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 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 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 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 Excretory-Secretory Antigens of Trypanosoma cruzi Are Potentially Useful for Serodiagnosis of Chronic Chagas' Disease

2001 ◽  
Vol 8 (5) ◽  
pp. 1024-1027 ◽  
Author(s):  
Mineo Nakazawa ◽  
Daniela S. Rosa ◽  
Valéria R. A. Pereira ◽  
Milena O. Moura ◽  
Veridiana C. Furtado ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chronic Chagas Disease ◽  
Secreted Antigens

ABSTRACT The reactivities of sera from chronic chagasic patients against the trypomastigote excreted-secreted antigens (TESA) of Trypanosoma cruzi strains with different biodemes were analyzed by TESA-blot and TESA–enzyme-linked immunosorbent assay (ELISA). Although both tests presented high sensitivity and specificity, TESA-ELISA is more appropriate for screening a larger number of samples.

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.

Screening Procedures for Clenbuterol Residue Determination in Raw Swine Livers Using Lateral-Flow Assay and Enzyme-Linked Immunosorbent Assay

2008 ◽  
Vol 71 (4) ◽  
pp. 865-869 ◽  
Author(s):  
WEI H. LAI ◽  
DANIEL Y. C. FUNG ◽  
YANG XU ◽  
YONG H. XIONG
Keyword(s):  
Immunosorbent Assay ◽  
False Positive Rate ◽  
False Negative ◽  
Screening Method ◽  
The United States ◽  
Lateral Flow ◽  
Enzyme Linked Immunosorbent Assay ◽  
Surveillance Program ◽  
The European Union ◽  
Lateral Flow Strip

Clenbuterol, which may cause symptoms of increased heart rate, muscular tremors, headache, nausea, and muscular cramps in patients, has been prohibited for consumption in many countries including the European Union, the United States, and China. A rapid lateral-flow strip assay was developed in our laboratory, and results obtained with this assay were compared with those obtained with a commercial enzyme-linked immunosorbent assay (ELISA) kit for the screening of clenbuterol in raw swine liver. A total of 128 swine livers were acquired from five local markets and prepared for analysis by the lateral-flow strip assay and ELISA. Analysis was completed in 10 min with the lateral-flow strip assay and in 90 min with the ELISA. In parallel with the ELISA, the rapid detection strip produced no false-negative results but had a false-positive rate of 6.3%. Cross-reactivity of the strip was assessed and was negative after tests with clenbuterol analogues such as terbutaline, salbutamol, ractopamine, ritodrine, and fenoterol. These data suggest that a lateral-flow strip assay can be used safely as a screening method as part of a clenbuterol residue surveillance program and should be a valuable tool in the food safety field, especially in developing countries.

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