Molecular Identification and Serogrouping of Pasteurella mutocida Field Isolats

Abstract

Haemorrhagic septicaemia (HS) is a serious disease affecting buffalo and cattle worldwide mainly caused by Pasteurella multocida (PM) serogroup B and E. In Indonesia cases of HS have been reported every year from 2005 to 2017. The bacterial identification was done primarily based on morphological and biochemical characteristics, while the serogroup has never been determined. This research was aimed to implement molecular biological techniques for identification and serogrouping of three archival PM isolates; two isolates were originated from HS cases in South Sulawesi (SulSel) and one isolate from East Nusa Tenggara (NTT). Polymerase chain reaction (PCR) amplifications of PM family-specific gene 16SrRNA, species-specific gene kmt and serotype-specific gene bcbD identified all of the isolates were PM, which was substantiated by inoculation of VITEK® 2 compact. The three PM isolates were belong to capsular serogroup B. Sequence analysis of 690 nucleotides of the capsular gene and the respective hypothetical amino acid was further confirmed their identity. Two isolates of SulSel and NTT were 100% identical to M1404 B: 2 (AF169324.1), meanwhile one silent mutation was noted in the other SulSel isolate. These finding substantiate the application of PCR as a rapid, sensitive and specific method for PM identification and characterization.