Molecular characterization of the capsular antigens of Pasteurella multocida isolates using multiplex PCR

Abstract

The use of molecular techniques for detection and characterization of the Pasteurella multocida is very important for rapid and specific detection and characterization of the organism. During the period from 15th February, 2014 to 15th April, 2015, 425 nasopharyngeal swabs and 175 lung and spleen samples were collected and examined by conventional methods, 80 strains (18.82%) of P. multocida were isolated from the calves, sheep and goat with respiratory manifestation. Meanwhile, 77 strains (44%) were isolated from emergency slaughtered animals. All the recovered strains were positive for specific PCR for detection of P. multocida strains previously identified as P. multocida by standard microbiological techniques. Multiplex PCR for molecular typing of the capsular antigens of the recovered P. multocida revealed positive amplification of 1044 bp fragments specific to the capsular antigen type A with 105 strains (66.88%), and amplification 511 bp fragments of the capsular antigen type E with 52 strain (33.12%) and absence of B, D and F antigens. Multiplex PCR for molecular typing of the capsular antigens of P. multocida can be used as a simple, sensitive, rapid, reliable technique instead of the serological techniques for identification of the capsular antigens of P. multocida.

Keywords: Capsular antigens; Microbiological techniques; Molecular typing; Multiplex PCR; P. multocida.

Figure 1

Agarose gel electrophoresis showing amplification of 460 bp fragments specific for P. multocida field isolates.

Figure 2

Agarose gel electrophoresis showing amplification of 1044 bp fragments specific to the capsular antigen type A and 511 bp fragments specific to the capsular antigen type E.