Otimização da técnica da PCR para a detecção de Actinobacillus pleuropneumoniae

Souza, Karina Koerich de; Klein, Cátia Silene; Kich, Jalusa Deon; Coldebella, Arlei; Alberton, Geraldo Camilo

doi:10.1590/s0103-84782008000800023

Cited by 1 publication

(1 citation statement)

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“…The apxIVA gene and its product have been frequently used as one of the most important molecular markers in the detection and serodiagnosis of A. pleuropneumoniae (Dreyfus et al, 2004;Yang et al, 2009;Urbaniak & Markowska-Daniel, 2011). The omlA and cpx genes have also been used as markers for A. pleuropneumoniae, but we opted not to use these genes in the present work due to serotype variability and previous problems with nonspecific amplifications, respectively (Gram et al, 2000;Souza et al, 2008). When using the primer pair APXIV-Up and APXIV-Do and the amplification conditions suggested by Costa et al (2004), only 84% of our clinical isolates, which had been previously identified by biochemical tests and characterised by multiplex PCR (Rossi et al, 2013), were positive for the apxIVA DNA fragment.…”

Section: Resultsmentioning

confidence: 99%

A BOX-SCAR fragment for the identification ofActinobacillus pleuropneumoniae

Rossi

Pereira

Langford

et al. 2014

FEMS Microbiol Lett

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Bacterial respiratory diseases are responsible for considerable mortality, morbidity and economic losses in the swine industry. Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is one of the most important disease agents, but its identification and surveillance can be impaired by the existence of many other related bacteria in normal swine microbiota. In this work, we have evaluated a BOX-A1R-based repetitive extragenic palindromic-PCR (BOX-PCR) sequence characterised amplified region (SCAR) marker for the specific identification of A. pleuropneumoniae and its use in a multiplex PCR to detect additionally Haemophilus parasuis and Pasteurella multocida, two other major respiratory pathogens of pigs that are members of the family Pasteurellaceae. PCRs based on the BOX-SCAR fragment developed were rapid, sensitive and differentiated A. pleuropneumoniae from all swine-related members of the Pasteurellaceae family tested. Single and multiplex BOX-SCAR fragment-based PCRs can be used to identify A. pleuropneumoniae from other bacterial swine pathogens and will be useful in surveillance and epidemiological studies.

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