Identification of Mycobacterium bovis Isolates by a multiplex PCR

Abstract

Isolates from suggestive bovine tuberculosis lesions were tested by a multiplex polymerase chain reaction (m-PCR) targeting for RvD1Rv2031c and IS6110 sequences, specific for M. bovis and Mycobacterium tuberculosis complex respectively. The m-PCR successfully identified as M. bovis 88.24% of the isolates.

A multidisciplinary approach to diagnose naturally occurring bovine tuberculosis in Brazil

“…It has also been reported that different strains of M.bovis could enhance different subclasses of IgG (Jolley et al 2007), a factor that could contribute to the variation seen in ELISA results. Since the selection of an antigen for any assay is one of the most important steps in the development of that assay, the techniques by which that test is used can also be a determinant toward achieving higher levels of sensitivity and specificity (Jolley et al 2007, Figueiredo et al 2008. Nevertheless, the unexpected lack of a booster effect in our study remains unexplained.…”

“…To culture M. bovis, samples of tissues, milk and swabs were decontaminated according to standard methods (Lilenbaum 1999) and inoculated onto two slopes of solid, egg-based media Lowenstein-Jensen with 0.5% pyruvate, and two slopes of Stonebrink media, which were incubated at 37 o C and observed once weekly for 12 weeks. For evidence of M. bovis-specific DNA, a multiplex PCR targeting the RvD1Rv2031c genomic sequence was employed (Figueiredo et al 2008).…”

“…Therefore, it was published as short communications about multiplex PCR (Figueiredo et al 2008), IFN (Marassi et al 2009b) and ELISA (Lilenbaum et al 2011). In this article, we propose a comprehensive discussion of the information obtained during the project, analysing the various diagnostic tools as a conjunct.…”

Detection of specific antibodies in cows after injection of PPD

“…At necropsy, although only two of them presented characteristic gross lesions at necropsy, such as tubercles with central caseation and calcification, or circumscribed yellowish white lesions in lungs or lymph nodes. From the nine cows mediastinal, bronchial lymph nodes, and lung samples were collected and processed for culture and PCR (Figueiredo et al 2009). …”

Diagnóstico histopatológico e molecular de lesões sugestivas de tuberculose em búfalos abatidos nos municípios de Macapá e Santana, estado do Amapá

“…Em bovinos reagentes à tuberculinização, Figueiredo et al (2009) identificaram M. bovis em 88,24% (15/17) das amostras, utilizando dois pares de primers específicos para M. bovis e M. tuberculosis, por meio de PCR múltipla. Parra et al (2008), desenvolvendo sistema de PCR múltipla em tempo real, diretamente de amostras biológicas, obtiveram taxas de sensibilidade variando de 61,11% (lesões não visíveis) até 80,64% para lesões tuberculosas crônicas, permitindo a diferenciação entre membros do CMA e outras micobactérias não-tuberculosas.…”