A SYBR green I real-time polymerase chain reaction (PCR) assay for detection and quantification of Trichomonas gallinae
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A SYBR green I real-time polymerase chain reaction (PCR) assay for detection and quantification of Trichomonas gallinae Zaida Rentería-Solís 1,2
&
Tran Nguyen-Ho-Bao 1 & Shahinaz Taha 1 & Arwid Daugschies 1,2
Received: 10 April 2020 / Accepted: 13 September 2020 / Published online: 22 September 2020 # The Author(s) 2020
Abstract Trichomonas gallinae are parasitic flagellates of importance in wild and domestic birds. The parasite is worldwide distributed, and Columbine birds are its main host. Current research focuses mostly on epidemiological and phylogenetic studies. However, there is still a lack of knowledge regarding parasite-host interaction or therapy development. Real-time PCR is a useful tool for diagnostic and quantification of gene copies in a determined sample. By amplification of a 113-bp region of the 18S small subunit ribosomal RNA gene, a SYBR green-based real-time PCR assay was developed. A standard curve was prepared for quantification analysis. Assay efficiency, linearity, and dissociation analysis were successfully performed. Specificity, sensibility, and reproducibility analysis were tested. This assay could be a useful tool not only for diagnostic purposes but also for future in vivo and in vitro T. gallinae studies. Keywords Trichomonas gallinae, . Real-time PCR, . SYBR green, . Trichomoniasis, . Flagellates
Introduction Trichomonas gallinae is a flagellate parasite with a wide number of wild and domestic avian hosts (Forrester and Foster 2008). It infects the upper respiratory and digestive tract. Birds can present mild to fatal disease, also called trichomonosis (Narcisi et al. 1991; Forrester and Foster 2008; Amin et al. 2014). Asymptomatic infections (trichomoniasis) are also common. Trichomonosis is characterised mainly by mild to severe lesions in the oral cavity, throat and crop. Other organs like the liver, lungs and air sacs can also be affected. Severity of the infection is related to strain virulence (Narcisi et al. 1991; Forrester and Foster 2008). Domestic and wild columbiformes are the main host of T. gallinae (Villanúa et al. 2006; Bunbury et al. 2007; Handling Editor: Una Ryan * Zaida Rentería-Solís [email protected]; [email protected] 1
Institute for Parasitology, Centre for Infectious Diseases, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 35, 04103 Leipzig, Germany
2
Albrecht-Daniel-Thaer Institute, An den Tierkliniken 29, 04103 Leipzig, Germany
Forrester and Foster 2008; Amin et al. 2014; Quillfeldt et al. 2018). Birds of prey, particularly Falconiformes are also a major host (Krone et al. 2005; Forrester and Foster 2008; Amin et al. 2014; Quillfeldt et al. 2018). Additionally, T. gallinae has also been reported in other birds such as Psittaciformes and very occasionally, domestic fowl (Amin et al. 2014). Trichomonosis outbreaks can lead to considerable decrease in avian populations (Lawson et al. 2011). Therefore, T. gallinae is considered a parasite of importan
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