Ali Tevfik UNCU, Mustafa Emin ÖZ, Emrah TORLAK, Müge DOĞAN, Oğuzhan AVCI

A new molecular approach to the diagnosis of small ruminant morbillivirus with EvaGreen based assay

Abstract: The present work describes a new method that is anticipated to contribute to the global control and eradication strategies of small ruminant morbillivirus (SRMV), which negatively affects sheep and goat breeding in many countries where outbreaks emerge. Described assay was developed to detect SRMV by targeting the fusion glycoprotein gene. In the study, the EvaGreen based real-time RT-PCR method is compared with real-time RT-PCR based on TaqMan probe, which is recommended as the diagnostic test by World Organisation for Animal Health (OIE). Detection limit, normalized fluorescence values, and sensitivity of the methods were evaluated in the analysis. The lowest limit of detection obtained was approximately 12 RNA copies per reaction corresponding to a CT value of 36.79. According to the results, it was observed that the developed in-house method has a specific, sensitive, and powerful detection limit as much as the gold standard test. As a result, it was concluded that the proposed method, which was developed as a more costeffective and easily optimized analysis, could provide an advantage for detecting SRMV in diagnostic laboratories.Keywords: EvaGreen, fusion glycoproteingene, low-cost diagnosis, real-time RT-PCR, SRMV

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