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Rapid, Simple and Cost-Effective Molecular Method to Differentiate the Temperature Sensitive (ts+) MS-H Vaccine

Zsuzsa Kreizinger1, Kinga Mária Sulyok1, Alexandra Pásztor1

  • 1Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Pest, Hungary.

Plos One
|July 25, 2015
PubMed

Insights

New molecular assays differentiate the temperature-sensitive MS-H vaccine strain from field isolates of Mycoplasma synoviae, crucial for effective poultry vaccination programs. These simple, cost-effective MAMA tests offer high specificity and sensitivity for Mycoplasma synoviae detection.

Area of Science:

  • Veterinary Microbiology
  • Molecular Diagnostics
  • Avian Pathology

Background:

  • Mycoplasma synoviae (M. synoviae) causes significant economic losses in poultry due to respiratory disease and synovitis.
  • Accurate differentiation between the vaccine strain (ts+ MS-H) and field isolates is essential for successful M. synoviae vaccination programs.
  • Existing methods for strain differentiation can be complex or lack sufficient accuracy.

Purpose of the Study:

  • To develop and validate simple, sensitive, and specific molecular assays for differentiating the ts+ MS-H vaccine strain from wild-type M. synoviae isolates.
  • To enable reliable monitoring of M. synoviae vaccination programs.
  • To provide a cost-effective diagnostic tool for avian mycoplasmosis.

Main Methods:

  • Development of melt-curve and agarose gel-based Mismatch Amplification Mutation Assays (MAMA).
  • Targeting single nucleotide polymorphisms (SNPs) at nt367 and nt629 of the M. synoviae obg gene.
  • Validation using clinical samples and comparison with other Mycoplasma species.

Main Results:

  • Two melt-MAMAs and two agarose-MAMAs successfully distinguished the ts+ MS-H vaccine strain from non-temperature-sensitive re-isolates and wild-type M. synoviae.
  • Assays demonstrated high specificity, with no cross-reactions observed with other avian Mycoplasma species.
  • Sensitivity levels of 10^3 (melt-MAMA) and 10^4 (agarose-MAMA) copy numbers were achieved.
  • Assays are adaptable for use in laboratories with basic equipment and can be performed directly on clinical samples.

Conclusions:

  • The developed melt-MAMA and agarose-MAMA assays are reliable tools for differentiating M. synoviae strains.
  • These assays offer simplicity, cost-effectiveness, high sensitivity, and specificity for M. synoviae vaccine strain monitoring.
  • The assays facilitate accurate diagnostics in avian mycoplasmosis control programs.

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