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 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 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.

Blood-based assays to detect Mycobacterium bovis-infected cattle missed by tuberculin skin testing

2008 ◽  
Vol 162 (12) ◽  
pp. 382-383 ◽  
Author(s):  
M. Coad ◽  
S. H. Downs ◽  
P. A. Durr ◽  
R. S. Clifton-Hadley ◽  
R. G. Hewinson ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Skin Testing ◽  
Tuberculin Skin Testing ◽  
Infected Cattle

scholarly journals Tuberculin Skin Testing Boosts Interferon Gamma Responses to DIVA Reagents in Mycobacterium bovis-Infected Cattle

2017 ◽  
Vol 24 (5) ◽  
Author(s):  
Gareth J. Jones ◽  
Mick Coad ◽  
Bhagwati Khatri ◽  
Javier Bezos ◽  
Natalie A. Parlane ◽  
...  
Keyword(s):  
Skin Test ◽  
Mycobacterium Bovis ◽  
Interferon Gamma ◽  
Skin Testing ◽  
Interferon Response ◽  
Tuberculin Skin Testing ◽  
Test Scenario ◽  
Positive Skin ◽  
Content Type ◽  
Bcg Vaccination

ABSTRACT Mycobacterium bovis BCG vaccination sensitizes cattle to bovine tuberculin, which compromises the use of the current bovine tuberculosis (TB) surveillance tests. Although the performance of a blood test (that utilizes antigens expressed by Mycobacterium bovis but not by BCG) capable of discriminating infected from vaccinated animals (DIVA interferon gamma test [DIT]) has been evaluated in naturally infected TB field reactors, there is a need to perform similar analysis in a BCG-vaccinated M. bovis-infected population. Furthermore, we explored different scenarios under which a DIT may be implemented alongside BCG vaccination: (i) serial testing to resolve potential false-positive skin test results or (ii) a standalone test to replace the single intradermal comparative cervical tuberculin (SICCT) skin test. Our results demonstrated significantly better relative test sensitivity when the DIT was evaluated in a serial test scenario. Direct comparison of pre- and post-skin test blood samples revealed that the SICCT test induced significant boosting of the gamma interferon response in M. bovis-infected animals to both the ESAT-6–CFP-10 and Rv3615c peptide cocktails that comprise the DIT, which persisted for the ESAT-6–CFP-10 reagent for at least 14 days. Importantly, no similar boosting effects were observed in noninfected BCG vaccinates, suggesting that DIVA blood testing after a recent skin test would have minimal impact on test specificity.

scholarly journals Added Value of Use of a Purified Protein Derivative-Based Enzyme-Linked Immunosorbent Spot Assay for Patients with Mycobacterium bovis BCG Infection after Intravesical BCG Instillations

2012 ◽  
Vol 19 (6) ◽  
pp. 974-977 ◽  
Author(s):  
Karen A. Heemstra ◽  
Ailko W. J. Bossink ◽  
Roan Spermon ◽  
John J. M. Bouwman ◽  
Robert van der Kieft ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Immune Activation ◽  
Bladder Carcinoma ◽  
Case Series ◽  
Gamma Interferon ◽  
Added Value ◽  
Content Type ◽  
Mycobacterium Bovis Bcg ◽  
Purified Protein Derivative ◽  
Intravesical Bcg

ABSTRACTIn this case series, we describe four cases in which the use of gamma interferon release assays with purified protein derivative (PPD) as a stimulating antigen was able to demonstrate PPD-specific immune activation. This may help to improve the adequate diagnosis of (systemic)Mycobacterium bovisBCG infections after intravesical BCG instillations for bladder carcinoma.

scholarly journals Risk Factors for Tuberculosis Conversion in a State Prison

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Robert Hung ◽  
Steven Shelton ◽  
Gary Rischitelli
Keyword(s):  
Risk Factors ◽  
Conversion Rate ◽  
Skin Testing ◽  
Latent Tuberculosis ◽  
Skin Tests ◽  
Purified Protein Derivative ◽  
Medical Chart Review ◽  
Prison Health ◽  
Male Inmates ◽  
Control Study

A case-control study determined the risk factors for latent tuberculosis (TB) conversion among Oregon Department of Correction (ODOC) inmates from July 2000 - July 2001. The first objective was to identity the converters. These were inmates who tested negative for the Purified Protein Derivative (PPD) skin test on entry and subsequently tested positive on annual testing. The second objective was determining the risk factors for conversion by comparing the converters with randomly selected controls. The Correctional Information System (CIS) and Mental Health databases were accessed to obtain health and demographic information. With ninety-nine percent of PPD positive inmates on anti-tuberculosis medications, nearly all male inmates who tested positive from July 00-01 (n = 307) were identified through the ODOC pharmacy records. A medical chart review (276 of 307 or 90%) separated the converters (n = 72) from the reactors who tested positive on entry (n = 123) and the prior positives on medications (n = 81). The conversion rate was 5.0 per 1,000 person-years. Differences between the cases (converters) and controls were analyzed using multivariate logistic regression. The converters were 6 times more likely to be Latino (p < .005) vs. Caucasian, over 19 times less likely to live in medium vs. minimum (p < .001) or maximum vs. minimum (p < .001) security prisons, and over 5 times less likely to live in a medium vs. low (.012 < p < .031) or high vs. low (.002 < p < .007) density prison. They had 1.4-1.5 times fewer PPD skin tests (.002 < p < .009) and lived in 1.5-1.7 times fewer prisons (.005 < p < .017). Age, education, county of incarceration, number of incarcerations, and number of visitors were not found to be significant variables. The results revealed a low conversion rate compared to other U.S. prisons. Prison health officials should consider performing two-step skin testing in order to distinguish the booster phenomenon from intramural conversion.

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 Evaluation of Gamma Interferon (IFN-γ)-Induced Protein 10 Responses for Detection of Cattle Infected with Mycobacterium bovis: Comparisons to IFN-γ Responses

2012 ◽  
Vol 19 (3) ◽  
pp. 346-351 ◽  
Author(s):  
W. R. Waters ◽  
T. C. Thacker ◽  
B. J. Nonnecke ◽  
M. V. Palmer ◽  
I. Schiller ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Similar Pattern ◽  
Gamma Interferon ◽  
Content Type ◽  
Purified Protein Derivative ◽  
Testing Protocol ◽  
Archived Samples ◽  
Ifn Γ ◽  
Highly Correlated ◽  
Mycobacterial Antigens

ABSTRACTGamma interferon (IFN-γ)-induced protein 10 (IP-10) has recently shown promise as a diagnostic biomarker ofMycobacterium tuberculosisinfection of humans. The aim of the current study was to compare IP-10 and IFN-γ responses uponMycobacterium bovisinfection in cattle by using archived samples from two aerosol inoculation studies. In the first study (104CFUM. bovisby aerosol,n= 7),M. bovispurified protein derivative (PPDb)-specific IP-10 and IFN-γ gene expression was detected as early as 29 days after challenge. PPDb-specific IP-10 and IFN-γ mRNA responses followed a similar pattern of expression over the course of this study and were highly correlated (r= 0.87). In the second study (105CFUM. bovisby aerosol,n= 5), IP-10 and IFN-γ (protein) responses to mycobacterial antigens were compared following challenge. IFN-γ responses to mycobacterial antigens were detected at 29 days after challenge and were sustained during the remainder of the study. IFN-γ responses to mycobacterial antigens exceeded corresponding responses in nonstimulated cultures. IP-10 responses to mycobacterial antigens exceeded preinfection responses at 7, 29, and 63 days after challenge. In contrast to IFN-γ responses, IP-10 responses to mycobacterial antigens generally did not exceed the respective responses in nonstimulated cultures. IP-10 responses to medium alone and to mycobacterial antigens followed a similar pattern of response. Correlations between IP-10 and IFN-γ (protein) responses were modest (r≈ 0.50 to 0.65). Taken together, these findings do not support the use of IP-10 protein as a biomarker for bovine tuberculosis using the current testing protocol and reagents; however, mRNA-based assays may be considered for further analysis.

scholarly journals Use of Antigen-Specific Interleukin-2 To Differentiate between Cattle Vaccinated with Mycobacterium bovis BCG and Cattle Infected with M. bovis

2013 ◽  
Vol 21 (1) ◽  
pp. 39-45 ◽  
Author(s):  
Shelley G. Rhodes ◽  
Lucy C. McKinna ◽  
Sabine Steinbach ◽  
Gilly S. Dean ◽  
Bernardo Villarreal-Ramos ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Whole Blood ◽  
Sandwich Elisa ◽  
Interleukin 2 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mycobacterial Antigen ◽  
Content Type ◽  
Purified Protein Derivative ◽  
Rapid Induction ◽  
Challenge Control

ABSTRACTWe describe here the application of a novel bovine interleukin-2 (IL-2) enzyme-linked immunosorbent assay (ELISA) for the measurement of antigen-specific IL-2 in cattle naturally infected withMycobacterium bovisand in cattle vaccinated withMycobacterium bovisBCG and then experimentally challenged with pathogenicM. bovis. Supernatants from whole-blood cultures stimulated with mycobacterial antigen (bovine purified protein derivative [PPDB] or the peptide cocktail ESAT6-CFP10) were assessed using a sandwich ELISA consisting of a new recombinant monoclonal fragment capture antibody and a commercially available polyclonal anti-bovine-IL-2. The production of IL-2 was compared to the production of gamma interferon (IFN-γ) in the same antigen-stimulated whole-blood supernatants. The data show that cattle infected withM. bovisproduced quantifiable levels of antigen-specific IL-2, while IL-2 levels in cattle vaccinated withM. bovisBCG did not. Furthermore, cattle vaccinated withM. bovisBCG and then challenged with pathogenicM. bovisdisplayed a more rapid induction of IL-2 but ultimately had lower levels of infection-induced IL-2 than did unvaccinated challenge control cattle. These data suggest that IL-2 responses are not detectable post-BCG vaccination and that these responses may require infection with virulentM. bovisto develop. This may be useful to differentiate infected cattle from uninfected or BCG-vaccinated cattle, although the overall sensitivity is relatively low, particularly in single intradermal comparative cervical tuberculin (SICCT)-negative infected animals. Furthermore, the strength of the IL-2 response may correlate with pathology, which poses interesting questions on the immunobiology of bovine tuberculosis in contrast to human tuberculosis, which is discussed.

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