Identification of Pasteurella multocida by polymerase chain reaction

Abstract

Pasteurellosis having a fairly wide distribution can be a reason that hinders the successful development of poultry farming. The causes and conditions for the occurrence of this disease in farms are often not specific. Pasteurella multocida can manifest itself from an extremely weakened Pasteurella carrier to a highly virulent pathogen. A sick bird is a hidden carrier of this disease and is subject to further culling. All this ultimately leads to significant economic losses. Rapid and reliable detection of this pathogen will reduce or completely prevent economic losses. The clinical manifestation of the disease in the form of a lightning course causes difficulties in its lifetime diagnosis. The aim of our work was to develop unique samples of oligonucleotide sequences of primers specific to the pathogen P. multo-cida. The proposed method will provide detection of Pasteurella multocida within 3-4 hours, as well as rapid and highly specific determination of the type of this bacterium. After analyzing the Pasteurella multo-cida genome, we selected a region of the ptfA gene for the construction of oligonucleotides. A pair of primers were selected for PCR: Pm0567 and Pm1321. Gene sequences were aligned by selecting similar ones in absolutely all the studied Pasteurel-la multocida isolates. Using special computer programs, the specificity of the selected primers was evaluated. A search in the sequence database revealed 100% homology of the selected primers with only homologous sequences in the Pasteurella multo-cida genome. The study of samples containing pathologic agents of bacterial nature by PCR confirmed the specificity of the selected primers. Positive results were obtained only with samples containing Pasteurella multocida. Thus, the selected primers can be proposed for the study of samples of different compositions to identify the genome of Pasteurella multocida.