scholarly journals Lipopolysaccharide-Based Enzyme-Linked Immunosorbent Assay for Experimental Use in Detection of Antibodies to Lawsonia intracellularis in Pigs

2005 ◽  
Vol 12 (6) ◽  
pp. 693-699 ◽  
Author(s):  
J. J. Kroll ◽  
M. A. Eichmeyer ◽  
M. L. Schaeffer ◽  
S. McOrist ◽  
D. L. Harris ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Experimental Studies ◽  
Antibody Test ◽  
Controlled Study ◽  
Future Research ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lawsonia Intracellularis ◽  
Serum Samples ◽  
Specificity And Sensitivity

ABSTRACT An enzyme-linked immunosorbent assay (ELISA) for Lawsonia intracellularis was developed and compared with a whole-cell antigen-based immunofluorescence antibody test (IFAT). The antigen-containing lipopolysaccharide (LPS) was derived from Percoll gradient purified cultures of L. intracellularis by using a modification of the Westphal hot phenol procedure. The antigen was bound directly to polystyrene 96-well microtiter plates, and the assay was performed in an indirect ELISA format. Specificity and sensitivity values based on 80 known positive and 80 known negative serum samples from controlled experimental trials were 93.7% and 88.7%, respectively. Serological results from a controlled L. intracellularis challenge exposure study confirmed the high specificity and sensitivity of this assay (100% and 99.5%, respectively). Comparisons between the LPS ELISA and the IFAT in detecting anti-Lawsonia antibodies in this controlled study revealed significantly more LPS ELISA-positive pigs than IFAT-positive pigs on days 21, 28, 35, and 42 (P = 0.003, 0.030, 0.002, and 0.006, respectively). This indirect ELISA (LPS ELISA) test is an improved method of detecting antibodies in pigs soon after exposure to L. intracellularis, regardless of isolate type (vaccine or wild type) in experimental studies. The LPS ELISA may be used as a tool to support future research trials on vaccine efficacy and to further understand the immune response induced by L. intracellularis.

scholarly journals An enzyme-linked immunosorbent assay for the detection of IgG antibodies against Babesia equi in horses

2004 ◽  
Vol 34 (5) ◽  
pp. 1525-1529 ◽  
Author(s):  
Cristiane Divan Baldani ◽  
Rosangela Zacarias Machado ◽  
Paulo de Tarso Landgraf Botteon ◽  
Felipe Santoro Takakura ◽  
Carlos Luiz Massard
Keyword(s):  
Immunosorbent Assay ◽  
Serological Test ◽  
Epidemiological Studies ◽  
Sao Paulo ◽  
Enzyme Linked Immunosorbent Assay ◽  
São Paulo ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Babesia Equi ◽  
Specificity And Sensitivity

A crude antigenic preparation of Babesia equi was used to develop and establish the suitability of an enzyme-linked immunosorbent assay (ELISA) for the detection of parasite carriers. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 90 serum samples were taken from horses from the Northeast region of São Paulo State and examined for diagnosis of equine B. equi infection by ELISA. Approximately 75% (n=67) of all the horses tested were found serologically positive for B. equi. These results suggest that the ELISA described may prove to be an appropriate serological test for epidemiological studies on B. equi infections in the field and that equine piroplasmosis is a cause for serious concern in the State of São Paulo, Brazil.

scholarly journals Development of Rapid Immunochromatographic Test with Recombinant NcSAG1 for Detection of Antibodies to Neospora caninum in Cattle

2005 ◽  
Vol 12 (7) ◽  
pp. 885-887 ◽  
Author(s):  
Min Liao ◽  
Shoufa Zhang ◽  
Xuenan Xuan ◽  
Guohong Zhang ◽  
Xiaohong Huang ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Reliable Method ◽  
Neospora Caninum ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunochromatographic Test ◽  
Serum Samples ◽  
Immunofluorescent Antibody Test ◽  
Good Agreement

ABSTRACT An immunochromatographic test (ICT) with recombinant surface antigen 1 of Neospora caninum (NcSAG1) was developed for the rapid detection of antibodies to N. caninum in cattle. The ICT was used to clearly discriminate between immunofluorescent-antibody test (IFAT)-positive bovine sera and IFAT-negative bovine sera. Serum samples collected from cattle in Yanbian, China, were examined by the ICT. Of the 96 serum samples, 23 (24.0%) were positive by the ICT, and 19 (19.8%) samples were positive by a previously developed enzyme-linked immunosorbent assay (ELISA). Eighteen of 19 ELISA-positive samples were positive according to the ICT. A good agreement was found between the results of the ICT and the ELISA. The results presented here suggest that the ICT with recombinant truncated NcSAG1 fused to glutathione S-transferase is a useful and reliable method for the detection of antibodies to N. caninum in cattle.

Development and preliminary application of gE enzyme-linked immunosorbent assay for detection of the antibody to gE protein ofPseudorabies virusin pigs

2005 ◽  
Vol 2 (2) ◽  
pp. 79-83 ◽  
Author(s):  
Tang Yong ◽  
Chen Huan-Chun ◽  
Qin Ya-Li ◽  
He Qi-Gai ◽  
Jin Mei-Lli ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Pseudorabies Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Specificity ◽  
Serum Samples ◽  
Agreement Rate ◽  
Glycoprotein E ◽  
Recombinant Glycoprotein ◽  
Specificity And Sensitivity ◽  
Preliminary Application

AbstractTo differentiate pigs infected withPseudorabies virus(PrV) from pigs vaccinated with gE-PrV, a glycoprotein E enzyme-linked immunosorbent assay (gE-ELISA) based on recombinant glycoprotein E (gE) (which was expressed byEscherichia coli, purified, denatured and renatured) was developed. By testing 115 serum samples, the diagnostic specificity and sensitivity of the developed gE-ELISA were evaluated to be 94.5% and 96.7%, respectively. Five serum samples were tested with plates from five lots, and the results had a coefficient of variation of less than 10%, showing good reproducibility of gE-ELISA. This gE-ELISA was compared with a commercial blocking ELISA by testing 356 serum samples. The agreement rate of the two assays was 92.13% (328/356). These results suggested that the gE-ELISA developed in our laboratory could be used in differentiating PrV-infected and gE-PrV-vaccinated pigs.

scholarly journals A Reverse-Sandwich Enzyme-Linked Immunosorbent Assay for Verocytotoxin 1 and 2 Antibodies in Human and Bovine Sera

1999 ◽  
Vol 6 (5) ◽  
pp. 701-704 ◽  
Author(s):  
H. Miyazawa ◽  
H. Bannai ◽  
T. Yanase ◽  
C. Morita ◽  
S. Satoh ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Immunosorbent Assay ◽  
Animal Species ◽  
Indirect Elisa ◽  
Dairy Farm ◽  
Normal Adult ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity ◽  
Seroepidemiologic Studies ◽  
Adult Humans

ABSTRACT A reverse-sandwich enzyme-linked immunosorbent assay (ELISA), in which an antibody is sandwiched by antigens, was established for the titration of antibodies to verocytotoxins (VT) in human and animal sera. This assay has two advantages over a conventional indirect ELISA: (i) higher specificity and sensitivity and (ii) the ability to comparably titrate antibodies from different species. The VT1 (Shiga-like toxin 1) antibody-positive rates were 5% in 202 normal adult humans and 99% in 93 normal cattle at a dairy farm. This ELISA is most suitable for seroepidemiologic studies of infections with VT-producing Escherichia coli in humans and various animal species.

scholarly journals Rapid Competitive Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Peste des Petits Ruminants Virus

2005 ◽  
Vol 12 (4) ◽  
pp. 542-547 ◽  
Author(s):  
Kang-Seuk Choi ◽  
Jin-Ju Nah ◽  
Young-Joon Ko ◽  
Shien-Young Kang ◽  
Nam-In Jo
Keyword(s):  
Immunosorbent Assay ◽  
Small Ruminants ◽  
Viral Disease ◽  
Turnaround Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peste Des Petits Ruminants ◽  
Rinderpest Virus ◽  
Serum Samples ◽  
Specificity And Sensitivity ◽  
Relative Specificity

ABSTRACT Peste des petits ruminants (PPR) is a contagious viral disease of small ruminants that is of economic importance in Africa, the Middle East, and Asia. We developed a rapid competitive enzyme-linked immunosorbent assay (rapid c-ELISA) for the diagnosis and surveillance of PPR. This assay detects PPR virus (PPRV) antibodies in serum samples by quantifying the amount of monoclonal antibody (MAb) P-3H12 after 30 min of incubation of a serum-MAb conjugate mixture on plates coated with a PPRV recombinant nucleocapsid protein (rPPRV-N). We tested 249 PPRV-positive serum samples and 733 PPRV-negative serum samples from field ruminants. The threshold of percent inhibition (PI) was determined to be <50 on the basis of the mean PI plus 3 standard deviations for sera from PPRV-negative ruminants. The relative specificity and sensitivity of the rapid c-ELISA were 98.5% (722 of 733 serum samples) and 93.4% (234 of 249 serum samples), respectively. The rapid c-ELISA sensitively detected PPRV antibodies in hyperimmune sera (virus neutralization test [VNT] titer, >512), even at dilutions ≥512 in normal goat serum, and as early as 6 to 13 days postinfection from 12 goats, each of which was infected with one of the four PPRV lineages. Hyperimmune sera from animals experimentally vaccinated with rinderpest virus gave positive results by the rapid c-ELISA when the rinderpest virus VNT titers were >512, although the rapid c-ELISA titers were very low (2 to 16). However, the rapid c-ELISA was negative when the rinderpest virus VNT titer was ≤128. The rapid c-ELISA developed in the present work provides a short turnaround time and could be a useful tool for the diagnosis of PPR and screening for PPRV in the field.

scholarly journals An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against Leishmania chagasi in dogs

2009 ◽  
Vol 29 (2) ◽  
pp. 120-124 ◽  
Author(s):  
Débora Carvalho ◽  
Trícia M.F.S. Oliveira ◽  
Cristiane D. Baldani ◽  
Rosangela Z. Machado
Keyword(s):  
Visceral Leishmaniasis ◽  
Immunosorbent Assay ◽  
Fluorescent Antibody ◽  
Domestic Dog ◽  
Enzyme Linked Immunosorbent Assay ◽  
Leishmania Chagasi ◽  
Serum Samples ◽  
Igm Antibodies ◽  
Specificity And Sensitivity ◽  
Belo Horizonte

Visceral leishmaniasis is an emergent zoonosis with an increasing number of new cases in Brazil where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. An enzyme-linked immunosorbent assay (ELISA), based upon the use of a total soluble antigenic preparation of L. chagasi, was adapted for the detection of IgM antibodies in the serum of infected dogs. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 110 serum samples were taken from dogs in Belo Horizonte, Minas Gerais, Brazil, and examined for anti-L. chagasi IgM antibody by ELISA and indirect fluorescent antibody test (IFAT). About 25% (n=27) of all the dogs tested were found serologically positive for L. chagasi by IFAT, while 89.09% (n=98) were seropositive by ELISA. The results obtained by ELISA and IFAT were significantly different (P<0.01). The combined use of ELISA and IFAT is recommended in order to enable veterinary services to more efficiently detect canine visceral leishmaniasis.

scholarly journals Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

2020 ◽  
Author(s):  
Chanakan Areewong ◽  
Amarin Rittipornlertrak ◽  
Boondarika Nambooppha ◽  
Itsarapan Fhaikrue ◽  
Tawatchai Singhla ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Hemagglutination Inhibition ◽  
Indirect Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Panthera Tigris ◽  
Serum Samples ◽  
Feline Panleukopenia Virus ◽  
Elisa Testing ◽  
Antibody Levels

Abstract Background: Feline panleukopenia virus (FPV) is an etiologic pathogen of feline panleukopenia that infects all members of Felidae including tigers (Panthera tigris). Vaccination against FPV among wild felid species has long been practiced in zoos worldwide. However, few studies have assessed tiger immune response post-vaccination due to the absence of a serological diagnostic tool. To address these limitations, this study aimed to develop an in-house indirect enzyme-linked immunosorbent assay (ELISA) for the monitoring of tiger antibody levels against the feline panleukopenia vaccine by employing the synthesized subunit capsid protein VP2. An in-house horseradish peroxidase (HRP) conjugated rabbit anti-tiger immunoglobulin G (IgG) polyclonal antibody (HRP-anti-tiger IgG) was produced in this study and employed in the assay. It was then compared to a commercial HRP-conjugated goat anti-cat IgG (HRP-anti-cat IgG). Sensitivity and specificity were evaluated using the Bayesian model with conditional dependence being assumed between both HRP-conjugated antibody-based ELISAs and hemagglutination-inhibition (HI) tests. Results: The posterior estimates for sensitivity and specificity of two indirect ELISA HRP-conjugated antibodies were higher than those of the HI test. The sensitivity and specificity of indirect ELISA for HRP-anti-tiger IgG and HRP-anti-cat IgG were 86.5%, 57.2% and 86.7%, 64.6%, respectively, while the results of the HI test were 79.1 and 54.1%. In applications, 89.6% (198/221) of tiger serum samples were determined to be seropositive by indirect ELISA testing. Conclusion: The results support evidence that an in-house indirect ELISA developed in this study could be used as a serological tool for the effective detection of tiger antibodies.

scholarly journals A Highly Sensitive and Subspecies-Specific Surface Antigen Enzyme- Linked Immunosorbent Assay for Diagnosis of Johne's Disease

2006 ◽  
Vol 13 (8) ◽  
pp. 837-844 ◽  
Author(s):  
Shigetoshi Eda ◽  
John P. Bannantine ◽  
W. R. Waters ◽  
Yasuyuki Mori ◽  
Robert H. Whitlock ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Surface Antigens ◽  
Johne's Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Specificity ◽  
Serum Samples ◽  
Low Level ◽  
Specificity And Sensitivity ◽  
Highly Sensitive ◽  
High Level

ABSTRACT Johne's disease (JD), or paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis, is one of the most widespread and economically important diseases of livestock and wild ruminants worldwide. Control of JD could be accomplished by diagnosis and good animal husbandry, but this is currently not feasible because commercially available diagnostic tests have low sensitivity levels and are incapable of diagnosing prepatent infections. In this study, a highly sensitive and subspecies-specific enzyme-linked immunosorbent assay was developed for the diagnosis of JD by using antigens extracted from the surface of M. avium subsp. paratuberculosis. Nine different chemicals and various intervals of agitation by vortex were evaluated for their ability to extract the surface antigens. Various quantities of surface antigens per well in a 96-well microtiter plate were also tested. The greatest differences in distinguishing between JD-positive and JD-negative serum samples by ethanol vortex enzyme-linked immunosorbent assay (EVELISA) were obtained with surface antigens dislodged from 50 μg/well of bacilli treated with 80% ethanol followed by a 30-second interval of agitation by vortex. The diagnostic specificity and sensitivity of the EVELISA were 97.4% and 100%, respectively. EVELISA plates that had been vacuum-sealed and then tested 7 weeks later (the longest interval tested) had diagnostic specificity and sensitivity rates of 96.9 and 100%, respectively. In a comparative study involving serum samples from 64 fecal culture-positive cattle, the EVELISA identified 96.6% of the low-level fecal shedders and 100% of the midlevel and high-level shedders, whereas the Biocor ELISA detected 13.7% of the low-level shedders, 25% of the mid-level shedders, and 96.2% of the high-level shedders. Thus, the EVELISA was substantially superior to the Biocor ELISA, especially in detecting low-level and midlevel shedders. The EVELISA may form the basis for a highly sensitive and subspecies-specific test for the diagnosis of JD.

scholarly journals Detection of tetanus toxoid antibodies in human sera in New Zealand by ELISA

1987 ◽  
Vol 98 (2) ◽  
pp. 199-202 ◽  
Author(s):  
R. C. H. Lau
Keyword(s):  
New Zealand ◽  
Human Serum ◽  
Immunosorbent Assay ◽  
Tetanus Toxoid ◽  
Indirect Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Age Group ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Human Sera

SUMMARYAn enzyme-linked immunosorbent assay (ELISA) incorporating the sensitive biotin-streptavidin system was developed to detect IgG antibodies to tetanus toxoid in human serum. Serum samples obtained from 557 normal persons aged 1–65 years from different areas in New Zealand were tested. The proportion of those immune ranged from 60–93% in males, and from 46–86% in females. In the 1–9 years age group 85% were immune. The indirect ELISA is suitable for serological surveys as it is simple to perform, economical and reproducible.

scholarly journals Development and Validation of A Competitive ELISA Based on Virus-Like Particles of Serotype Senecavirus A to Detect Serum Antibodies

2020 ◽  
Author(s):  
Manyuan Bai ◽  
Rui Wang ◽  
Shiqi Sun ◽  
Yun Zhang ◽  
Hu Dong ◽  
...  
Keyword(s):  
Expression System ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Virus Like Particles ◽  
Specificity And Sensitivity ◽  
Indirect Immunofluorescent Assay ◽  
Viral Particles ◽  
Test Kit ◽  
Development And Validation

Abstract Virus-like particles (VLPs) are high-priority antigens with highly ordered repetitive structures, which are similar to natural viral particles. We have developed a competitive enzyme-linked immunosorbent assay (cELISA) for detecting antibodies directed against Senecavirus A (SVA). Our assay utilizes SVA VLPs that were expressed and assembled in an Escherichia coli expression system as the coating antigens. VLPs have better safety and immunogenicity than intact viral particles or peptides. The VLP-based cELISA was used to test 342 serum samples collected from different pig farms, and the results showed that its specificity and sensitivity were 100% and 94%, respectively. The consistency rates of cELISA with the BIOSTONE AsurDx™ Senecavirus A (SVA) Antibody Test Kit and an indirect immunofluorescent assay were 90.0% and 94.2%, respectively. Therefore, this VLP-based cELISA can be effectively and reliably used for the detection and discrimination of SVA infection in serum samples.

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