Usefulness of PCR/RFLP and ERIC PCR techniques for epidemiological study of Haemophilus parasuis infections in pigs

Abstract

Haemophilus parasuis belongs to opportunistic microorganisms of undefined virulence. The purpose of the studies was to compare suitability of PCR/RFLP in our modification and ERIC PCR for epidemiological study of domestic strains of H. parasuis. The results were evaluated taking into account two different aspects: suitability of the tests for isolating the highest possible number of clone groups and subjective evaluation of the method judged with respect to the following criteria: difficulty, availability of equipment and reagents as well as time and cost of the study. The results obtained in the present study show that the two methods used for typing of H. parasuis had high discriminatory power. Taking into account this parameter it can be concluded that ERIC PCR is more suitable than PCR/RFLP. This justifies the use of ERIC PCR for routine epidemiological analyses of mentioned pathogen. Taking into account the complexity of method used, ERIC-PCR based on random amplification of DNA, proved to be comparable to PCR/RFLP. The last mentioned technique is relatively less expensive and labour-consuming, especially when diagnostic PCR method is used for the epidemiological studies.

Comparison of Haemophilus parasuis reference strains and field isolates by using random amplified polymorphic DNA and protein profiles

“…Some H. parasuis isolates were not able to be assayed by using the ERIC-PCR [20] because they gave no or very poor results. Recent studies have found a high diversity of H. parasuis strains isolated in various geographic areas but have not been able to assign a clear correlation between virulence or serovar and ERIC-PCR clusters [19-21]. This conclusion agrees with other H. parasuis ERIC-PCR studies [12,18].…”

“…Although ERIC-PCR has recently been extensively used to study the epidemiology of H. parasuis isolates [19-21], the random amplified polymorphic DNA (RAPD) technique has not been utilized for this purpose. However, RAPD has been used to distinguish other gamma-proteobacteria, including Salmonella spp.…”

Coinfection modulates inflammatory responses, clinical outcome and pathogen load of H1N1 swine influenza virus and Haemophilus parasuis infections in pigs

“…Besides, the above mentioned pathogens, the Haemophilus parasuis (Hps) can also be recovered from the lungs of pigs with pneumonia [1,[7][8][9][10]. In these cases Hps is often isolated along with other bacterial or viral pathogens and, therefore, the role of Hps in producing pneumonia is not clear [8,11].…”

“…It could be also isolated from nasal cavity, tonsils and trachea of apparently healthy pigs [8,14]. Under favorable conditions, Hps can cause severe systemic infection characterized by fibrinous polyserositis, arthritis and meningitis [8,11,14]. Factors leading to systemic infection by Hps have not been clarified to date [9,14].…”

Genotypic analyses and virulence characterization ofGlaesserella parasuisisolates from Taiwan

“…One of the genotyping methods developed to assess the heterogeneity of G. parasuis isolates is enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR), which determines the relationships of G. parasuis isolates by comparing genetic fingerprints (Oliveira, Blackall & Pijoan, 2003; Rafiee et al, 2000). Compared to other genotyping assays, such as restriction fragment length polymorphism, multilocus sequence typing and single-locus sequence typing, ERIC-PCR has higher discriminatory power and lower cost (Jablonski et al, 2011; Olvera, Segales & Aragon, 2007).…”