Adaptation of a filter paper method for RNA template preparation for the detection of avocado sunblotch viroid by reverse transcription qPCR

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Pretorius, L.-S., Chandra, K. A., Jooste, A. E. C., Motaung, L. C., Parkinson, L. E. and Geering, A. D. W. (2022) Adaptation of a filter paper method for RNA template preparation for the detection of avocado sunblotch viroid by reverse transcription qPCR. Journal of Virological Methods, 301 . p. 114455. ISSN 0166-0934

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Article Link: https://doi.org/10.1016/j.jviromet.2022.114455

Publisher URL: https://www.sciencedirect.com/science/article/pii/S0166093422000027

Abstract

An easy, rapid and inexpensive method of preparing RNA template for a reverse transcription qPCR assay for avocado sunblotch viroid (ASBVd) is described. This method depends on the principle of reversible binding of viroid RNA to filter paper under different concentrations of monovalent cation. Lysis buffers containing either sodium or lithium chloride were compared, and 1.5 M lithium chloride was shown to be optimal for the adsorption of the viroid RNA to the filter paper. The extraction method was validated using field samples and equivalent yields of viroid RNA were obtained using this method and either a commercial RNA extraction kit or a dsRNA chromatography method. The filter paper method of RNA extraction is ideally suited for the large-scale surveillance for ASBVd.

Item Type:	Article
Business groups:	Animal Science
Keywords:	diagnosis surveys RNA extraction
Subjects:	Plant culture > Tree crops Plant culture > Fruit and fruit culture Plant pests and diseases Plant pests and diseases > Plant pathology
Live Archive:	10 Jan 2022 05:13
Last Modified:	07 Jul 2022 00:24

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