Evaluation of two cocktails containing ESAT-6, CFP-10 and Rv-3615c in the intradermal test and the interferon-γ assay for diagnosis of bovine tuberculosis

Casal, Carmen; Bezos, Javier; Díez-Guerrier, Alberto; Álvarez, Julio; Romero, Beatriz; Juan, Lucía de; Rodriguez‐Campos, Sabrina; Vordermeier, Martin; Whelan, Adam O.; Hewinson, R. Glyn; Mateos, Ana; Domı́nguez, Lucas; Aranaz, Alicia

doi:10.1016/j.prevetmed.2012.02.007

Cited by 43 publications

(59 citation statements)

References 17 publications

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“…Although the gamma interferon (IFN-␥) release assay (IGRA) showed higher specificity than the single intradermal skin test, the high price of this kit limits its use in developing countries such as China (9, 10). To overcome the drawbacks of the traditional diagnostic methods, scientists have focused on screening new M. bovis-specific antigens to substitute for PPD-B as stimuli in the skin test (11)(12)(13).Recent studies focused mainly on the development of skin test reagents to discriminate infected from vaccinated cattle (DIVA test), in which M. bovis-specific antigens, such as CFP10 and ESAT-6, were often selected as stimuli for the skin test (6,11,(14)(15)(16). Those antigens were located in RD1, deleted in M. bovis BCG strains, and could elicit strong DTH in TB-infected cattle.…”

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confidence: 99%

Assessment of a Protein Cocktail-Based Skin Test for Bovine Tuberculosis in a Double-Blind Field Test in Cattle

Xin

Ding

et al. 2013

Clin Vaccine Immunol

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Bovine tuberculosis (bTB) is a worldwide zoonosis caused mainly by Mycobacterium bovis. The traditional diagnostic method used often is the tuberculin skin test, which uses bovine purified protein derivatives (PPD-B). However, it is difficult to maintain uniformity of PPD-B from batch to batch, and it shares common antigens with nonpathogenic environmental mycobacteria. To overcome these problems, M. bovis-specific antigens that showed good T cell stimulation, such as CFP-10, ESAT-6, Rv3615c, etc., have been used in the skin test, but there have been no large-scale clinical studies on these antigens. In this study, two combinations (CFP-10/ESAT-6/TB10.4 protein cocktail and CFP-10/ESAT-6/Rv3872/MPT63 protein cocktail) were developed and used as stimuli in the skin test. Cattle were double-blind tested to assess the efficiency of the protein cocktail-based skin tests. The results showed that the CFP-10/ESAT-6/TB10.4 protein cocktail-based skin test can differentiate TB-infected cattle from Mycobacterium avium-infected ones and that it shows a high degree of agreement with the traditional tuberculin skin test ( ‫؍‬ 0.8536) and gamma interferon (IFN-␥) release assay ( ‫؍‬ 0.8154). Compared to the tuberculin skin test, the relative sensitivity and relative specificity of the CFP-10/ESAT-6/TB10.4-based skin test were 87% and 97%, respectively., The relative sensitivity and relative specificity of the CFP-10/ESAT-6/TB10.4-based skin test were 93% and 92%, respectively, on comparison with the IFN-␥ release assay. The correlation between the increases in skin thickness observed after the inoculation of stimuli was high ( (1, 2). M. bovis mainly infects cattle, causing bovine tuberculosis (bTB), and occasionally affects other species of mammals (3). M. bovis can also infect humans through the inhalation of aerosols or the ingestion of unpasteurized dairy products (4, 5). The eradication of bovine tuberculosis is based on reliable diagnostic methods.The traditional diagnostic method of identifying bTB is the tuberculin skin test, which is based on the detection of delayedtype hypersensitivity (DTH) induced by bovine purified protein derivative (PPD-B). The tuberculin skin test has been used worldwide and is economical and easily applied. However, PPD-B is a poorly defined mixture of proteins, lipids, and carbohydrates obtained from a virulent M. bovis culture, the uniformity of which is difficult to maintain from batch to batch (6). Moreover, the use of virulent M. bovis during the production of PPD-B may potentially harm workers. Importantly, shared antigenic components in PPD-B and nonpathogenic environmental mycobacteria can reduce the specificity of the tuberculin skin test (7,8). Although the gamma interferon (IFN-␥) release assay (IGRA) showed higher specificity than the single intradermal skin test, the high price of this kit limits its use in developing countries such as China (9, 10). To overcome the drawbacks of the traditional diagnostic methods, scientists have focused on screening new M. bovis-specific antige...

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confidence: 99%

Assessment of a Protein Cocktail-Based Skin Test for Bovine Tuberculosis in a Double-Blind Field Test in Cattle

Xin

Ding

et al. 2013

Clin Vaccine Immunol

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“…obtained by the use of ESAT-6 and cocktails combinations of ESAT-6 and CFP-10 in previous studies , Whelan et al 2003, Casal et al 2012, there is no sufficient evidence to date to support the replacement of traditional PPD by these recombinant proteins, showing that it is necessary to evaluate other proteins as antigens.…”

Section: Resultsmentioning

confidence: 97%

“…In previous assessments of recombinant proteins as antigens in skin test in cattle, only experimentally infected , Xin et al 2013 or naturally infected , Casal et al 2012 animals had been used. In the present study, for the first time, sensitised animals were used.…”

Section: Resultsmentioning

confidence: 99%

“…Significant differences in the magnitude of the reactions of M. bovis PPD and recombinant proteins have been reported in previous studies , and consequently cut-off points of few millimeters have been utilized to interpret recombinant proteins or synthetic peptide skin tests , Jones et al 2010, Casal et al 2012. In the present study, the median of increase in skin fold thickness in the reactions to the Mb0143 antigen was only 0.5 mm (interquartile range = 0.25 to 1), whereas median values were 1.2 mm (interquartile range = 0.87 and 1.82) for EsxI and 1.95 mm (interquartile range = 1.15 to 2.52) for PE5.…”

Section: Resultsmentioning

confidence: 99%

“…However, only some of them have been evaluated as antigens in the skin test on cattle as an alternative to traditional PPD, such as ESAT-6 , Flores-Villalva et al 2012, CFP-10, MPB70 and MPB83 . Although promising results have been observed with the use of ESAT-6 and CFP-10 either individually or in cocktail combinations , Casal et al 2012, there is no sufficient evidence to date to support the replacement of traditional PPD with these recombinant proteins, mainly due to problems regarding sensitivity . Jones et al (2010) reported a sensitivity level of 87.5% and specificity of 97.6% using a protein cocktail of ESAT-6/ CFP-10/Rv-3615c with the addition of Rv3020c.…”

Section: Introductionmentioning

confidence: 99%

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High-yield production of major T-cell ESAT6-CFP10 fusion antigen of M. tuberculosis complex employing codon-optimized synthetic gene

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Evaluation of two cocktails containing ESAT-6, CFP-10 and Rv-3615c in the intradermal test and the interferon-γ assay for diagnosis of bovine tuberculosis

Cited by 43 publications

References 17 publications

Assessment of a Protein Cocktail-Based Skin Test for Bovine Tuberculosis in a Double-Blind Field Test in Cattle

Assessment of a Protein Cocktail-Based Skin Test for Bovine Tuberculosis in a Double-Blind Field Test in Cattle

Evaluation of the use of recombinant proteins of Mycobacterium bovis as antigens in intradermal tests for diagnosis of bovine tuberculosis

High-yield production of major T-cell ESAT6-CFP10 fusion antigen of M. tuberculosis complex employing codon-optimized synthetic gene

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