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Arbitrarily primed PCR analysis of Mycoplasma hyopneumoniae field isolates demonstrates genetic heterogeneity
1Veterinary Medical Research Institute, Iowa State University, Ames 50011, USA.
Abstract:
Mycoplasma hyopneumoniae is the primary agent of mycoplasmal pneumonia in swine. In this study we performed an arbitrarily primed PCR (AP-PCR) analysis, in which low-stringency amplification with a single primer was used, to investigate genetic variability in M. hyopneumoniae strains and field isolates. We performed preliminary experiments to examine the efficacy of 40 different 10-mer oligonucleotides for priming an AP-PCR with M. hyopneumoniae JT (T = type strain) chromosomal DNA. On the basis of our results, we selected primers OPA-3, OPA-17, and OPB-10 for use in an analysis performed with 23 field isolates. The most informative results were obtained with primer OPA-3. A total of 21 of 23 clinical isolates produced multiband patterns with this primer, while 2 isolates failed to produce any detectable bands. Our data show that M. hyopneumoniae is genetically diverse and that M. hyopneumoniae strains can be divided into at least six epidemiological subgroups on the basis of AP-PCR results.
Insights
Mycoplasma hyopneumoniae, a key swine pathogen, shows significant genetic diversity. Arbitrarily primed PCR (AP-PCR) successfully differentiated field isolates into at least six subgroups, aiding epidemiological studies.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
- Swine Disease Research
Background:
- Mycoplasma hyopneumoniae is the primary cause of mycoplasmal pneumonia in pigs.
- Understanding the genetic diversity of M. hyopneumoniae is crucial for effective disease control and epidemiology.
Purpose of the Study:
- To investigate the genetic variability among Mycoplasma hyopneumoniae strains and field isolates.
- To evaluate the utility of arbitrarily primed PCR (AP-PCR) for differentiating M. hyopneumoniae isolates.
Main Methods:
- Arbitrarily primed PCR (AP-PCR) was employed using single primers for low-stringency amplification.
- Forty different 10-mer oligonucleotide primers were screened for efficacy.
- Primers OPA-3, OPA-17, and OPB-10 were selected for analysis of 23 field isolates.
Main Results:
- Primer OPA-3 yielded the most informative results, producing multiband patterns in 21 out of 23 clinical isolates.
- Two isolates did not produce detectable bands with primer OPA-3.
- The AP-PCR analysis demonstrated significant genetic diversity within M. hyopneumoniae populations.
Conclusions:
- Mycoplasma hyopneumoniae exhibits considerable genetic diversity.
- AP-PCR analysis, particularly with primer OPA-3, can effectively differentiate M. hyopneumoniae strains.
- The results suggest that M. hyopneumoniae strains can be classified into at least six epidemiological subgroups based on AP-PCR profiles.