Optimizing Antigen Cocktails for Detection of<i>Mycobacterium bovis</i>in Herds with Different Prevalences of Bovine Tuberculosis: ESAT6-CFP10 Mixture Shows Optimal Sensitivity and Specificity

Aagaard, Claus; Govaerts, Marc; Meikle, Virginia; Vallecillo, Antonio J.; Gutiérrez‐Pabello, José A.; Suárez-Güemes, Francisco; McNair, J.; Cataldi, Angel; Espitia, Clara; Andersen, Peter; Pollock, J.M.

doi:10.1128/jcm.01184-06

Cited by 76 publications

(68 citation statements)

References 39 publications

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“…Previous studies have reported that Ag85A, Rv3881c, LAM, Ara 6-BSA and ESAT-6 may be useful for the serodiagnosis of pulmonary and bovine TB respectively [7][8][9][10][11]. Ag85A, ESAT-6 and TB10.4 are strongly recognized by T-cells isolated from TB patients and have been seen to induce a robust cell mediated immune response [7,12,13]. However, little is known about the diagnostic potential of these antigens in EPTB.…”

Section: Introductionmentioning

confidence: 99%

Serodiagnosis of tuberculous lymphadenitis using a combination of antigens

Beyene

Franken

Yamuah

et al. 2010

J Infect Dev Ctries

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Background: The diagnosis of extra-pulmonary tuberculosis (EPTB) by conventional methods such as culture and microscopy has low sensitivity and requires an invasive procedure. A simple rapid serological test would be of great value. Methodology: Six antigens (ESAT-6, Ag85A, TB10.4, Rv3881c, lipoarabinomannan (LAM) and Ara 6 -BSA) were tested in an ELISA to detect antigen-specific IgG and IgM antibodies in sera from 54 culture-and histology-confirmed tuberculous lymphadenitis (TBLN) patients as follows: four were HIV seropositive; sera from 25 was smear positive for pulmonary tuberculosis (PTB); 15 were culture-and histologynegative lymphadenitis (non-TBLN) patients; and 22 werehealthy controls (HCs). Results: The sensitivities of the antigens for the detection of IgG in sera of TBLN patients ranged from 4% to 30%. Specificities ranged from 73% to 100% with sera from non-TBLN patients and 91% to 100% with sera from HCs. Sensitivities of the antigens for detection of IgM ranged from 0% to 15% and specificities ranged from 80% to 100% with sera from non-TBLN patients and 91% to 100% with sera from HCs. LAM was the most potent antigen for detection of IgG. When LAM and ESAT-6 were combined, sensitivity increased up to 43% and specificity with non-TBLN was 80% with HC 96%. Conclusions: The study suggests that the combined use of LAM and ESAT-6 for IgG antibody detection in sera from TBLN patients could be a supplement to microscopy of fine-needle aspirate (FNA) to diagnose TBLN among patients suspected of TBLN.

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Section: Introductionmentioning

confidence: 99%

Serodiagnosis of tuberculous lymphadenitis using a combination of antigens

Beyene

Franken

Yamuah

et al. 2010

J Infect Dev Ctries

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“…Sin embargo, la exposición a otras micobacterias o la co-infección con M. avium subsp paratuberculosis, afecta la especificidad de las pruebas basadas en el uso del PPD, incrementando el número de reacciones falsas positivas, fenómeno que puede ser evitado usando antígenos altamente inmunogénicos como ESAT-6 y CFP-10 (23) . En el presente estudio se observó que el cóctel proteico E6-C10 identificó más animales infectados que el PPDB en la prueba de IFN-γ, el 72.7 % (n= 8) de los animales se identificaron positivamente por el cóctel proteico E6-10, mientras que solo el 36.3 % (n= 4) por el PPDB (15,24) . Estas diferencias se atribuyen principalmente a la pobre definición de antígenos presentes en el PPDB, pues aunque éste proporciona una excelente sensibilidad, su especificidad es limitada, debido a que muchos antígenos presentes en el PPDB también se encuentran presentes en micobacterias ambientales.…”

Section: Cuadro 1 Bovinos Identificados En La Prueba De Ifn-γ Usandounclassified

“…Estas diferencias se atribuyen principalmente a la pobre definición de antígenos presentes en el PPDB, pues aunque éste proporciona una excelente sensibilidad, su especificidad es limitada, debido a que muchos antígenos presentes en el PPDB también se encuentran presentes en micobacterias ambientales. Por otro lado, el coctel proteico E6-C10 está formado por dos antígenos que solo están presentes en bacterias del complejo M. tuberculosis, pero están ausentes en todas las cepas de M. bovis BCG y micobacterias ambientales, por lo que la especificidad de este cóctel es mucho mayor (15,17,24) .…”

Section: Cuadro 1 Bovinos Identificados En La Prueba De Ifn-γ Usandounclassified

“…Los resultados se expresaron usando un índice de densidad óptica (IDO) que se define como el radio de la DO de los cultivos estimulados con un antígeno entre la DO de cultivos estimulados sin antígeno. Para todos los antígenos probados se seleccionó un valor IDO ≥2 como punto de corte para indicar un resultado positivo (15) . Para confirmar la infección por M. bovis en los bovinos, se identificó el material genómico de la micobacteria en las muestras de exudado nasal, las cuales se colectaron utilizando hisopos estériles sumergidos en 2 ml de PBS 1x estéril.…”

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Mycobacterium bovis induce una respuesta inmune celular heterogénea en bovinos infectados naturalmente

López¹,

Flores-Villalva

Pabello³

2017

Rev. Mex. Cienc. Pecu.

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la hipótesis de que las limitaciones en el funcionamiento de los macrófagos y los linfocitos T desempeñan un papel muy importante en la diseminación de la enfermedad.Palabras clave: IFN-γ, Macrófagos, Mycobacterium bovis, PPDB, Coctel proteico. Abstract:Macrophages are considered one of the main immune mechanisms of resistance to mycobacterial infections. In addition, experimental and clinical data indicate that IFN-γ is essential for host defence against M. tuberculosis. The aim of this study was to identify macrophage capacity on controlling Mycobacterium bovis infection and lymphocyte IFN-γ production from cattle naturally exposed to M. bovis. Peripheral blood monocyte-derive macrophages from naturally infected cattle were evaluated for their ability to control the intracellular growth of M. bovis BCG. Besides, whole blood cells were stimulated with PPDB and ESAT-6+CFP-10 protein cocktail. Results show a disparity in host susceptibility to Mycobacterium bovis. A M. bovis BCG survival rate of 80 to 150 % in macrophages from naturally infected cattle was observed. While the IFN-γ production towards Purified protein derivative bovine (PPDB) and the protein cocktail was 2.5 and 2.0 optic density index, respectively. Though our data do not support a correlation between the host susceptibility to M. bovis and the production of IFN-γ in naturally infected cattle, the hypothesis that the limitations in the functioning of macrophages and T lymphocytes play a very important role in the spread of the disease cannot be discarded.

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“…Based on previous reports of diagnosis of tuberculosis in cattle using IFN-γ assays (EsxI, Mb0143, PE5, PE13 and TB10.4) (Aagaard et al 2006, Jones et al 2010, Meikle et al 2009, the aim of the present study was to assess these recombinant proteins with respect to the ability to differentiate M. bovis sensitised and non-sensitised cattle by skin test, for further evaluation of the best antigens in infected animals.…”

Section: Introductionmentioning

confidence: 99%