Real-time polymerase chain reaction for the qualitative and quantitative detection of Mycoplasma gallisepticum.
The aim of this study was to evaluate the sensitivity and the detection limit of a real-time polymerase chain reaction (Q-PCR) developed for the qualitative and quantitative detection of Mycoplasma gallisepticum. No cross-reactivity was observed with DNA from other important avian mycoplasmas, including Mycoplasma synoviae and Mycoplasma meleagridis. However, the Q-PCR could not distinguish between M. gallisepticum and Mycoplasma imitans. The Q-PCR had detection limits 10 to 1000 times lower than a conventional commercial PCR method and than culture. The Q-PCR was used quantitatively by incorporating a set of external M. gallisepticum DNA standards, derived from a M. gallisepticum log-phase culture of a known concentration. The number of colony-forming unit equivalents per millilitre in tracheal swabs from experimentally infected birds could be determined from a single sample. The method had good reproducibility and correlated well with standard counting techniques using culture. It can be concluded that the Q-PCR described is suitable for qualitative and quantitative detection of M. gallisepticum in clinical samples.
Mekkes
D R
DR
Animal Health Service, PO Box 9, 7400 AA, Deventer, The Netherlands.
Feberwee
A
A
D000818:Animals; D002645:Chickens / Q000382:microbiology*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D045686:Mycoplasma gallisepticum / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology; D012680:Sensitivity and Specificity; D013045:Species Specificity