Rapid and visible detection of Mycoplasma synoviae using a novel polymerase spiral reaction assay

Poult Sci. 2019 Nov 1;98(11):5355-5360. doi: 10.3382/ps/pez356.

Abstract

In this study, a rapid, specific, and sensitive detection assay for Mycoplasma synoviae (MS) was established using a polymerase spiral reaction (PSR) method. A pair of primers were designed according to the conserved region of the vlhA gene of MS, and PSR results were assessed using agarose gel electrophoresis and color rendering with a dye indicator. The optimum reaction temperature and time for PSR using the specific primers were 62°C and 40 min in a water bath, respectively. The sensitivity of the PSR assay for MS detection was 100 times more than that of the polymerase chain reaction assay based on agarose gel electrophoresis results and color change detected by the naked eye. Further experiments demonstrated that the primers specifically detected MS and showed no cross-reaction with other prevalent avian pathogens. Clinical sample testing confirmed that the MS-PSR assay is simple, rapid, specific, and sensitive, and thereby very suitable for application and promotion in the field and laboratories of grassroots units.

Keywords: Mycoplasma synoviae; molecular detection; polymerase spiral reaction.

MeSH terms

  • Animals
  • Chickens*
  • DNA Primers / genetics
  • Mycoplasma Infections / diagnosis
  • Mycoplasma Infections / veterinary*
  • Mycoplasma synoviae / isolation & purification*
  • Nucleic Acid Amplification Techniques / methods*
  • Poultry Diseases / diagnosis*
  • Sensitivity and Specificity

Substances

  • DNA Primers