title: Comparison and evaluation of three diagnostic methods for detection of beet curly top virus in sugar beet using different visualizing systems.
abstract: To diminish the time required for some diagnostic assays including polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP; due to mainly DNA extraction step) and also triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) into a minimum level, an innovative immunocapture LAMP (IC-LAMP) and immunocapture PCR (IC-PCR) protocol on the basis of beet curly top virus (BCTV) genome was used and optimized. TAS-ELISA was employed first to validate the existence of the virus. All six IC-LAMP primers (i.e. forward outer primer (F3), backward outer primer (B3), forward inner primer (FIP), backward inner primer (BIP), loop forward (LF) and loop backward (LB)) together with IC-PCR primers were designed on the basis of the replication-associated protein (rep) gene (GenBank accession AF379637.1) of BCTV genome. Also, a novel colorimetric IC-LAMP assay for rapid and easy detection of BCTV was developed here, its potential compared with TAS-ELISA and IC-PCR assays. The method, on the whole, had the following advantages over the two mentioned procedures: (i) fascinatingly, no need of DNA extraction; (ii) no requirement of expensive and sophisticated tools for amplification and detection; (iii) no post-amplification treatment of the amplicons and (iv) a flexible and easy detection approach, which is visually detected by naked eyes using diverse visual dyes.
authors.affiliation: Department of Biotechnology, Research Institute of Physiology and Biotechnology, University of Zanjan, PO Box 45195-313, Zanjan, Iran, mohammadaminalmasi136301@gmail.com.
authors.forename: Mohammad Amin
authors.initials: MA
authors.lastname: Almasi
authors.forename: Seyed Mohammad
authors.initials: SM
authors.lastname: Hosseyni-Dehabadi
authors.forename: Mehdi
authors.initials: M
authors.lastname: Aghapour-ojaghkandi
mesh_terms: D027461:Beta vulgaris / Q000821:virology*; D003116:Color; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D017791:Geminiviridae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*