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Published on: March 17, 2016
Usefulness of PCR/RFLP and ERIC PCR techniques for epidemiological study of Haemophilus parasuis infections in pigs
A Jabłoński1, S Zebek, R Kołacz
1National Veterinary Research Institute, Swine Diseases Department, 57 Partyzantow Str., 24-100 Pulawy, Poland. artur.jablonski@piwet.pulawy.pl
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.
Insights
ERIC PCR is more suitable for routine epidemiological analysis of Haemophilus parasuis (H. parasuis) than PCR/RFLP due to its high discriminatory power. Both methods offer comparable complexity, but ERIC PCR is recommended for H. parasuis strain typing.
Area of Science:
- Veterinary Microbiology
- Molecular Epidemiology
Background:
- Haemophilus parasuis is an opportunistic pathogen with undefined virulence.
- Accurate strain typing is crucial for epidemiological studies of H. parasuis.
Purpose of the Study:
- To compare the suitability of a modified PCR/RFLP and ERIC PCR for epidemiological studies of domestic H. parasuis strains.
- To evaluate methods based on discriminatory power, ease of use, equipment availability, time, and cost.
Main Methods:
- Polymerase Chain Reaction/Restriction Fragment Length Polymorphism (PCR/RFLP) using a modified protocol.
- Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR.
- Evaluation of discriminatory power and subjective criteria (difficulty, cost, time).
Main Results:
- Both PCR/RFLP and ERIC PCR demonstrated high discriminatory power for H. parasuis typing.
- ERIC PCR showed higher suitability for isolating a greater number of clone groups.
- ERIC PCR is recommended for routine epidemiological analyses of H. parasuis.
Conclusions:
- ERIC PCR is a highly suitable method for routine epidemiological studies of H. parasuis.
- While comparable in complexity, ERIC PCR offers superior discriminatory power for strain differentiation.
- PCR/RFLP remains a viable, less expensive option, particularly for diagnostic PCR applications.
