Development of a multiplex PCR assay for polymorphism analysis of Brucella suis biovars causing brucellosis in swine

Veterinary Microbiology
Lénia Ferrão-BeckMaria Inácia Sá

Abstract

Swine brucellosis is caused by the biovars 1, 2 and 3 of Brucella suis the identification of which up to now relies on microbiological tests lacking adequate specificity together with time consuming and expensive molecular procedures. Based on sequence variation of the omp2b gene, we have developed a four primer set multiplex PCR assay that was tested for polymorphism analysis of B. suis biovars causing brucellosis in swine. The assay exploits the single nucleotide polymorphisms found in omp2b gene of B. suis reference biovars which are conserved in 43 B. suis field isolates from different geographic origins and hosts. Three specific amplification patterns (S1, S2 and S3) were obtained for reference strains of B. suis biovars 1, 2 and 3, respectively. However, some B. suis field isolates identified as biovars 2 or 3 according AMOS-PCR, PCR-RFLP of omp31 and omp2 genes and classical bacteriological methods, resulted also in S1 patterns, limiting the typing usefulness of the method.

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Citations

Mar 9, 2010·BMC Infectious Diseases·Pilar M MuñozChristian Gortázar
Aug 19, 2010·Croatian Medical Journal·Silvio SpicićZeljko Cvetnić
May 28, 2011·Archives of Virology·Daoliang LanXiuguo Hua
Jan 16, 2016·Journal of Microbiological Methods·Jenni Whiteduck-LéveilléeIzhar U H Khan
Oct 11, 2011·Diagnostic Microbiology and Infectious Disease·Gernot SchmoockSascha Al Dahouk
Jun 30, 2009·International Journal of Medical Microbiology : IJMM·Birgit HuberHans-Jürgen Busse
May 24, 2008·Veterinary Microbiology·David FretinPierre Wattiau
Sep 14, 2017·BMC Genomics·Ana Cristina FerreiraRicardo Dias
Jan 26, 2011·The Veterinary Record·F Closa-SebastiàI Marco

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