title: Loop-mediated isothermal amplification for the rapid detection of Chrysanthemum chlorotic mottle viroid (CChMVd).
abstract: Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method offering rapid, sensitive, and convenient diagnosis of infectious diseases. Chrysanthemum chlorotic mottle viroid (CChMVd) causes one of the most serious viral diseases in chrysanthemum in Korea. A sensitive LAMP assay was developed for rapidly detecting CChMVd infection. The assay was based on a set of four primers matching the specific region of the CChMVd genome. The CChMVd LAMP primer sets were designed using the sequences from nonsymptomatic and symptomatic CChMVd isolates in Korea. The efficiency and specificity of this method were optimized using Bst DNA polymerase, which allowed for increased viroid detection sensitivity. The reaction was carried out at 65 °C for 90 min, and was improved by adding SYBR Green I dye to the inside of the reaction tube lid prior to amplification. The results indicate that this LAMP method will be useful for chrysanthemum viroid disease monitoring and detecting CChMVd infectious disease.
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Jungan
authors.initials: J
authors.lastname: Park
authors.forename: Yuchul
authors.initials: Y
authors.lastname: Jung
authors.forename: Eui-Joon
authors.initials: EJ
authors.lastname: Kil
authors.forename: Jaedeok
authors.initials: J
authors.lastname: Kim
authors.forename: Dung
authors.initials: D
authors.lastname: Thi Tran
authors.forename: Seung-Kook
authors.initials: SK
authors.lastname: Choi
authors.forename: Ju-Yeon
authors.initials: JY
authors.lastname: Yoon
authors.forename: Won Kyong
authors.initials: WK
authors.lastname: Cho
authors.forename: Sukchan
authors.initials: S
authors.lastname: Lee
mesh_terms: D031202:Chrysanthemum / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D007723:Korea; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors; D014772:Viroids / Q000235:genetics / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers
keywords: Chrysanthemum; Chrysanthemum chlorotic mottle viroid (CChMVd); Detection; Loop-mediated isothermal amplification (LAMP); Viroid