scholarly journals Rapid Detection of Serum Antibody by Dual-Path Platform VetTB Assay in White-Tailed Deer Infected with Mycobacterium bovis

2013 ◽  
Vol 20 (6) ◽  
pp. 907-911 ◽  
Author(s):  
Konstantin P. Lyashchenko ◽  
Rena Greenwald ◽  
Javan Esfandiari ◽  
Daniel J. O'Brien ◽  
Stephen M. Schmitt ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Mycobacterium Bovis ◽  
Skin Testing ◽  
Serum Antibody ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Serum Samples ◽  
Content Type ◽  
In The Wild ◽  
Free Ranging

ABSTRACTBovine tuberculosis (TB) in cervids remains a significant problem affecting farmed herds and wild populations. Traditional skin testing has serious limitations in certain species, whereas emerging serological assays showed promising diagnostic performance. The recently developed immunochromatographic dual-path platform (DPP) VetTB assay has two antigen bands, T1 (MPB83 protein) and T2 (CFP10/ESAT-6 fusion protein), for antibody detection. We evaluated the diagnostic accuracy of this test by using serum samples collected from groups of white-tailed deer experimentally inoculated withMycobacterium bovis,M. aviumsubsp.paratuberculosis, orM. bovisBCG Pasteur. In addition, we used serum samples from farmed white-tailed deer in herds with no history of TB, as well as from free-ranging white-tailed deer culled during field surveillance studies performed in Michigan known to have bovine TB in the wild deer population. The DPP VetTB assay detected antibody responses in 58.1% of experimentally infected animals within 8 to 16 weeks postinoculation and in 71.9% of naturally infected deer, resulting in an estimated test sensitivity of 65.1% and a specificity of 97.8%. The higher seroreactivity found in deer with naturally acquiredM. bovisinfection was associated with an increased frequency of antibody responses to the ESAT-6 and CFP10 proteins, resulting in a greater contribution of these antigens, in addition to MPB83, to the detection of seropositive animals, compared with experimentalM. bovisinfection. Deer experimentally inoculated with eitherM. aviumsubsp.paratuberculosisorM. bovisBCG Pasteur did not produce cross-reactive antibodies that could be detected by the DPP VetTB assay. The present findings demonstrate the relatively high diagnostic accuracy of the DPP VetTB test for white-tailed deer, especially in the detection of naturally infected animals.

scholarly journals Effects of Serial Skin Testing with Purified Protein Derivative on the Level and Quality of Antibodies to Complex and Defined Antigens in Mycobacterium bovis-Infected Cattle

2015 ◽  
Vol 22 (6) ◽  
pp. 641-649 ◽  
Author(s):  
W. Ray Waters ◽  
Mitchell V. Palmer ◽  
Molly R. Stafne ◽  
Kristin E. Bass ◽  
Mayara F. Maggioli ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Specific Antibody ◽  
Skin Testing ◽  
Antibody Responses ◽  
Proteinase K ◽  
Skin Tests ◽  
Infected Cattle ◽  
Content Type ◽  
Purified Protein Derivative ◽  
In Series

ABSTRACTSeveral serological tests designed to detect antibodies to immunodominantMycobacterium bovisantigens have recently emerged as ancillary tests for the detection of bovine tuberculosis in cattle, particularly when used after the injection of purified protein derivative (PPD) for skin testing, which significantly boostsM. bovis-specific antibody responses. The present findings demonstrate the onset and duration of boosted antibody responses after the injection ofM. bovisPPD for the caudal fold test (CFT) andMycobacterium aviumandM. bovisPPDs for the comparative cervical test (CCT), administered in series in cattle experimentally infected withM. bovis. While skin tests boosted the responses to certain antigens (i.e., MPB83 and MPB70), they did not affect the responses to other antigens (e.g., ESAT-6, CFP10, MPB59, and MPB64). Administration of the CCT 105 days after the CFT resulted in an even greater secondary boost in antibody responses to MPB83 and MPB70 and to a proteinase K-digested whole-cell sonicate (WCS-PK) ofM. bovis. Both IgM and IgG contributed to the initial boost in the MPB83/MPB70-specific antibody response after the CFT. The secondary boost after the CCT was primarily due to increased IgG levels. Also, the avidity of antibodies to MPB83 and MPB70 increased after the CCT inM. bovis-infected cattle. The avidity of antibodies to the WCS-PK antigens increased in the interval between the CFT and the CCT but did not increase further after the CCT. Together, these findings demonstrate that the administration of PPDs for skin tests results in additive enhancement (i.e., when the CFT and CCT are performed in series), both qualitative and quantitative, of MPB83/MPB70-specific antibody responses.

scholarly journals Detection of Actinobacillus pleuropneumoniae ApxIV toxin antibody in serum and oral fluid specimens from pigs inoculated under experimental conditions

2017 ◽  
Vol 61 (2) ◽  
pp. 163-171 ◽  
Author(s):  
Wendy González ◽  
Luis G. Giménez-Lirola ◽  
Ashley Holmes ◽  
Sergio Lizano ◽  
Christa Goodell ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Serum Antibody ◽  
Population Monitoring ◽  
Actinobacillus Pleuropneumoniae ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Post Inoculation ◽  
Serum Samples ◽  
Experimental Conditions ◽  
And Control

AbstractIntroduction:The prevention and control ofActinobacillus pleuropneumoniaein commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety ofA. pleuropneumoniaeantigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique toA. pleuropneumoniaeand produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.Material and Methods:Four groups of pigs (six pigs per group) were inoculated withA. pleuropneumoniaeserovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.Results:All pigs inoculated withA. pleuropneumoniaeserovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.Conclusion:This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring forA. pleuropneumoniae.

scholarly journals Serum Antibody Responses to 10 Mycobacterium tuberculosis Proteins, Purified Protein Derivative, and Old Tuberculin in Natural and Experimental Tuberculosis in Rhesus Monkeys

2011 ◽  
Vol 18 (12) ◽  
pp. 2154-2160 ◽  
Author(s):  
Fangui Min ◽  
Yu Zhang ◽  
Ren Huang ◽  
Wende Li ◽  
Yu'e Wu ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Rhesus Monkeys ◽  
Skin Testing ◽  
Serum Antibody ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tuberculin Skin Testing ◽  
Purified Protein Derivative ◽  
Diagnostic Potential

ABSTRACTOld tuberculin (OT) and purified protein derivative (PPD) are widely used for tuberculin skin testing (TST) in diagnosis of tuberculosis (TB) but often yield poor specificity and anergy in reaction. Therefore, it is necessary to develop new serological methods as a possible auxiliary diagnostic method for TB. In this study, we characterized the dynamic antibody responses of 10 purified recombinant antigens, PPD, and OT in rhesus monkeys experimentally infected withMycobacterium tuberculosisand analyzed the time to antibody detection, antibody levels, and their association with the infectious doses. The antibodies were detected as early as 4 weeks after infection in response to 5 antigens (CFP10, CFP10-ESAT-6, U1, MPT64, and Ag85b). Antibodies against most of the other antigens were detected between 4 and 12 weeks after infection. The levels of antibodies were dose dependant. We further evaluated the serodiagnostic potential of these antigens by using indirect enzyme-linked immunosorbent assay in 71 TST-positive and 90 TST-negative serum samples from monkeys. For all 12 antigens, the median optical density values of TST-positive monkeys were statistically significantly higher than those of TST-negative monkeys (P< 0.001). Among those antigens, Ag85b and CFP10 showed higher diagnostic potential than others. A combination of results from Ag85b, the 38-kDa antigen (Ag38kDa), and Ag14kDa reaches a sensitivity of 95.77%, indicating that these antigens may be ideal cocktails in TB diagnosis.

scholarly journals Mycobacterium avium subsp. paratuberculosis Antibody Response, Fecal Shedding, and Antibody Cross-Reactivity to Mycobacterium bovis in M. avium subsp. paratuberculosis-Infected Cattle Herds Vaccinated against Johne's Disease

2014 ◽  
Vol 21 (5) ◽  
pp. 698-703 ◽  
Author(s):  
Deepanker Tewari ◽  
Ernest Hovingh ◽  
Rick Linscott ◽  
Edmond Martel ◽  
John Lawrence ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Mycobacterium Avium ◽  
Cross Reactivity ◽  
Johne's Disease ◽  
Antibody Responses ◽  
Johne’S Disease ◽  
Serum Samples ◽  
Content Type ◽  
Fecal Shedding ◽  
Cattle Herds

ABSTRACTVaccination for Johne's disease with killed inactivated vaccine in cattle herds has shown variable success. The vaccine delays the onset of disease but does not afford complete protection. Johne's disease vaccination has also been reported to interfere with measurements of cell-mediated immune responses for the detection of bovine tuberculosis. Temporal antibody responses and fecal shedding ofMycobacterium aviumsubsp.paratuberculosis, the causative agent of Johne's disease, were measured in 2 dairy cattle herds using Johne's disease vaccine (Mycopar) over a period of 7 years. Vaccination against Johne's disease resulted in positive serumM. aviumsubsp.paratuberculosisantibody responses in both herds, and the responses persisted in vaccinated cattle up to 7 years of age. Some vaccinated animals (29.4% in herd A and 36.2% in herd B) showed no serological reactivity toM. aviumsubsp.paratuberculosis.M. aviumsubsp.paratuberculosis-specific antibody responses were also detected in milk from Johne's disease-vaccinated animals, but fewer animals (39.3% in herd A and 49.4% in herd B) had positive results with milk than with serum samples. With vaccination againstM. aviumsubsp.paratuberculosis, fecal shedding in both dairy herds was reduced significantly (P< 0.001). In addition, when selected Johne's disease-vaccinated and -infected animals were investigated for serological cross-reactivity toMycobacterium bovis, no cross-reactivity was observed.

scholarly journals Antibody Responses in Reindeer (Rangifer tarandus) Infected with Mycobacterium bovis

2005 ◽  
Vol 12 (6) ◽  
pp. 727-735 ◽  
Author(s):  
W. R. Waters ◽  
M. V. Palmer ◽  
J. P. Bannantine ◽  
R. Greenwald ◽  
J. Esfandiari ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Specific Antibody ◽  
Rangifer Tarandus ◽  
Skin Testing ◽  
Serum Antibody ◽  
The United States ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cell Mediated Immunity ◽  
Testing Procedures

ABSTRACT Despite having a very low incidence of disease, reindeer (Rangifer tarandus) are subject to tuberculosis (TB) testing requirements for interstate shipment and herd accreditation in the United States. Improved TB tests are desperately needed, as many reindeer are falsely classified as reactors by current testing procedures. Sera collected sequentially from 11 (experimentally) Mycobacterium bovis-infected reindeer and 4 noninfected reindeer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for antibody specific to M. bovis antigens. Specific antibody was detected as early as 4 weeks after challenge with M. bovis. By MAPIA, sera were tested with 12 native and recombinant antigens, which were used to coat nitrocellulose. All M. bovis-infected reindeer developed responses to MPB83 and a fusion protein, Acr1/MPB83, and 9/11 had responses to MPB70. Other antigens less commonly recognized included MPB59, ESAT-6, and CFP10. Administration of purified protein derivatives for skin testing boosted serum antibody responses, as detected by each of the assays. Of the noninfected reindeer, 2/4 had responses that were detectable immediately following skin testing, which correlated with pathological findings (i.e., presence of granulomatous lesions yet the absence of acid-fast bacteria). The levels of specific antibody produced by infected reindeer appeared to be associated with disease progression but not with cell-mediated immunity. These findings indicate that M. bovis infection of reindeer elicits an antibody response to multiple antigens that can be boosted by skin testing. Serological tests using carefully selected specific antigens have potential for early detection of infections in reindeer.

scholarly journals Performance of Commercial Mycoplasma hyopneumoniae Serum Enzyme-Linked Immunosorbent Assays under Experimental and Field Conditions

2020 ◽  
Vol 58 (12) ◽  
Author(s):  
Ana Paula S. Poeta Silva ◽  
Ronaldo L. Magtoto ◽  
Henrique M. Souza Almeida ◽  
Aric McDaniel ◽  
Precy D. Magtoto ◽  
...  
Keyword(s):  
Serum Antibody ◽  
Mycoplasma Hyopneumoniae ◽  
Error Rates ◽  
False Positives ◽  
Field Conditions ◽  
Misclassification Error ◽  
Antibody Detection ◽  
Serum Samples ◽  
Content Type ◽  
Immunosorbent Assays

ABSTRACT Mycoplasma hyopneumoniae is an economically significant pathogen of swine. M. hyopneumoniae serum antibody detection via commercial enzyme-linked immunosorbent assays (ELISAs) is widely used for routine surveillance in commercial swine production systems. Samples from two studies were used to evaluate assay performance. In study 1, 6 commercial M. hyopneumoniae ELISAs were compared using serum samples from 8-week-old cesarean-derived, colostrum-deprived (CDCD) pigs allocated to the following 5 inoculation groups of 10 pigs each: (i) negative control, (ii) Mycoplasma flocculare (strain 27399), (iii) Mycoplasma hyorhinis (strain 38983), (iv) Mycoplasma hyosynoviae (strain 34428), and (v) M. hyopneumoniae (strain 232). Weekly serum and daily oral fluid samples were collected through 56 days postinoculation (dpi). The true status of pigs was established by PCR testing on oral fluids samples over the course of the observation period. Analysis of ELISA performance at various cutoffs found that the manufacturers’ recommended cutoffs were diagnostically specific, i.e., produced no false positives, with the exceptions of 2 ELISAs. An analysis based on overall misclassification error rates found that 4 ELISAs performed similarly, although one assay produced more false positives. In study 2, the 3 best-performing ELISAs from study 1 were compared using serum samples generated under field conditions. Ten 8-week-old pigs were intratracheally inoculated with M. hyopneumoniae. Matched serum and tracheal samples (to establish the true pig M. hyopneumoniae status) were collected at 7- to 14-day intervals through 98 dpi. Analyses of sensitivity and specificity showed similar performance among these 3 ELISAs. Overall, this study provides an assessment of the performance of current M. hyopneumoniae ELISAs and an understanding of their use in surveillance.

scholarly journals Antibody Responses of Cervids (Cervus elaphus) following Experimental Mycobacterium bovis Infection and the Implications for Immunodiagnosis

2008 ◽  
Vol 15 (11) ◽  
pp. 1650-1658 ◽  
Author(s):  
Noel P. Harrington ◽  
Om P. Surujballi ◽  
John F. Prescott ◽  
J. Robert Duncan ◽  
W. Ray Waters ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Cervus Elaphus ◽  
Skin Testing ◽  
Rapid Test ◽  
Antibody Responses ◽  
Tuberculin Skin Testing ◽  
Protein Antigens ◽  
Serum Samples ◽  
Fluorescence Polarization Assay ◽  
Antibody Specificities

ABSTRACT Captive and free-ranging wildlife animals are implicated in the maintenance and transmission of bovine tuberculosis and therefore pose a significant obstacle to eradication of the disease from domestic livestock. The current antemortem diagnostic method, the intradermal tuberculin skin test, is impractical for routine use with many wild animals. Antibody-based assays are particularly attractive because the animals are handled only once and immediate processing of the sample is not required. This report characterizes the antibody responses of red deer-elk hybrids (Cervus elaphus) against Mycobacterium bovis and subsequently evaluates the diagnostic performance of select antigens in a rapid-test format. Sequential serum samples were collected from 10 animals experimentally infected with M. bovis and 5 noninfected animals over a 7-month period postinfection (p.i.). Samples were evaluated by enzyme-linked immunosorbent assays, immunoblot analyses, and multiantigen print immunoassays for seroreactivity to mycobacterial antigens. Although all infected animals produced antibodies to M. bovis protein antigens, there was significant animal-to-animal variation in the kinetics and magnitudes of responses and the antigens recognized. The most frequently recognized antigens included MPB83, ESAT-6, CFP10, and MPB70. Responses to some antigens, such as MPB83, were consistently detected as early as 4 weeks after inoculation, whereas other antigens were detected only much later (>140 days p.i.). Antibody responses were boosted by injection of tuberculin for intradermal tuberculin skin testing. Comparison of single-antigen (fluorescence polarization assay) with multiantigen (CervidTB STAT-PAK) rapid tests demonstrated that a highly sensitive and specific serodiagnostic test for tuberculosis in cervids will require multiple and carefully selected seroreactive antigens covering a broad spectrum of antibody specificities.

scholarly journals Identification of Novel Antigens Recognized by Serum Antibodies in Bovine Tuberculosis

2017 ◽  
Vol 24 (12) ◽  
Author(s):  
Konstantin P. Lyashchenko ◽  
Adrian Grandison ◽  
Karen Keskinen ◽  
Alina Sikar-Gang ◽  
Paul Lambotte ◽  
...  
Keyword(s):  
Immune Responses ◽  
Diagnostic Accuracy ◽  
Mycobacterium Bovis ◽  
Recombinant Proteins ◽  
Bovine Tuberculosis ◽  
Serum Antibodies ◽  
Serum Samples ◽  
Content Type ◽  
Humoral Immune ◽  
Humoral Immune Responses

ABSTRACT Bovine tuberculosis (TB), caused by Mycobacterium bovis, remains an important zoonotic disease posing a serious threat to livestock and wildlife. The current TB tests relying on cell-mediated and humoral immune responses in cattle have performance limitations. To identify new serodiagnostic markers of bovine TB, we screened a panel of 101 recombinant proteins, including 10 polyepitope fusions, by a multiantigen print immunoassay (MAPIA) with well-characterized serum samples serially collected from cattle with experimental or naturally acquired M. bovis infection. A novel set of 12 seroreactive antigens was established. Evaluation of selected proteins in the dual-path platform (DPP) assay showed that the highest diagnostic accuracy (∼95%) was achieved with a cocktail of five best-performing antigens, thus demonstrating the potential for development of an improved and more practical serodiagnostic test for bovine TB.

scholarly journals Early Antibody Responses to Experimental Mycobacterium bovis Infection of Cattle

2006 ◽  
Vol 13 (6) ◽  
pp. 648-654 ◽  
Author(s):  
W. R. Waters ◽  
M. V. Palmer ◽  
T. C. Thacker ◽  
J. P. Bannantine ◽  
H. M. Vordermeier ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Skin Testing ◽  
Rapid Test ◽  
Kinetic Studies ◽  
Immunoblot Analysis ◽  
Immunoglobulin M ◽  
Serum Samples ◽  
Mycobacterial Antigens ◽  
New Generation ◽  
And Control

ABSTRACT Bovine tuberculosis persists as a costly zoonotic disease in numerous countries despite extensive eradication and control efforts. Sequential serum samples obtained from Mycobacterium bovis-infected cattle were evaluated for seroreactivity to mycobacterial antigens. Animals received M. bovis by aerosol, intratonsil, intranasal, or intratracheal inoculation. Assays included the multiantigen print immunoassay for determination of antigen recognition patterns, immunoblot analysis for sensitive kinetic studies, and the VetTB STAT-PAK test, a novel, rapid test based on lateral-flow technology. Responses to MPB83 were detected for all M. bovis-infected animals regardless of the route or strain of M. bovis used for inoculation. Other less commonly recognized antigens included ESAT-6, CFP-10, and MPB70. Responses to MPB83 were detectable as early as 4 weeks after inoculation, were boosted upon injection of purified protein derivatives for skin testing, and persisted throughout the course of each of the four challenge studies. MPB83-specific immunoglobulin M (IgM) was detected prior to MPB83-specific IgG detection; however, early IgM responses rapidly waned, suggesting a benefit of tests that detect both IgM- and IgG-specific antibodies. The VetTB STAT-PAK test detected responses in sera from 60% (15/25) of the animals by 7 weeks after challenge and detected responses in 96% (24/25) of the animals by 18 weeks. These findings demonstrate the potential for new-generation antibody-based tests for the early detection of M. bovis infection in cattle.

scholarly journals Help in the Choice of Automated or Semiautomated Immunoassays for Serological Diagnosis of Toxoplasmosis: Evaluation of Nine Immunoassays by the French National Reference Center for Toxoplasmosis

2016 ◽  
Vol 54 (12) ◽  
pp. 3034-3042 ◽  
Author(s):  
O. Villard ◽  
B. Cimon ◽  
C. L'Ollivier ◽  
H. Fricker-Hidalgo ◽  
N. Godineau ◽  
...  
Keyword(s):  
Congenital Toxoplasmosis ◽  
Antibody Detection ◽  
Clinical Settings ◽  
Serum Samples ◽  
Igg Antibody ◽  
Routine Testing ◽  
Content Type ◽  
Specificity And Sensitivity ◽  
National Reference Center ◽  
Serology Testing

Toxoplasmosis, a benign infection, is asymptomatic or paucisymptomatic in over 80% of cases, except in immunocompetent patients suffering from ocular toxoplasmosis or in immunocompromised patients with opportunistic or congenital toxoplasmosis. Diagnosis is based mainly on serology testing. Thus, we compared the performance of the nine most commonly used commercial automated or semiautomated immunoassays for IgG and IgMToxoplasma gondiiantibody detection, that is, the Advia Centaur, Architect, AxSYM, Elecsys, Enzygnost, Liaison, Platelia, VIDAS, and VIDIA assays. The assays were conducted on four panels of serum samples derived during routine testing from patients with an interfering disease and who exhibited a low IgG antibody level in one of two clinical settings, namely, acute or chronic toxoplasmosis. As a result, IgG sensitivities ranged from 97.1% to 100%, and IgG specificities ranged from 99.5% to 100%. For IgG quantification, strong differences in IgG titers (expressed in IU/ml) were noted depending on the assay used. IgM sensitivities ranged from 65% to 97.9%, and IgM specificities ranged from 92.6% to 100%. For defining the best serological strategies to be implemented, it appears crucial to compare the diagnostic performance of the different tests with respect to their specificity and sensitivity in detecting the presence of IgG and IgM antibodies.

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