Evaluation of nucleic acids extraction methods for effective detection of citrus tristeza virus by TaqMan real‐time RT ‐ PCR

Citrus tristeza virus (CTV) is the primary viral disease impacting citrus worldwide. Given the limitations of available control methods, early detection of CTV is of paramount importance. Several molecular techniques are utilized for virus detection, with TaqMan real‐time RT‐PCR recognized as among the most sensitive ones. Nonetheless, effective CTV detection requires robust RNA extraction methods. In this study, a comparative analysis of RNA yield and purity, real‐time RT‐PCR performance, processing time, and cost of two extraction methods (Semi‐automated magnetic bead‐based method, Silica capture method), and a direct method (Tissue‐print) was undertaken. The Semi‐automated magnetic bead‐based method demonstrated high efficiency with high RNA yield and purity, and high efficiency in terms of real‐time RT‐PCR performance, albeit at a relatively higher cost. The Silica capture method showed moderate RNA yield and purity, real‐time RT‐PCR performance, and cost, but it is more time‐consuming. Tissue‐print was rapid to execute and inexpensive. However, it comes with reduced real‐time RT‐PCR performance and low RNA yield. The results of this study may assist researchers in making informed decisions regarding the selection of an extraction method that aligns with their specific requirements, considering the evaluated criteria.