Quantitative real-time polymerase chain reaction for detecting Mycoplasma hyosynoviae and Mycoplasma hyorhinis in pen-based oral, tonsillar, and nasal fluids

• Published in: Journal of veterinary science (Suwŏn-si, Korea) Vol. 16; no. 2; pp. 195 - 201
• Main Authors: Gomes Neto, João Carlos, Bower, Leslie, Erickson, Barbara Z, Wang, Chong, Raymond, Matthew, Strait, Erin L
• Format: Journal Article
• Language: English
• Published: Korea (South) 대한수의학회 2015; The Korean Society of Veterinary Science
• Subjects: Animals; Diagnostic Tests, Routine - methods; Diagnostic Tests, Routine - veterinary; Female; Longitudinal Studies; Mouth - microbiology; Mycoplasma hyorhinis - isolation & purification; Mycoplasma hyosynoviae - isolation & purification; Mycoplasma Infections - diagnosis; Mycoplasma Infections - microbiology; Mycoplasma Infections - veterinary; Nose - microbiology; Original; Palatine Tonsil - microbiology; Real-Time Polymerase Chain Reaction - veterinary; Reproducibility of Results; Swine; Swine Diseases - diagnosis; Swine Diseases - microbiology; 수의학; swine; real-time PCR; Mycoplasma hyosynoviae; Mycoplasma hyorhinis
• ISSN: 1229-845X; 1976-555X
• DOI: 10.4142/jvs.2015.16.2.195

Mycoplasma (M.) hyorhinis and M. hyosynoviae are pathogens known to cause disease in pigs post-weaning. Due to their fastidious nature, there is increased need for culture-independent diagnostic platforms to detect these microorganisms. Therefore, this study was performed to develop and optimize quantitative real-time PCR (qPCR) assays to rapidly detect M. hyorhinis and M. hyosynoviae in pen-based oral fluids as well as nasal and tonsillar fluids as proxies for samples used in swine herd surveillance. Two methods of genomic DNA extraction, automated versus manual, were used to compare diagnostic test performance. A wean-to-finish longitudinal study was also carried out to demonstrate the reproducibility of using pen-based oral fluids. Overall, pen-based oral and tonsillar fluids were more likely to be positive for both types of bacteria whereas only M. hyorhinis was detected in nasal fluids. DNA extraction protocols were shown to significantly influence test result. Although the initial detection time somewhat differed, both organisms were repeatedly detected in the longitudinal study. Overall, this study evaluated two qPCR methods for rapid and specific detection of either mycoplasma. Results from the present investigation can serve as a foundation for future studies to determine the prevalence of the two microorganisms, environmental load, and effectiveness of veterinary interventions for infection control.