Simple and rapid detection of Mycobacterium tuberculosis complex organisms in bovine tissue samples by PCR

Liébana, Ernesto; Aranaz, Alicia; Mateos, Ana; Vilafranca, M.; Gómez-Mampaso, E; Tercero, Juan Carlos; Alemany, Jorge; Suárez, G.; Domingo, Mariano Cuesta; Domı́nguez, Lucas

doi:10.1128/jcm.33.1.33-36.1995

Cited by 76 publications

(43 citation statements)

References 16 publications

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“…In conclusion, in a herd infected by the same strain, ante-mortem direct and immune-diagnostic parameters change, suggesting that several tests are needed for a faster control of infection at herd level. detect members of the M. tuberculosis complex and is especially useful for the direct detection of M. bovis in bovine tissue samples (Kolk et al, 1992;Kox et al, 1994;Liebana et al, 1995;Wards et al, 1995;Cornejo et al, 1998;Vitale et al, 1998;Zanini et al, 1998Zanini et al, , 2001Romero et al, 1999;Zumárraga et al, 2001Zumárraga et al, , 2005) Eradication of this disease remains an important goal in several countries. The strategy of test and slaughter has been used widely in an attempt to control dissemination of the disease and is based on the tuberculin skin test as a means for BTB diagnosis.…”

Section: Introductionmentioning

confidence: 99%

Individual Animals of a Cattle Herd Infected with the Same Mycobacterium bovis Genotype Shows Important Variations in Bacteriological, Histopathological and Immune Response Parameters

Meikle¹,

Schneider²,

Azenzo³

et al. 2007

Zoonoses and Public Health

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Cattle are the host and main reservoir of the etiologic agent of bovine tuberculosis, Mycobacterium bovis; although other mammalian species, including humans, are susceptible. The tuberculin test and/or slaughterhouse surveillance is the diagnostic method used by control programs all around the world to control and eradicate the disease. In order to compare different tuberculosis diagnostic tests and to reach disease confirmation, a study was performed in a group of 14 steers of Friesian breed, reacting positively to tuberculin test. Three ante-mortem assays were performed according to the type of sample: the gamma interferon (IFN-gamma) test (which quantifies the release of this cytokine by sensitized lymphocytes in whole blood in response to purified protein derivative (PPD) and recombinant ESAT-6 and CFP10 proteins); PCR and bacteriologic culture from nasal swab and intradermal tuberculin test. These assays were taken at different times to assess the evolution of clinical parameters. Post-mortem examination showed macroscopic and microscopic tuberculosis lesions with acid-fast bacillus and positive cultures. By spoligotyping, we observed that all the isolates showed the same pattern. The positive results based on comparison to lesions observed ranged from 58% to 75% for the IFN-gamma assays, to 72% for cultures, and ranged from 50% to 90% for PCR in nasal swabs. In conclusion, in a herd infected by the same strain, ante-mortem direct and immune-diagnostic parameters change, suggesting that several tests are needed for a faster control of infection at herd level.

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Section: Introductionmentioning

confidence: 99%

Individual Animals of a Cattle Herd Infected with the Same Mycobacterium bovis Genotype Shows Important Variations in Bacteriological, Histopathological and Immune Response Parameters

Meikle¹,

Schneider²,

Azenzo³

et al. 2007

Zoonoses and Public Health

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“…In this study, 27 animals of three species were classified as infected (3 per species) or noninfected (6 per species) on the basis of whether M. bovis could be cultured from tissues. This microbiological procedure is generally considered to be the "gold standard," i.e., the definitive test for the confirmation of M. bovis infection (22,36). The MPB70-based ELISA was performed to select M. bovis-infected animals with anti-MPB70 antibodies and noninfected animals lacking these antibodies ( Table 1).…”

Section: Discussionmentioning

confidence: 99%

Modification of the Mycobacterium bovis extracellular protein MPB70 with fluorescein for rapid detection of specific serum antibodies by fluorescence polarization

Lin¹,

Sugden²,

Jolley³

et al. 1996

Clin Diagn Lab Immunol

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The principle of fluorescence polarization described by Perrin (F. Perrin, J. Phys. Radium 7:390-401, 1926) was applied to the development of a novel assay that used fluorescein-labeled Mycobacterium bovis secretory protein MPB70 for rapid detection of anti-MPB70 antibodies in selected sera from three M. bovis-infected species (elk, llama, and bison). Labeling of purified MPB70 with fluorescein isothiocyanate resulted in the incorporation of 0.96 ؎ 0.08 (mean ؎ standard deviation; n ‫؍‬ 3) fluorescein group per MPB70 molecule. The labeled protein fluoresced strongly with an emission maximum at 518 nm when excited with light of a wavelength near 493 nm, and its immunoreactivity with anti-MPB70 monoclonal antibody 4C3/17 was not altered by modification with fluorescein. The fluorescence polarization assay protocol was optimized for analysis of serum samples by incorporating into the assay buffer 0.05% lithium dodecyl sulfate, which prevents the occurrence of some nonspecific interactions. Sera from M. bovis-infected animals, selected on the basis of exhibiting the presence of anti-MPB70 antibodies, as detected by enzyme-linked immunosorbent assay (ELISA), reacted with fluorescein-labeled MPB70, resulting in an increase in polarization of up to 330 milli-polarization units, in contrast to the values for noninfected sera (167 to 178 mP), which were close to that obtained in the absence of specific antibodies (164.7 ؎ 3.3 mP; n ‫؍‬ 6). These results demonstrated the feasibility of using fluorescein-labeled MPB70 to detect anti-MPB70 antibodies by fluorescence polarization and suggested that the assay described here can be an alternative to ELISA or other antibody assay systems. The advantages of this original methodology and its general applicability to the diagnosis of infectious diseases are discussed.Mycobacterium bovis causes infection (tuberculosis) in a variety of farm and wild animals and also in humans (35). This organism, along with M. tuberculosis, M. africanum, and M. microti, is one of the taxonomically closely related members of the M. tuberculosis complex. Numerous attempts have been made to find a species-specific antigen with high sensitivity for diagnosis of M. bovis infection. The protein antigen MPB70 is secreted from M. bovis cells following cleavage of a 30-aminoacid signal peptide which directs the active transport of the protein across the cytoplasmic membrane (33). The protein forms a major component of M. bovis culture filtrate, accounting for up to 10% of the protein excreted by some M. bovis bacillus Calmette-Guérin (BCG) strains (14,25), and possibly as much as 23% (2). The function of this protein is unknown. The protein was first purified to homogeneity from culture filtrates of M. bovis BCG by Nagai et al. (25) and later was isolated from culture filtrates of M. bovis AN-5 by Fifis et al. (10,11). Also, the gene encoding MPB70 in M. bovis has been cloned, sequenced (28, 33), and expressed in Escherichia coli (18). The molecular mass of MPB70 was estimated to be between 18 and 23 kDa by ...

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“…Culture stands as the 'gold standard' method (M. Miller, 2008) but requires from 2 weeks to 3 months for culture to occur (Liebana et al, 1995), depending on whether the mycobacteria belong to the slow-growing group (e.g. all mycobacteria from MTC) or rapidly growing group (e.g.…”

Section: Limits Of Direct Examinationmentioning

confidence: 99%

Mycobacterial infections in zoo animals: relevance, diagnosis and management*

Lécu¹,

Ball²

2011

International Zoo Yearbook

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While the world prevalence of tuberculosis (TB) is increasing in the human population, TB infection remains a real concern in some animal populations all around the globe. Most mycobacteria of the TB complex are able to infect zoo and wildlife species, in which the pathogenesis, receptivity and immune responses vary widely. The diagnostic tools usually applied in domestic animals show limited performance in zoo species, especially when prevalence is low. Conversely, investigations of cell‐mediated immunity through in vitro assay of γ‐interferon may have numerous advantages, as long as the technical limits are known and can be improved upon. Furthermore, recent tools based on the investigation of humoral immunity seem very promising for the detection of antibodies directed against certain immunogenic mycobacterial antigens in a wide range of species. All these methods are currently evaluated in field studies, despite the difficulties to ensure rigorous validation. The development of these diagnostic tools is also impaired by the prevalence of mycobacteria other than TB also able to infect and create relevant disease in their host. Thus, decisions on positive and suspicious‐animals issues should be taken based on the evaluation of the risk of transmission to the rest of the zoological collection, the possible treatment options, animal welfare, conservation considerations and, of course, the zoonotic potential of this pathogen.

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Simple and rapid detection of Mycobacterium tuberculosis complex organisms in bovine tissue samples by PCR

Cited by 76 publications

References 16 publications

Individual Animals of a Cattle Herd Infected with the Same Mycobacterium bovis Genotype Shows Important Variations in Bacteriological, Histopathological and Immune Response Parameters

Individual Animals of a Cattle Herd Infected with the Same Mycobacterium bovis Genotype Shows Important Variations in Bacteriological, Histopathological and Immune Response Parameters

Modification of the Mycobacterium bovis extracellular protein MPB70 with fluorescein for rapid detection of specific serum antibodies by fluorescence polarization

Mycobacterial infections in zoo animals: relevance, diagnosis and management*

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