tissue using a direct competitive enzyme-linked immunosorbent assay (ELISA) has been developed. Undiluted urine or a phosphate-buffered saline extract of pork muscle tissue is mixed with an enzyme-labeled conjugate of sulfamethazine and horseradish peroxidase. The mixture is added to wells of a microtiter plate coated with antibody to sulfamethazine. After the test system is incubated, washed, and re-incubated with substrate and the reaction is stopped, the absorbance is measured at 405 nm. Levels of sulfamethazine as low as 20 ng sulfamethazine/g muscle tissue and 10 ng sulfamethazine/ mL swine urine were detected and estimated

Enzyme-Linked Immunosorbent Assay and Microbiologic Culture for Diagnosis of Staphylococcus aureus Intramammary Infection in Cows

1996 ◽  
Vol 59 (1) ◽  
pp. 6-10 ◽  
Author(s):  
PAUL C. BARTLETT ◽  
RONALD J. ERSKINE ◽  
PATRICK GASTON ◽  
PHILIP M. SEARS ◽  
HENDRICUS WILHELMUS HOUDIJK
Keyword(s):  
Staphylococcus Aureus ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Relative Sensitivity ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infection Status ◽  
Intramammary Infection ◽  
Previous Infection ◽  
Current Infection

Recent reports have indicated that the relative sensitivity and specificity of the ELISA test for detection of intramammary infection of cows with Staphylococcus aureus is not as high as originally reported. It has been suggested that antibodies measured by enzyme-linked immunosorbent assay (ELISA) more closely reflect previous infection status rather than current infection status, and that the delay in antibody formation following infection and the persistence of antibodies after elimination of infection may be responsible for some of the discrepancy observed between ELISA and bacterial culture results conducted on the same milk sample. This study (n = 209 cows) was undertaken to determine if an ELISA for S. aureus intramammary infection more closely reflects previous infection status than it does current infection status, and to ascertain whether correction of this time-delay factor substantially improves calculated values of ELISA relative sensitivity and specificity. Receiver-operator curves were constructed to compare different time-related definitions of microbiologic culture results used for comparison with ELISA results. A greater degree of curvature in receiver-operator curves indicated that ELISA results did more closely reflect culture results performed on milk samples taken 1 and 3 weeks previously. Insignificant improvement in sensitivity and specificity occurred when the database was limited to cows (n = 140) with milk production greater than 13.6 kg/day. However, values of sensitivity were all less than or equal to 90%, and values of specificity were all less than 54%.

scholarly journals Antibody Responses to CampylobacterInfections Determined by an Enzyme-Linked Immunosorbent Assay: 2-Year Follow-Up Study of 210 Patients

2001 ◽  
Vol 8 (2) ◽  
pp. 314-319 ◽  
Author(s):  
Mette Aagaard Strid ◽  
Jørgen Engberg ◽  
Lena Brandt Larsen ◽  
Kamilla Begtrup ◽  
Kåre Mølbak ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Serum Antibody ◽  
Large Degree ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Campylobacter Coli ◽  
Negative Results ◽  
Heat Stable Antigen

ABSTRACT An enzyme-linked immunosorbent assay (ELISA) was adapted to measure immunoglobulin G (IgG), IgM, and IgA classes of human serum antibody toCampylobacter jejuni and Campylobacter coli. Heat-stable antigen, a combination of C. jejuni serotype O:1,44 and O:53 in the ratio 1:1, was used as a coating antigen in the ELISA test. A total of 631 sera from 210 patients with verifiedCampylobacter enteritis were examined at various intervals after infection, and a control group of 164 sera were tested to determine the cut-off for negative results. With a 90th percentile of specificity, IgG, IgM, and IgA showed a sensitivity of 71, 60, and 80%, respectively. By combining all three antibody classes, the sensitivity was 92% within 35 days after infection, whereas within 90 days after infection, a combined sensitivity of 90% was found (IgG 68%, IgM 52%, and IgA 76%). At follow-up of the patients, IgG antibodies were elevated 4.5 months after infection but exhibited a large degree of variation in antibody decay profiles. IgA and IgM antibodies were elevated during the acute phase of infection (up to 2 months from onset of infection). The antibody response did not depend on Campylobacter species or C. jejuniserotype, with the important exception of response to C. jejuni O:19, the serotype most frequently associated with Guillain-Barré syndrome. All of the patients infected with this serotype had higher levels of both IgM (P = 0.006) and IgA (P = 0.06) compared with other C. jejuni and C. coli serotypes.

scholarly journals Vaccination in Nile tilapia broodstock with whole cell vaccine and disease resistance in its fry against Aeromonas hydrophila

2017 ◽  
Vol 16 (2) ◽  
pp. 268 ◽  
Author(s):  
Sukenda Sukenda ◽  
Odang Carman ◽  
Rahman Rahman ◽  
Dendi Hidayatullah ◽  
Nurfitriani Siti Yumaidawati
Keyword(s):  
Disease Resistance ◽  
Immunosorbent Assay ◽  
Antibody Level ◽  
Aeromonas Hydrophila ◽  
Nile Tilapia ◽  
Cell Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Whole Cell ◽  
Whole Cell Vaccine ◽  
Phosphate Buffered Saline

<p class="NoParagraphStyle"><strong>ABSTRACT</strong></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle">The aim of this study was to analyze the effectivity of vaccination in Nile tilapia broodstock with whole cell vaccine and disease resistance in fry tilapia against <em>Aeromonas hydrophila</em>. Tilapia Nirwana strain that used for this had average body weight of 185±13.23 g and were maintained in ponds sizing of (2.5×2.5×1 m<sup>3</sup>). Vaccinations that has been done through intraperitoneal injection using dose of 0.1 mL/fish, meanwhile the fish for control was injected by phosphate buffered saline (PBS). This study used complete randomized design with two treatments and three replications. Antibody level was measured by using indirect enzyme-linked immunosorbent assay (ELISA) method in the broodstock, egg, and fry.  Challenge test in fry tilapia performed at the age of 5, 10, and 15 days. The results showed that vaccination in tilapia broodstock delivered a significant antibody level in broodstock, eggs, and fry (P&lt;0.05) compared to the control. Relative percent survival of offspring at 5, 10, and 15 days were 78.26%, 70.59%, and 65.52%, respectively.  As a conclusion, vaccination in tilapia broodstock was effective to improve specific and non-specific immunity, and protect fry tilapia from <em>A. hydrophila</em> infection through maternal immunity.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: vaccination, antibody, maternal immunity, tilapia, <em>Aeromonas hydrophila</em></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle"><strong>ABSTRAK</strong><strong></strong></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle">Penelitian ini bertujuan untuk menganalisis efikasi vaksinasi pada induk nila dengan vaksin sel utuh dan ketahanan benih yang dihasilkan terhadap <em>Aeromonas hydrophila</em>. Ikan nila stain Nirwana yang digunakan dalam penelitian memiliki bobot rata-rata 185±13,23 g dan ikan dipelihara dalam kolam (2,5×2,5×1 m<sup>3</sup>). vaksinasi dilakukan melalui penyuntikan intraperitoneal dengan dosis 0,1 mL/ikan, sementara itu ikan kontrol disuntik dengan <em>phosphate buffered saline</em> (PBS). Penelitian ini menggunakan rancangan acak lengkap dengan dua perlakuan dan tiga ulangan. Tingkat antibodi diukur dengan menggunakan metode<em> indirect enzyme-linked immunosorbent assay</em> (ELISA) pada induk, telur dan benih. Uji tantang pada benih dilakukan pada umur 5, 10, dan 15 hari. Hasil penelitian menunjukan bahwa vaksinasi pada induk nila secara signifikan dapat meningkatkan level antibodi pada induk, telur, dan benih (P&lt;0,05) dibandingkan dengan kontrol. Kelangsungan hidup relatif pada benih berumur 5, 10, dan 15 hari masing-masing adalah 78,26%; 70,59%; dan 65,52%. Sebagai kesimpulan vaksinasi pada induk nila efektif dalam memperbaiki imunitas spesifik dan non spesifik serta melindungi benih dari infeksi <em>A. hydrophila</em> melalui imunitas maternal.</p><p class="NoParagraphStyle"> </p><p>Kata kunci: vaksinasi, antibodi, imunitas maternal, ikan nila, <em>Aeromonas hydrophila</em></p>

scholarly journals Epidémiologie de la fièvre Hémorragique de Crimée-Congo (FHCC) chez les bovins dans le département de Boboye au Niger

2020 ◽  
Vol 14 (3) ◽  
pp. 698-705
Author(s):  
Alima Maïna ◽  
Abdoulkarim Issa Ibrahim ◽  
Abdou Alassane ◽  
Hassane Adakal
Keyword(s):  
Immunosorbent Assay ◽  
Disease Transmission ◽  
Local Governments ◽  
Indirect Elisa ◽  
Hemorrhagic Fever ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Crimean Congo Hemorrhagic Fever ◽  
Cchf Virus ◽  
Made In

La distribution et la dynamique des populations des tiques est un élément clé dans la connaissance des maladies transmises par ces vecteurs. C’est ainsi que cette étude a été conduite afin de mieux connaître l’épidémiologie de la Fièvre Hémorragique de Crimée-Congo (FHCC) dans les 8 communes du département de Boboye au Niger, où 355 sérums de bovins ont été collectés. En plus des sérums, des tiques ont été collectées sur 144 bovins, soit 18 par commune. Les sérums ont été soumis à un test ELISA (Enzyme Linked Immunosorbent Assay) indirect pour la détection d’anticorps anti-FHCC. Soixante-douze (72) éleveurs ont été interviewés sur leur connaissance de l’écologie des tiques, vecteurs du virus de la FHCC. Les résultats de l’enquête ont révélé que les éleveurs n’ont pas recours aux acaricides et que, dans leur majorité (55/72 soit 76,4 %), ils pratiquent la transhumance. L’étude a permis l’identification de 1342 tiques réparties en trois genres : Hyalomma (91,7%), Amblyomma (5,7%) et Rhipicephalus (Boophilus) (2,6%). La séroprévalence globale a été de 9,1±0,03%. Les communes de Harikanassou et Kiota ont été celles où les fortes prévalences ont été observées de 26,7 ± 12,9% et 22,5 ±12,9%. Le virus de la FHCC est en circulation chez la population animale, alors des investigations doivent être faites chez la population humaine.Mots clés : Anticorps anti-FHCC, Enzyme Linked Immunosorbent Assay Indirecte, Prévalence, Sérums, Tiques.   English Title: Crimean-Congo Hemorrhagic Fever (CCHF) ’s Epidemiology in cattle in Boboye’s department of Niger Republic To understand disease transmission by ticks, knowledge of population dynamics and distribution of these vectors are essentials. To sought that, the epidemiology of Crimean-Congo Hemorrhagic Fever (CCHF) in Niger Republic was studied by sampling 355 bovines (sera and ticks) in eight (8) local governments in Boboye’s department. Eighteen (18) bovines were sampled for ticks collection per local government making them a total of 144 bovine. Indirect ELISA test (enzyme-linked immunosorbent assay) was used to detect anti- CCHF antibodies. Seventy-two (72) farmers were surveyed on their knowledge on ticks’ ecology, main vectors of CCHF virus. The results revealed that farmers are not using acaricides, and their majority (55/72 thus 76.4%) practice Transhumance. The study allowed the identification of 1342 ticks distributed in 3 genus: Hyalomma (91.7%), Amblyomma (5.7%) and Rhipicephalus (Boophilus) (2.6%). The global seroprevalence against CCHF was (9.1 ± 0.03) %. Harikanassou and Kiota were the most affected local governments with respectively (26.7±12.9) % and (22.5±12,9) % prevalence. CCHV virus is circulating in animal population, so investigations must be made in human population. Keywords: Anti-CCHF antibodies, Indirect Enzyme Linked Immunosorbent Assay, Prevalence, Sera, Ticks.

Validated Sandwich Enzyme-Linked Immunosorbent Assay for Casein and Its Application to Retail and Milk-Allergic Complaint Foods

2004 ◽  
Vol 67 (9) ◽  
pp. 1933-1938 ◽  
Author(s):  
SUSAN L. HEFLE ◽  
DEBRA M. LAMBRECHT
Keyword(s):  
Immunosorbent Assay ◽  
Milk Proteins ◽  
Food Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Food Ingredients ◽  
Milk Contamination ◽  
Quality Control Tool ◽  
Sandwich Type ◽  
Life Threatening ◽  
Phosphate Buffered Saline

Cows' milk is a commonly allergenic food. Cross-contamination of milk proteins into nondairy, kosher-pareve foods prepared on shared processing equipment can cause severe, life-threatening reactions in milk-allergic individuals. A sandwich-type enzyme-linked immunosorbent assay (ELISA; 96-well plate format) was developed for the detection of undeclared casein in foods. Rabbit anti-casein antibodies were used as the capture reagent. Food samples and standards were ground, extracted in 0.01 M phosphate-buffered saline, clarified by centrifugation, and added to the wells. Goat anti-casein antibodies were employed as the detector antibody, and the amount of antibody bound was determined with a commercial rabbit anti-goat immunoglobulin conjugated to alkaline phosphatase, with subsequent substrate reaction. Antibodies developed were specific to casein, with no cross-reaction observed with 30 foods and food ingredients. Non–milk-containing products such as fruit juices, fruit juice bars, sorbets, and dark and pareve-labeled chocolate were purchased from June 2002 through June 2003. In addition, samples allegedly causing eight milk-allergic consumer complaints were analyzed. The ELISA had a detection limit of less than 0.5 ppm of casein. The casein content in the analyzed foods ranged from less than 0.5 ppm to more than 40,000 ppm casein; undeclared casein residues were found in all of the samples implicated in allergic reactions. The levels of milk contamination in some of the other surveyed products could also be hazardous for milk-allergic consumers. This ELISA method provides a useful quality control tool for the food industry and could also be used as a validation of kosher-pareve status.

scholarly journals Indirect Enzyme Linked Immunosorbent Assay Sebagai Metode untuk Melacak Bruselosis pada Sapi Perah (INDIRECT ENZYME IMMUNOSORBENT ASSAY (I-ELISA) AS METHOD FOR DETECT BRUCELLOSIS IN DAIRY COW)

2019 ◽  
Vol 20 (1) ◽  
pp. 30
Author(s):  
Rinaldi Ghurafa ◽  
Denny Widaya Lukman ◽  
Hadri Latif
Keyword(s):  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
Kappa Value ◽  
Alternative Test ◽  
The Rose ◽  
Higher Sensitivity ◽  
Good Agreement

Brucellosis has become a zoonotic disease that received attention in efforts to prevent and eradicate strategic infectious animal diseases in Indonesia. Brucellosis can be detected early by the rose bengal test (RBT), followed by complement fixation test (CFT) and by enzyme linked immunosorbent assay (ELISA). The aims of this research was to study the indirect enzyme linked immunosorbent assay test (I-ELISA) as an alternative test for detecting brucellosis in dairy cattle. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated sensitivity and specificity, as well as analyzed by calculating the kappa value. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated for sensitivity and specificity, as well as analyzed by calculating the Kappa statistical value. The results of the sensitivity and specificity calculation showed that the indirect enzyme linked immunosorbent assay (I-ELISA) test developed a higher sensitivity (100%) compared to RBT test (93.75%) and commercial I-ELISA (93.75%). The developed I-ELISA specificity (74.68%) was still lower than RBT (89.87%), but higher than commercial I-ELISA (70.52%). The calculation of the statistical value of kappa RBT with CFT showed the kappa value 0.7120 which meaned it had a good agreement, commercial I-ELISA with CFT showed kappa value 0.6165 which meaned it had good suitability, whereas I-ELISA developed with CFT showed kappa value 0.4984 which meaned having a moderate agreement.In conclusion, the indirect enzyme linked immunosorbent assay (I-ELISA) which had been developed had low specificity, but the sensitivity was the highest compared to the commercial I-ELISA test and RBT, so this test was appropriate to be used as a screening test, especially in dairy cows movement into brucellosis-free areas or regions.

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