TITLE:
Effects of Biological Materials and Collection Media on PCR Detection of Tritrichomonas foetus
AUTHORS:
Kristin A. Clothier, Bret Mc Nabb, Andrea Torain, Steve Reinl, Jeff Ondrak
KEYWORDS:
Tritrichomonas foetus, PCR Detection, Urine, Semen, Blood
JOURNAL NAME:
Open Journal of Animal Sciences,
Vol.9 No.1,
January
15,
2019
ABSTRACT:
Tritrichomonas foetus is an important pathogen of the bovine reproductive tract causing early
embryonic death and abortion in cows and persistent, asymptomatic infection in
bulls. PCR detection methods have greatly enhanced diagnostic accuracy over
culture; however, pre-analytical sample handling is just as critical as
technical performance in detecting this pathogen and is not well studied. The
purpose of this study was to investigate the effects of biological materials
present in the prepuce on PCR detection of T.
foetus in a variety of collection media. Simulated preputial samples were created using InPouchTM (IP) media, lactated ringers solution (LRS), or sterile saline (SAL);
inoculated with low numbers of one of three T.
foetus strains; and spiked with either blood, semen, urine, or sham
treatment. Samples were transported to the lab, placed in growth media (LRS and
SAL samples), incubated, and tested for T.
foetus by PCR. Samples containing urine had statistically significantly
greater mean Ct values (P = 0.008)
than samples containing other materials, seen most dramatically in IP (P P = 0.037)
fewer samples being identified as “positive” for T. foetus. Overall, SAL collections also had significantly higher
mean Ct than IP or LRS (P P =
0.018) to results in classification as a “positive” sample. Results of this
study indicate that collection media and biological materials can affect T. foetus PCR detection. The presence of
urine in preputial samples can result in false negative results, while blood
had no detrimental effects.