Development of conventional and taqman real-time polymerase chain reaction methods for detection of liver fluke infection with platynosomum spp. In cat feces

Abstract

The Platynosomum spp. fluke is the etiologic agent of platynosomiasis responsible for hepatobiliary diseases in cats. Coprological examination using centrifugal sedimentation technique is the gold standard method for detection of Platynosomum spp.’s eggs. However, microscopic examination method has a limited sensitivity, particularly in cases of complete biliary obstruction and inconsistent egg shedding. To circumvent these problems, this study aims to develop molecular-based methods for detecting Platynosomum spp. infection in cat feces including conventional PCR (cPCR) and TaqMan real-time PCR methods targeting ITS1 region. From a total number of 120 cat fecal samples, when compared to microscopic examination, the cPCR method had sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 80.0%, 88.3%, 87.3%, and 81.5%, respectively, while the TaqMan real-time PCR method had these values of 98.3%, 88.3%, 89.4%, and 98.2%, respectively. Furthermore, the TaqMan real-time PCR can assess a greater prevalence (55%; 66/120) than that of the gold standard microscopic (50%; 60/120) and cPCR (45.8%; 55/120) methods. The TaqMan real-time PCR method developed in this study showed high and acceptable values of these parameters suggesting that this technique could be used to increase the detect rate of Platynosomum spp. infection from the cats with false negative results by gold standard method.