title: Quantitative real-time polymerase chain reaction for detecting Mycoplasma hyosynoviae and Mycoplasma hyorhinis in pen-based oral, tonsillar, and nasal fluids.
abstract: 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.
authors: Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 5011, USA.jgneto84@huskers.unl.edu.
authors: João Carlos
authors: JC
authors: Gomes Neto
authors: Leslie
authors: L
authors: Bower
authors: Barbara Z
authors: BZ
authors: Erickson
authors: Chong
authors: C
authors: Wang
authors: Matthew
authors: M
authors: Raymond
authors: Erin L
authors: EL
authors: Strait
mesh_terms: D000818:Animals; D003955:Diagnostic Tests, Routine / Q000379:methods / Q000662:veterinary*; D005260:Female; D008137:Longitudinal Studies; D009055:Mouth / Q000382:microbiology; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D045790:Mycoplasma hyorhinis / Q000302:isolation & purification*; D045791:Mycoplasma hyosynoviae / Q000302:isolation & purification*; D009666:Nose / Q000382:microbiology; D014066:Palatine Tonsil / Q000382:microbiology; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D015203:Reproducibility of Results; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000382:microbiology
keywords: Mycoplasma hyorhinis; Mycoplasma hyosynoviae; real-time PCR; swine
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