Detection of antibodies to caprine arthritis-encephalitis virus by protein G enzyme-linked immunosorbent assay and immunoblotting.

1989 ◽  
Vol 27 (3) ◽  
pp. 580-582 ◽  
Author(s):  
R Zanoni ◽  
A Krieg ◽  
E Peterhans
Keyword(s):  
Immunosorbent Assay ◽  
Encephalitis Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein G ◽  
Caprine Arthritis Encephalitis Virus

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 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 Investigation of caprine arthritis-encephalitis virus in Sudan using competitive enzyme-linked immunosorbent assay

2013 ◽  
Vol 6 (8) ◽  
pp. 558 ◽  
Author(s):  
Abdelghafar Elfahal ◽  
Mohammed Hussien ◽  
Khalid Enan ◽  
Khalid Taha ◽  
Diaeldin Salih ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Encephalitis Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Caprine Arthritis Encephalitis Virus

scholarly journals Evaluation of a Kinetic Enzyme-Linked Immunosorbent Assay for Detection of Caprine Arthritis-Encephalitis Virus-Specific Antibodies

1994 ◽  
Vol 6 (1) ◽  
pp. 30-33 ◽  
Author(s):  
J. Vander Schalie ◽  
D. S. Bradway ◽  
T. E. Besser ◽  
J. F. Evermann
Keyword(s):  
Immunosorbent Assay ◽  
Sodium Dodecyl ◽  
Encephalitis Virus ◽  
Cutoff Point ◽  
Enzyme Linked Immunosorbent Assay ◽  
Good Sensitivity ◽  
Diagnostic Laboratory ◽  
Caprine Arthritis Encephalitis Virus ◽  
Large Numbers ◽  
Turn Around Time

A kinetic indirect enzyme-linked immunosorbent assay (k-ELISA) was evaluated for detection of antibody to caprine arthritis-encephalitis virus (CAEV), using sodium dodecyl sulfate-treated CAEV-63 as antigen. Two hundred fifteen caprine sera submitted to the diagnostic laboratory were tested for CAEV antibody by the k-ELISA and by immunoprecipitation of [35S]-methionine-labeled CAEV. A k-ELISA positive cutoff point of 80 yielded a sensitivity of 94.4% and a specificity of 100%, as compared with immunoprecipitation. A k-ELISA cutoff point of 50 resulted in a sensitivity of 100%, with 95.6% specificity. When sera with k-ELISA scores between 50 and 80 were considered suspect, testing of 1,001 diagnostic sera resulted in < 1.5% suspect reactions. Using the 80 cutoff point, the CAEV k-ELISA had good sensitivity and specificity, with the added advantages of quick turn-around time, few suspect reactions, and adaptability to large numbers of samples.

scholarly journals Evaluation of a Caprine Arthritis-Encephalitis Virus/Maedi-Visna Virus Indirect Enzyme-Linked Immunosorbent Assay in the Serological Diagnosis of Ovine Progressive Pneumonia Virus in U.S. Sheep

2009 ◽  
Vol 17 (2) ◽  
pp. 307-310 ◽  
Author(s):  
Lynn M. Herrmann-Hoesing ◽  
Liam E. Broughton-Neiswanger ◽  
Kimberly C. Gouine ◽  
Stephen N. White ◽  
Michele R. Mousel ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Encephalitis Virus ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Caprine Arthritis Encephalitis Virus ◽  
Visna Virus ◽  
Maedi Visna Virus ◽  
Ovine Progressive Pneumonia

ABSTRACT A caprine arthritis-encephalitis virus (CAEV)/maedi-visna virus (MVV) indirect enzyme-linked immunosorbent assay (iELISA) was validated with samples from U.S. sheep and by the use of radioimmunoprecipitation as the standard for comparison. The sensitivity and the specificity were 86.0% (±5.8%) and 95.9% (±2.9%), respectively. The iELISA format and phylogenetic differences based on the MVV gag sequence contribute to the reduced sensitivity.

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.

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