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 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 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 Evaluation of a Blocking ELISA Using a Urease Conjugate for the Detection of Antibodies to Pseudorabies Virus

2000 ◽  
Vol 12 (3) ◽  
pp. 266-268 ◽  
Author(s):  
Maria Gabriela Echeverría ◽  
Edgardo Omar Nosetto ◽  
Maria Elisa Etcheverrigaray
Keyword(s):  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Pseudorabies Virus ◽  
Visual Assessment ◽  
Field Conditions ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Blocking Elisa ◽  
Elisa Format

A blocking enzyme-linked immunosorbent assay (ELISA) using a urease conjugate (U-B-ELISA) was evaluated for screening sera for antibodies to pseudorabies virus under field conditions. A total of 764 serum samples were analyzed by U-B-ELISA. Of these, 264 were evaluated by both virus neutralization and U-B-ELISA, and the results were compared. U-B-ELISA showed 98.5% and 98.9% sensitivity and specificity, respectively. This test combines the sensitivity and specificity of the blocking ELISA format while allowing visual assessment of results.

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 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.

Multicolor enzyme-linked immunoassay method for visual readout of carbendazim

2021 ◽  
Author(s):  
Haoran Liu ◽  
Yiwen Wang ◽  
Ruijie Fu ◽  
Jing Zhou ◽  
Yanlin Liu ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity

Enzyme-linked immunosorbent assay (ELISA) with high specificity and sensitivity is one of the most popular techniques for detecting carbendazim (CBD), a commonly used benzimidazole fungicide in agriculture. However, the traditional...

scholarly journals Prevalence of feline coronavirus antibodies in cats in Bursa province, Turkey, by an enzyme-linked immunosorbent assay

2009 ◽  
Vol 11 (10) ◽  
pp. 881-884 ◽  
Author(s):  
Annamaria Pratelli ◽  
Kadir Yesilbag ◽  
Marcello Siniscalchi ◽  
Ebru Yalçm ◽  
Zeki Yilmaz
Keyword(s):  
Immunosorbent Assay ◽  
Predictive Value ◽  
Gold Standard ◽  
Population Based ◽  
Standard Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
High Association ◽  
Gold Standard Test ◽  
Feline Coronavirus

Feline sera from Bursa province (Turkey) were assayed for coronavirus antibody using an enzyme-linked immunosorbent assay (ELISA). The study was performed on 100 sera collected from cats belonging to catteries or community shelters and to households. The serum samples were initially tested with the virus neutralisation (VN) test and the results were then compared with the ELISA. The VN yielded 79 negative and 21 positive sera but the ELISA confirmed only 74 as negative. The ELISA-negative sera were also found to be free of feline coronoviruses-specific antibodies by Western blotting. Using the VN as the gold standard test, ELISA had a sensitivity of 100% and a specificity of 93.6%, with an overall agreement of 95%. The Kappa (κ) test indicated high association between the two tests (κ=0.86, 95% confidence interval (CI) 0.743–0.980). The positive predictive value (PPV) was 0.8, and the negative predictive value (NPV) was 0.93. The prevalence of FCoV II antibodies in the sampled population based on the gold standard was 62% (95% CI 0.44–0.77) among multi-cat environments, and 4% (95% CI 0.01–0.11) among single cat households.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Antibody to Epizootic Hemorrhagic Disease of Deer Virus

2000 ◽  
Vol 12 (2) ◽  
pp. 142-145 ◽  
Author(s):  
James O. Mecham ◽  
Michael M. Jochim
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hemorrhagic Disease ◽  
Structural Protein ◽  
Serum Samples ◽  
Epizootic Hemorrhagic Disease ◽  
Diagnostic Potential ◽  
The Us ◽  
Serotype 1 ◽  
Infected Cells

An enzyme-linked immunosorbent assay has been developed to detect antibodies to epizootic hemorrhagic disease of deer virus (EHDV). The assay incorporates a monoclonal antibody to EHDV serotype 2 (EHDV-2) that demonstrates specificity for the viral structural protein, VP7. The assay was evaluated with sequential sera collected from cattle experimentally infected with EHDV serotype 1 (EHDV-1) and EHDV-2, as well as the four serotypes of bluetongue virus (BTV), BTV-10, BTV-11, BTV-13, and BTV-17, that currently circulate in the US. A competitive and a blocking format as well as the use of antigen produced from both EHDV-1-and EHDV-2-infected cells were evaluated. The assay was able to detect specific antibody as early as 7 days after infection and could differentiate animals experimentally infected with EHDV from those experimentally infected with BTV. The diagnostic potential of this assay was demonstrated with field-collected serum samples from cattle, deer, and buffalo.

Sign in / Sign up

Export Citation Format

Share Document