Application of Polymerase Chain Reaction Fingerprinting to Differentiate Ornithobacterium Rhinotracheale Isolates

Abstract

Ornithobacterium rhinotracheale (ORT) is an infectious respiratory pathogen of chickens, turkeys, and wild birds. There are 18 serotypes of ORT reported worldwide. In this study, enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction and random amplified polymorphic DNA assay with Universal M13 primer-based fingerprinting techniques were investigated for their ability to differentiate ORT isolates. The authors examined 50 field isolates and 8 reference strains of ORT for their genetic differences. The fingerprint patterns were compared with serotyping results of ORT by the agar gel precipitation test. M13 fingerprinting revealed different patterns for 6 reference serotypes of ORT that were tested, namely, C, D, E, I, J, and K. Ornithobacterium rhinotracheale reference serotypes A and F yielded indistinguishable fingerprints with M13 fingerprinting. The ERIC 1R technique discerned only 5 of the 8 reference serotypes of ORT. Distinct fingerprints were also found within the ORT serotypes with both techniques. From 58 isolates of ORT that were fingerprinted belonging to 8 ORT serotypes, 10 different fingerprints were obtained with M13 fingerprinting and 6 different fingerprints were obtained with ERIC 1R fingerprinting. M13 fingerprinting technique was found to be more discriminative in differentiating ORT isolates than the ERIC 1R fingerprinting technique. These results suggest that fingerprinting techniques may be a more discerning tool for characterizing ORT isolates than the serological test using the agar gel precipitation test. This fingerprinting technique could potentially be a valuable tool in identifying an isolate from a clinical outbreak of ORT infection for development of an autogenous vaccine.

Comparison and Utilization of Repetitive-Element PCR Techniques for Typing Lactobacillus Isolates from the Chicken Gastrointestinal Tract

“…The current research extends the application of ERIC-PCR, which has previously been used for typing a wide variety of gram-negative bacteria in chickens, including Arcobacter spp. (5), Escherichia coli (17), Salmonella enterica (13), Campylobacter jejuni (70), Pasteurella multocida (52), and Ornithobacterium rhinotracheale (60). Collectively, the current research and these past studies highlight the utility of ERIC-PCR in the typing of a variety of Isolates of enterococci were also able to be typed to the species and ERIC type levels, supporting a previous assessment of the ERIC-PCR for the differentiation of E. faecium isolates (18).…”

Sero-prevalence and identification of Ornithobacterium rhinotracheale in broiler flocks in south-eastern Iran

“…ORT is an emerging poultry pathogen associated with respiratory disease (Murthy et al 2008). There are 18 serotypes for ORT, with the majority of chicken isolates belonging to four serotypes A, B, D and E (Chansiripornchai et al 2007;Thachil et al 2007). Haemagglutinating activity of ORT serovars has been studied by using fresh erythrocytes from different species (Vega et al 2008).…”

Isolation, identification, antibiotic resistance profile and molecular analysis of Ornithobacterium rhinotracheal isolates from turkeys

“…They reported that the M13 fingerprinting technique was found to be more discriminative in differentiating ORT isolates than the ERIC fingerprinting technique. They suggest that fingerprinting techniques may be a more discerning tool for characterizing ORT isolates (Thachil et al., 2007 ). In this research, the isolates showed 20 different genetic patterns based on ERIC PCR results and were classified into four different clusters.…”