scholarly journals Competitive-Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Serum Antibodies to Caprine Arthritis-Encephalitis Virus: Diagnostic Tool for Successful Eradication

2003 ◽  
Vol 10 (2) ◽  
pp. 267-271 ◽  
Author(s):  
Lynn M. Herrmann ◽  
William P. Cheevers ◽  
Travis C. McGuire ◽  
D. Scott Adams ◽  
Melinda M. Hutton ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
False Negative ◽  
The United States ◽  
Encephalitis Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dairy Goats ◽  
Serum Antibodies ◽  
Serum Samples ◽  
Caprine Arthritis Encephalitis Virus

ABSTRACT A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was evaluated for the detection of serum antibodies to the surface envelope (SU) of caprine arthritis-encephalitis virus (CAEV) in goats. This assay utilized 96-well microtiter plates containing CAEV-63 SU captured by monoclonal antibody (MAb) F7-299 and measured the competitive displacement of horseradish peroxidase-conjugated MAb GPB 74A binding by undiluted goat sera (F. Özyörük, W. P. Cheevers, G. A. Hullinger, T. C. McGuire, M. Hutton, and D. P. Knowles, Clin. Diagn. Lab. Immunol. 8:44-51, 2001). Two hundred serum samples from goats in the United States were used to determine the sensitivity and specificity of cELISA based on the immunoprecipitation (IP) of [35S]methionine-labeled viral antigens as a standard of comparison. A positive cELISA was defined as >33.2% inhibition of MAb 74A binding based on 2 standard deviations above the mean percent inhibition of 140 IP-negative serum samples. At this cutoff value, there were 0 of 60 false-negative sera (100% sensitivity) and 5 of 140 false-positive sera (96.4% specificity). Additional studies utilized IP-monitored cELISA to establish a CAEV-free herd of 1,640 dairy goats.

scholarly journals Detection of Serum Antibodies to Ovine Progressive Pneumonia Virus in Sheep by Using a Caprine Arthritis-Encephalitis Virus Competitive-Inhibition Enzyme-Linked Immunosorbent Assay

2003 ◽  
Vol 10 (5) ◽  
pp. 862-865 ◽  
Author(s):  
Lynn M. Herrmann ◽  
William P. Cheevers ◽  
Katherine L. Marshall ◽  
Travis C. McGuire ◽  
Melinda M. Hutton ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
False Negative ◽  
High Sensitivity ◽  
The United States ◽  
Encephalitis Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Caprine Arthritis Encephalitis Virus ◽  
Ovine Progressive Pneumonia

ABSTRACT A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) for detection of antibodies to the surface envelope (SU) of caprine arthritis-encephalitis virus (CAEV) was recently reported (L. M. Herrmann, W. P. Cheevers, T. C. McGuire, D. Scott Adams, M. M. Hutton, W. G. Gavin, and D. P. Knowles, Clin. Diagn. Lab. Immunol. 10:267-271, 2003). The cELISA utilizes CAEV-63 SU captured on microtiter plates using the monoclonal antibody (MAb) F7-299 and measures competitive displacement of binding of the anti-CAEV MAb GPB 74A by goat serum. The present study evaluated the CAEV cELISA for detection of antibodies to ovine progressive pneumonia virus (OPPV) in sheep. Three hundred thirty-two sera were randomly selected from 21,373 sheep sera collected throughout the United States to determine the sensitivity and specificity of cELISA and agar gel immunodiffusion (AGID) based on immunoprecipitation (IP) of [35S]methionine-labeled OPPV antigens as a standard of comparison. A positive cELISA test was defined as >20.9 percent inhibition (% I) of MAb 74A binding based on two standard deviations above the mean % I of 191 IP-negative sheep sera. At this cutoff, there were 2 of 141 false-negative sera (98.6% sensitivity) and 6 of 191 false-positive sera (96.9% specificity). Sensitivity and specificity values for IP-monitored AGID were comparable to those for cELISA for 314 of 332 sera with unambiguous AGID results. Concordant results by cELISA and IP resolved 16 of the 18 sera that were indeterminate by AGID. Additional studies evaluated cELISA by using 539 sera from a single OPPV-positive flock. Based on IP of 36 of these sera, there was one false-negative by cELISA among 21 IP-positive sera (95.5% sensitivity) and 0 of 15 false-positives (100% specificity). We conclude that the CAEV cELISA can be applied to detection of OPPV antibodies in sheep with high sensitivity and specificity.

scholarly journals Monoclonal Antibodies to Conformational Epitopes of the Surface Glycoprotein of Caprine Arthritis-Encephalitis Virus: Potential Application to Competitive-Inhibition Enzyme-Linked Immunosorbent Assay for Detecting Antibodies in Goat Sera

2001 ◽  
Vol 8 (1) ◽  
pp. 44-51 ◽  
Author(s):  
Fuat Özyörük ◽  
William P. Cheevers ◽  
Gordon A. Hullinger ◽  
Travis C. McGuire ◽  
Melinda Hutton ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
Encephalitis Virus ◽  
Surface Glycoprotein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Western Blots ◽  
Caprine Arthritis Encephalitis Virus ◽  
Potential Applicability ◽  
Surface Envelope

ABSTRACT Four immunoglobulin G1 monoclonal antibodies (MAbs) to the gp135 surface envelope glycoprotein (SU) of the 79–63 isolate of caprine arthritis-encephalitis virus (CAEV), referred to as CAEV-63, were characterized and evaluated for their ability to compete with antibody from CAEV-infected goats. Three murine MAbs (MAbs GPB16A, 29A, and 74A) and one caprine MAb (MAb F7-299) were examined. All MAbs reacted in nitrocellulose dot blots with native CAEV-63 SU purified by MAb F7-299 affinity chromatography, whereas none reacted with denatured and reduced SU. All MAbs reacted in Western blots with purified CAEV-63 SU or the SU component of whole-virus lysate following denaturation in the absence of reducing agent, indicating that intramolecular disulfide bonding was essential for epitope integrity. Peptide-N-glycosidase F digestion of SU abolished the reactivities of MAbs 74A and F7-299, whereas treatment of SU withN-acetylneuraminate glycohydrolase (sialidase A) under nonreducing conditions enhanced the reactivities of all MAbs as well as polyclonal goat sera. MAbs 29A and F7-299 were cross-reactive with the SU of an independent strain of CAEV (CAEV-Co). By enzyme-linked immunosorbent assay (ELISA), the reactivities of horseradish peroxidase (HRP)-conjugated MAbs 16A and 29A with homologous CAEV-63 SU were <10% of that of HRP-conjugated MAb 74A. The reactivity of HRP-conjugated MAb 74A was blocked by sera from goats immunized with CAEV-63 SU or infected with CAEV-63. The reactivity of MAb 74A was also blocked by sera from goats infected with a CAEV-Co molecular clone, although MAb 74A did not react with CAEV-Co SU in Western blots. Thus, goats infected with either CAEV-63 or CAEV-Co make antibodies that inhibit binding of MAb 74A to CAEV-63 SU. A competitive-inhibition ELISA based on displacement of MAb 74A reactivity has potential applicability for the serologic diagnosis of CAEV infection.

scholarly journals  Toxoplasma gondii and Neospora caninum antibodies in goats in the Czech Republic

2012 ◽  
Vol 57 (No. 3) ◽  
pp. 111-114 ◽  
Author(s):  
E. Bartova ◽  
K. Sedlak
Keyword(s):  
Czech Republic ◽  
Toxoplasma Gondii ◽  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
Neospora Caninum ◽  
Fluorescent Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Antibodies ◽  
Serum Samples ◽  
The Czech Republic

Toxoplasma gondii is zoonotic protozoan parasite that causes infections in many vertebrate species. The present study determined the seroprevalence of T. gondii and N. caninum in goats from the Czech Republic. Serum samples were collected from 251 healthy adult goats in the Czech Republic during the years 2006 to 2009. Sera samples were tested for serum antibodies to Toxoplasma gondii by an enzyme-linked immunosorbent assay with cut off equal to or higher than 50% S/P. The same samples were tested for serum antibodies to Neospora caninum by a competitive-inhibition enzyme-linked immunosorbent assay with cut off equal to or higher than 30% inhibition; positive sera were confirmed by an indirect fluorescent antibody test with cut-off titre equal to or higher than 40. Sera positive in both tests were marked as positive. In total, 166 (66%) and 15 (6%) goat sera reacted positively for T. gondii and N. caninum antibodies, respectively. All sera positive for N. caninum antibodies were simultaneously positive for T. gondii antibodies. This is the&nbsp;first detection of N.&nbsp;caninum antibodies in goats in the Czech Republic. Our findings indicate that goats in the Czech Republic are frequently exposed to T.&nbsp;gondii, but less frequently to N. caninum. &nbsp;

scholarly journals Seroconversion and seroreactivity patterns of dairy goats naturally exposed to caprine arthritis-encephalitis virus in Brazil

2002 ◽  
Vol 32 (4) ◽  
pp. 603-607 ◽  
Author(s):  
Roberto Soares Castro ◽  
Rômulo Cerqueira Leite ◽  
Edisio Oliveira de Azevedo ◽  
Maurício Resende ◽  
Aurora Maria Guimarães Gouveia
Keyword(s):  
Seroconversion Rate ◽  
Individual Variability ◽  
Encephalitis Virus ◽  
Lactation Period ◽  
Dairy Goats ◽  
Serum Samples ◽  
Caprine Arthritis Encephalitis Virus ◽  
Standard Curve

A labelled avidin-biotin ELISA (lab-ELISA) using repeated serum samples of goats showed a progressive seroconversion with higher seroconversion rate at the period going from the beginning of the breeding up to the last half of lactation (35.0%), compared to that recorded at the beginning of breeding (17.8%)(p<0.05). Furthermore, the seroreactivity pattern, evaluated by a lab-ELISA standard-curve with serum samples collected at 30-40 days intervals during 12 months, was caracterized by high individual variability. No seroreversion was observed and there were higher titers in the group of animals which delivered kids and established a lactation period (n=6; mean titre=913.4 units) compared to the group of goats that failed to conceived (n=4; mean titre=261.2 units) (p<0.01).

scholarly journals Initial multi-target approach shows importance of improved caprine arthritis-encephalitis virus control program in Russia for hobbyist goat farms

2021 ◽  
pp. 1718-1726
Author(s):  
Eduard A. Shuralev ◽  
Nail I. Khammadov ◽  
Konstantin A. Osyanin ◽  
Inna A. Elizarova ◽  
Gaysha R. Salmanova ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Control Program ◽  
Encephalitis Virus ◽  
Polymerase Chain Reaction Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Caprine Arthritis Encephalitis Virus ◽  
The Third ◽  
Molecular Tests

Background and Aim: Several reports described the detection of specific caprine arthritis-encephalitis virus (CAEV) antibodies in Russian goat populations, which indicates the circulation of CAEV in Russian goat farms. The aim of this study was to use a multi-target approach to testing with both serological tests and an in-house real-time (RT) molecular test to investigate the prevalence of CAEV in goats from three hobbyist farms in the Republic of Tatarstan, Russia. Materials and Methods: We applied a multi-target approach to testing with both enzyme-linked immunosorbent assay (ELISA) and an in-house RT polymerase chain reaction test to investigate the prevalence of CAEV in goats. Animals from the three hobbyist farms were used in this study. The animals from two farms (n=13 for F1 and n=8 for F2) had clinical signs of arthritis and mastitis. In the third farm (n=15 for F3), all goats were home-bred and had no contact with imported animals. Results: CAEV antibodies (ELISA targets TM env and gag genes) were detected in serum samples from two farms (F1 and F2), indicating seroprevalence of 87.50-92.31%. Specific CAEV antibodies were also detected in milk samples. CAEV proviral DNA was detected in 53.85-62.50%. The results from all tests performed in the third farm (F3) were negative, indicating that all tests were 100% specific. Conclusion: The results showed that CAEV is circulating and present in small hobbyist goat farms in Russia. Serological and molecular tests could be important for programs to control and eradicate CAEV in Russia for hobbyist goat farms.

scholarly journals Evaluation of a Diagnostic Algorithm Using Immunoglobulin M Enzyme-Linked Immunosorbent Assay To Differentiate Human West Nile Virus and St. Louis Encephalitis Virus Infections during the 2002 West Nile Virus Epidemic in the United States

2004 ◽  
Vol 11 (6) ◽  
pp. 1130-1133 ◽  
Author(s):  
Denise A. Martin ◽  
Amanda Noga ◽  
Olga Kosoy ◽  
Alison J. Johnson ◽  
Lyle R. Petersen ◽  
...  
Keyword(s):  
United States ◽  
West Nile Virus ◽  
Immunosorbent Assay ◽  
Diagnostic Algorithm ◽  
The United States ◽  
Encephalitis Virus ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
West Nile

ABSTRACT A diagnostic algorithm was developed to differentiate between human infections of West Nile virus (WNV) and St. Louis encephalitis virus (SLEV) using positive-to-negative (P/N) ratios derived from the immunoglobulin M capture enzyme-linked immunosorbent assay (MAC-ELISA). To validate this algorithm, we tested 1,418 serum and cerebrospinal fluid (CSF) samples from confirmed WNV and SLEV infections collected during the WNV epidemic of 2002 in the United States. WNV P/N-to-SLEV P/N ratios (W/S ratios) were calculated and used to identify the infecting virus. These results were compared to results from the plaque reduction neutralization test (PRNT), which is currently the standard assay used to discriminate between closely related flavivirus infections. If the W/S ratio was ≥1, the predictive value positive (PNP) for WNV was 97.8%, where 95% of flavivirus cases were due to WNV infection and only 3.7% of specimens would require PRNT to differentiate WNV from SLEV infection. Use of the W/S ratio as part of the testing algorithm to interpret MAC-ELISA results generates reportable probable cases quickly, alleviating the need for PRNT in most instances.

scholarly journals Detection of Anaplasma antibodies in wildlife and domestic species in wildlife-livestock interface areas of Kenya by major surface protein 5 competitive inhibition enzyme-linked immunosorbent assay

2008 ◽  
Vol 75 (3) ◽  
Author(s):  
J.J.N. Ngeranwa ◽  
S.P. Shompole ◽  
E.H. Venter ◽  
A. Wambugu ◽  
J.E. Crafford ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
Surface Membrane ◽  
Clinical Signs ◽  
Surface Protein ◽  
Domestic Animal ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Domestic Species ◽  
Major Surface

The seroprevalence of Anaplasma antibodies in wildlife (eland, blue wildebeest, kongoni, impala, Thomson's gazelle, Grant's gazelle, giraffe and plains zebra) and domestic animal (cattle, sheep and goat) populations was studied in wildlife / livestock interface areas of Kenya. Serum samples were analyzed by competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA), using a recombinant antigen (MSP-5) from Anaplasma marginale surface membrane. A monoclonal antibody, FC-16, was used as the primary antibody, while anti-mouse conjugated to horseradish peroxidase was used as the secondary antibody. The results indicate a high seroprevalence in both wildlife and livestock populations, in contrast to earlier reports from Kenya, which indicated a low seroprevalence. The differences are attributed to the accurate analytical method used (CI-ELISA), as compared with agglutination techniques, clinical signs and microscopy employed by the earlier workers.

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