scholarly journals Evaluation of an Indirect ELISA for the Diagnosis of Bovine Brucellosis

1995 ◽  
Vol 7 (4) ◽  
pp. 473-475 ◽  
Author(s):  
Francisco A. Uzal ◽  
Alejandra E. Carrasco ◽  
Susana Echaide ◽  
Klaus Nielsen ◽  
Carlos A. Robles
Keyword(s):  
Indirect Elisa ◽  
Screening Tests ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Brucellosis ◽  
Energy Agency ◽  
Elisa Kit ◽  
Large Numbers ◽  
The Rose ◽  
Epidemiological Information ◽  
Control And Eradication

Control and eradication of bovine brucellosis is usually based on the serological detection of antibodies. In Argentina, the rose bengal test (RB) and the buffered plate antigen test (BPA) are the 2 screening tests officially recognized, whereas the 2-mercaptoethanol test (2ME) and the tube agglutination test (SAT) are the confirmatory assays currently in use. In order to improve the serological diagnosis of bovine brucellosis in Patagonia, Argentina, an indirect enzyme-linked immunosorbent assay (I-ELISA) kit distributed by the International Atomic Energy Agency was evaluated. Sera from negative nonvaccinated, negative but vaccinated, and positive animals were tested by all the above techniques. The specificity of the I-ELISA (98.9% and 97.2%) was similar to that of the BPA, RB, 2ME and complement fixation (CF) tests when used to test sera from negative nonvaccinated and negative but vaccinated animals, respectively. The sensitivity of the I-ELISA (98.7%) was higher than the BPA test (96.1%) and the CF test (95.2%). The I-ELISA kit evaluated in this study was thought to be a valuable tool for the diagnosis of bovine brucellosis in the Patagonia region where little epidemiological information is available about this disease, and where large numbers of sera should be tested to obtain such information.

scholarly journals Comparative Performance of In-House and Commercially Available ELISA Kits for the Detection of Antibody Against Newcastle Disease Virus Vaccine

2013 ◽  
Vol 22 (1-2) ◽  
pp. 55-64
Author(s):  
MMI Chowdhury ◽  
MT Islam ◽  
A Aktar ◽  
MKJ Bhuiyan ◽  
MM Kamal ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Sensitivity And Specificity ◽  
Newcastle Disease ◽  
Indirect Elisa ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Dilution ◽  
Positive Correlation ◽  
Elisa Kit

An In-house Indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of serum antibody titre against Newcastle disease virus (NDV) and was compared its sensitivity and specificity with the commercially available NDV antibody detection ELISA kit (Biocheck®, USA). The reference NDV was purified by centrifugation, ultracentrifugation and by sucrose density gradient ultracentrifugation. This purified NDV was used for coating of 96-well flat bottomed microtitre plate and to raise hyperimmune sera (known) in Fayoumi chickens. In the standardization test, the antigen dilution of 10-6 and the serum dilution of 10-3 were considered to be optimum for the present ELISA system. The correlation regression analysis was performed to construct a standard curve equation where a good positive correlation was observed (r = 0.912, n = 8, P<0.01). The equation was used to convert corrected absorbance readings of the single working dilution (1 : 1000) directly into predicted ELISA antibody activity titres. In the sensitivity and specificity test, the serum dilution of 10-5 appeared to be the highest dilution which had the maximum lowest capacity to bind with the coated antigen of the present ELISA kit and only anti-NDV serum was found to bind with the coated antigen instead of serum of IBDV in the plate which revealed the high specificity of the developed In-house Indirect ELISA kit. In a comparative study with the 80 chicken sera samples, a significant positive correlation (r = 0.901, n = 80, P<0.01) was found between In-house Indirect ELISA and commercial ELISA kit (Biocheck®, USA).DOI: http://dx.doi.org/10.3329/pa.v22i1-2.16467 Progress. Agric. 22(1 & 2): 55 - 64, 2011

scholarly journals Brucella abortus S19 GFP-tagged vaccine allows the serological identification of vaccinated cattle

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0260288
Author(s):  
Carlos Chacón-Díaz ◽  
Ana Zabalza-Baranguá ◽  
Beatriz San Román ◽  
José-María Blasco ◽  
Maite Iriarte ◽  
...  
Keyword(s):  
Brucella Abortus ◽  
Fluorescent Protein ◽  
Indirect Elisa ◽  
Economic Losses ◽  
Bovine Brucellosis ◽  
Serological Identification ◽  
Green Fluorescent ◽  
Chromosome Ii ◽  
The Rose ◽  
S19 Vaccine

Bovine brucellosis induces abortion in cows, produces important economic losses, and causes a widely distributed zoonosis. Its eradication was achieved in several countries after sustained vaccination with the live attenuated Brucella abortus S19 vaccine, in combination with the slaughtering of serologically positive animals. S19 induces antibodies against the smooth lipopolysaccharide (S-LPS), making difficult the differentiation of infected from vaccinated bovines. We developed an S19 strain constitutively expressing the green fluorescent protein (S19-GFP) coded in chromosome II. The S19-GFP displays similar biological characteristics and immunogenic and protective efficacies in mice to the parental S19 strain. S19-GFP can be distinguished from S19 and B. abortus field strains by fluorescence and multiplex PCR. Twenty-five heifers were vaccinated withS19-GFP (5×109 CFU) by the subcutaneous or conjunctival routes and some boosted with GFP seven weeks thereafter. Immunized animals were followed up for over three years and tested for anti-S-LPS antibodies by both the Rose Bengal test and a competitive ELISA. Anti-GFP antibodies were detected by an indirect ELISA and Western blotting. In most cases, anti-S-LPS antibodies preceded for several weeks those against GFP. The anti-GFP antibody response was higher in the GFP boosted than in the non-boosted animals. In all cases, the anti-GFP antibodies persisted longer, or at least as long, as those against S-LPS. The drawbacks and potential advantages of using the S19-GFP vaccine for identifying vaccinated animals in infected environments are discussed.

scholarly journals Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS-CoV-2 seroconversion

2021 ◽  
Author(s):  
Marcelo S. Conzentino ◽  
Karl Forchhammer ◽  
Emanuel M Souza ◽  
Fábio O. Pedrosa ◽  
Meri B. Nogueira ◽  
...  
Keyword(s):  
Mass Production ◽  
Nucleocapsid Protein ◽  
Indirect Elisa ◽  
Humoral Response ◽  
High Yield ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Assay Sensitivity ◽  
Elisa Kit ◽  
Chromatography Step

AbstractSerological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here the SARS-CoV-2 nucleocapsid protein was expressed in Escherichia coli and purified to homogeneity and high yield using a single chromatography step. The purified SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identification of human SARS-CoV-2 seroconverts. The assay sensitivity and specificity were determined analyzing sera from 140 PCR-confirmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specificity; identical accuracies were obtained in head-to-head comparison with a commercial ELISA kit. Antigen coated plates were stable for up to 3 months at 4°C). The ELISA method described is ready to mass production and will be an additional toll to track COVID-19 cases.

scholarly journals Estimating the probability of freedom from bovine brucellosis in the Galapagos Islands

2018 ◽  
Vol 147 ◽  
Author(s):  
G. Gioia ◽  
R. L. Vinueza ◽  
M. Cruz ◽  
M. Jay ◽  
Y. Corde ◽  
...  
Keyword(s):  
Latin America ◽  
Indirect Elisa ◽  
Galapagos Islands ◽  
Galápagos Islands ◽  
Bovine Brucellosis ◽  
Santa Cruz ◽  
Negative Results ◽  
Level Design ◽  
Cattle Farms ◽  
The Rose

AbstractBovine brucellosis is a worldwide zoonotic disease that still burdens several countries in the Mediterranean, Asia, Africa and Latin America. Although the disease is present in Ecuador, the Galapagos Islands seem to be free from the disease based on a survey conducted in 1997 where all tested animals showed negative results. This study aimed at estimating the probability of freedom from brucellosis in this Ecuadorian province in 2014. A survey was implemented on the three main cattle-producing islands of the province: Santa Cruz, Isabela and San Cristóbal. Thirty-three cattle farms and 410 cattle were tested for brucellosis using the Rose Bengal test and indirect ELISA. All animals showed negative results for both tests. Probability of freedom was estimated at 98%, 91% and 88% for Santa Cruz, Isabela and San Cristóbal, respectively, considering a herd-level design seroprevalence of 20% and animal-level design seroprevalence of 15%, and assuming a perfect specificity of the survey. The negative results found in 1997 and present surveys suggest that the Galapagos Islands are free from bovine brucellosis.

scholarly journals Serological investigation of bovine brucellosis in three cattle production systems in Yewa Division, south-western Nigeria

2013 ◽  
Vol 84 (1) ◽  
Author(s):  
Simeon I.B. Cadmus ◽  
Peter I. Alabi ◽  
Hezekiah K. Adesokan ◽  
Emma J. Dale ◽  
Judy A. Stack
Keyword(s):  
Production Systems ◽  
Herd Size ◽  
Cross Sectional Study ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Brucellosis ◽  
Cattle Production ◽  
Serological Screening ◽  
Cross Sectional ◽  
Ogun State ◽  
The Rose

Limited data are available on the risk factors responsible for the occurrence of brucellosis amongst different cattle production systems in Nigeria despite its significant impact on livestock production. Consequently, a cross-sectional study was conducted to determine the prevalence of bovine brucellosis in three cattle production systems in Yewa Division of Ogun State, south-western Nigeria. A total of 279 blood samples (sedentary = 88; transhumance = 64; trade = 127) were examined for antibodies to Brucella sp. using the Rose Bengal test (RBT) and competitive enzyme-linked immunosorbent assay (cELISA). Overall, 24 (8.6%) and 16 (5.7%) of the animals tested seropositive for Brucella using RBT and cELISA, respectively. The herd seroprevalences based on RBT and cELISA were 31.6% and 15.8%, respectively. The results using cELISA reveal higher seroprevalence in the trade cattle (7.9%; confidence intervals [CI] = 3.2% – 12.6%) and those in a sedentary system (5.7%; CI = 0.9% – 10.5%) than in cattle kept under a transhumant management system (1.6%; CI = 1.5% – 4.7%). Age (> 3 years; p = 0.043) and breed (Djali; p = 0.038) were statistically significant for seropositivity to brucellosis based on cELISA, but sex (female, p = 0.234), production system (trade and sedentary; p = 0.208) or herd size (> 120; p = 0.359) was not. Since breeding stock is mostly sourced from trade and sedentary cattle, it is important that routine serological screening should be conducted before introducing any animal into an existing herd.

D-Dimer Determination to Exclude Pulmonary Embolism: a Two-Step Approach Using Latex Assay as a Screening Tool

1994 ◽  
Vol 72 (01) ◽  
pp. 089-091 ◽  
Author(s):  
P de Moerloose ◽  
Ph Minazio ◽  
G Reber ◽  
A Perrier ◽  
H Bounameaux
Keyword(s):  
Pulmonary Embolism ◽  
Screening Tool ◽  
Screening Tests ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Elisa Technique ◽  
Diagnostic Step ◽  
D Dimer ◽  
Rule Out ◽  
Latex Test

SummaryD-dimer (DD), when measured by a quantitative enzyme-linked immunosorbent assay (ELISA), is a valuable test to exclude venous thromboembolism (VTE). However, DD ELISA technique is not appropriate for emergency use and the available agglutination latex assays are not sensitive enough to be used as an alternative to rule out the diagnosis of VTE. Latex assays could still be used as screening tests. We tested this hypothesis by comparing DD levels measured by ELISA and latex assays in 334 patients suspected of pulmonary embolism. All but one patient with a positive (DD ≥500 ng/ml) latex assay had DD levels higher than 500 ng/ml with the ELISA assay. Accordingly, ELISA technique could be restricted to patients with a negative result in latex assay. This two-step approach would have spared about 50% of ELISA in our cohort. In conclusion, our data indicate that a latex test can be used as a first diagnostic step to rule out pulmonary embolism provided a negative result is confirmed by ELISA and the performance of the latex assay used has been assessed properly.

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.

scholarly journals Epidemiological situation of bovine brucellosis in the State of Paraiba, Brazil

2016 ◽  
Vol 37 (5Supl2) ◽  
pp. 3403 ◽  
Author(s):  
Inácio José Clementino ◽  
Ricardo Augusto Dias ◽  
Marcos Amaku ◽  
Fernando Ferreira ◽  
Evelise Oliveira Telles ◽  
...  
Keyword(s):  
Risk Factor ◽  
Odds Ratio ◽  
The State ◽  
Prevalence Rates ◽  
Risk Of Infection ◽  
Bovine Brucellosis ◽  
Serum Samples ◽  
The Rose ◽  
Positive Results ◽  
Epidemiological Situation

This study was performed to characterize the epidemiological status of brucellosis in the State of Paraíba, Brazil. The State was divided into three regions. Herds were randomly sampled in each region and a pre-established number of animals were sampled in each of these herds. A total of 3,489 serum samples from 674 herds were collected. In each herd, an epidemiological questionnaire was conducted. This questionnaire focused on herd traits, as well as husbandry and sanitary practices that could be associated with the risk of infection. The serum samples were screened for antibodies against Brucella spp. by the Rose-Bengal Test (RBT), and all positive sera were confirmed by the 2-mercaptoethanol test (2-ME). The herd was considered positive if at least one animal had positive results for both the RBT and the 2-ME test. The prevalence rates of infected herds and animals in the State were 4.6% [3.2-6.5%] and 2.5% [1.1-3.9%], respectively. The prevalence rates of infected herds and animals in the regions were, respectively: region 1, 3.2% [1.5-6.6%] and 1.7% [0.5-5.7%]; region 2, 2.2% [0.9-5.2%] and 0.7% [0.3-1.7%]; and region 3, 7.9% [5.0-12.2%] and 3.2% [1.6-6.3%]. The risk factor (odds ratio, OR) associated with the presence of the infection was Zebuine as the predominant breed (OR=12.30 [1.32-114.64]).

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.

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