The influence of temperature and simulated transport conditions of diagnostic samples on real-time polymerase chain reaction for the detection of Tritrichomonas foetus DNA

Abstract

Bovine trichomoniasis is a sexually transmitted disease in cattle that causes considerable economic loss due to abortions and infertility. In vitro culture of the organisms is the traditional method for diagnosis. However, culture cannot differentiate Tritrichomonas foetus from other, closely related nonpathogenic protozoa. Recently, a quantitative real-time polymerase chain reaction (qPCR) was developed for the differential diagnosis of trichomoniasis. The objective of the current work was to evaluate the effect of different simulated transport conditions on samples containing T. foetus for the diagnosis of trichomoniasis using culture and qPCR. Results indicate that transport temperatures of 4–20°C for 1–3 days before culture will reduce or temporarily inhibit parasite replication but maintain viability. Testing of samples by either culture or qPCR would be expected to give positive results. However, diagnosis of trichomonads by both methods was negatively affected when specimens were maintained at transport temperatures of 42°C for 24 hr or more. The current study stresses the importance of ensuring that clinical samples arrive to the diagnostic laboratory within 24–48 hr and of avoiding temperature transport conditions above 37°C in order to achieve an accurate diagnosis of trichomoniasis in cattle.