SYBR® Green RT-qPCR for the Universal Detection of Citrus Viroids

Georgios Vidalakis; Jinbo Wang; Tyler Dang; Tavia Rucker; Sohrab Bodaghi; Shi-Hua Tan; Irene Lavagi-Craddock; Alexandra Syed; Gerardo Uribe; Yi Hsieh; Joana Carbajal-Moreno

doi:10.1007/978-1-0716-1464-8_18

SYBR^® Green RT-qPCR for the Universal Detection of Citrus Viroids

Methods Mol Biol. 2022:2316:211-217. doi: 10.1007/978-1-0716-1464-8_18.

Authors

Georgios Vidalakis¹, Jinbo Wang^{2

3}, Tyler Dang⁴, Tavia Rucker⁴, Sohrab Bodaghi⁴, Shi-Hua Tan⁴, Irene Lavagi-Craddock⁴, Alexandra Syed⁴, Gerardo Uribe⁴, Yi Hsieh⁴, Joana Carbajal-Moreno⁴

Affiliations

¹ Department of Microbiology and Plant Pathology, University of California Riverside, Riverside, CA, USA. [email protected].
² Department of Microbiology and Plant Pathology, University of California Riverside, Riverside, CA, USA. [email protected].
³ United States Department of Agriculture-APHIS-Biotechnology Regulatory Service-Biotechnology Risk Assessment Programs, Riverdale, MD, USA. [email protected].
⁴ Department of Microbiology and Plant Pathology, University of California Riverside, Riverside, CA, USA.

PMID: 34845697
DOI: 10.1007/978-1-0716-1464-8_18

Abstract

Quantitative polymerase chain reaction (qPCR) and reverse transcription (RT)-qPCR have now become the gold standard for molecular diagnostics because of its sensitivity, specificity, and reproducibility. In addition, qPCR diagnostics are flexible because they can be scaled for high- or low-throughput applications. Here we describe an optimized assay and workflow for the universal detection of eight citrus viroid species and their variants by RT-qPCR. The assay allows for quick and efficient molecular detection of viroids without the need to run RT-qPCR for each individual viroid species.

Keywords: CCPP; CDFA; Degenerate primers; High-throughput screening; Melting temperature.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Citrus*
Plant Diseases
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Viroids* / genetics