Optimization and validation of a real time PCR assay for identification and quantification of trichostrongyle nematodes in goats
Towner, Jennifer Nicole
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Improved diagnostic techniques are needed to rapidly differentiate among common trichostrongyle species of ruminants. Recent studies demonstrate that molecular techniques are more reliable and focus on using quantitative PCR (QPCR) as a means for rapidly differentiating trichostrongyle species. The aim of this study is to optimize a QPCR assay in order to identify and quantify DNA of Haemonchus contortus, Trichostrongylus colubriformis, and Teladorsagia circumcincta based on the ribosomal ITS2 sequences. The PCR assay was used to detect and measure the levels of each species present in goat fecal samples received from throughout the United States. QPCR was sensitive at detecting parasites present in low numbers and at detecting species difficult to identify from coproculture. Culture with L3 identification remains the most accurate means for precise quantification of species present in a fecal sample. This study reports the first attempt using QPCR at quantifying fecal samples containing multiple trichostrongyle species.