Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP)

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) detected Coconut cadang-cadang viroid (CCCVd) within 60. min at 60. °C in total nucleic acid extracted from oil palm leaves infected with CCCVd. Positive reactions showed colour change from orange to green in the reaction mix a...

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Main Authors: Thanarajoo, Sathis Sri, Kong, Lih Ling, Kadir, Jugah, Lau, Wei Hong, Vadamalai, Ganesan
Format: Article
Language:English
Published: Elsevier 2014
Online Access:http://psasir.upm.edu.my/id/eprint/37013/
http://psasir.upm.edu.my/id/eprint/37013/1/Detection%20of%20Coconut%20cadang-cadang%20viroid%20%28CCCVd%29%20in%20oil%20palm%20by%20reverse%20transcription%20loop-mediated%20isothermal%20amplification%20%28RT-LAMP%29.pdf
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author Thanarajoo, Sathis Sri
Kong, Lih Ling
Kadir, Jugah
Lau, Wei Hong
Vadamalai, Ganesan
author_facet Thanarajoo, Sathis Sri
Kong, Lih Ling
Kadir, Jugah
Lau, Wei Hong
Vadamalai, Ganesan
author_sort Thanarajoo, Sathis Sri
building UPM Institutional Repository
collection Online Access
description A reverse transcription loop-mediated isothermal amplification (RT-LAMP) detected Coconut cadang-cadang viroid (CCCVd) within 60. min at 60. °C in total nucleic acid extracted from oil palm leaves infected with CCCVd. Positive reactions showed colour change from orange to green in the reaction mix after the addition of fluorescent reagent, and a laddering pattern band on 2% agarose gel electrophoresis. Conventional RT-PCR with LAMP primers produced amplicons with a sequence identical to the 297-nt CCCVd oil palm variant with the primers being specific for CCCVd and not for other viroids such as PSTVd and CEVd. RT-LAMP was found to be rapid and specific for detecting oil palm CCCVd.
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institution Universiti Putra Malaysia
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language English
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publishDate 2014
publisher Elsevier
recordtype eprints
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spelling upm-370132015-12-01T02:26:06Z http://psasir.upm.edu.my/id/eprint/37013/ Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP) Thanarajoo, Sathis Sri Kong, Lih Ling Kadir, Jugah Lau, Wei Hong Vadamalai, Ganesan A reverse transcription loop-mediated isothermal amplification (RT-LAMP) detected Coconut cadang-cadang viroid (CCCVd) within 60. min at 60. °C in total nucleic acid extracted from oil palm leaves infected with CCCVd. Positive reactions showed colour change from orange to green in the reaction mix after the addition of fluorescent reagent, and a laddering pattern band on 2% agarose gel electrophoresis. Conventional RT-PCR with LAMP primers produced amplicons with a sequence identical to the 297-nt CCCVd oil palm variant with the primers being specific for CCCVd and not for other viroids such as PSTVd and CEVd. RT-LAMP was found to be rapid and specific for detecting oil palm CCCVd. Elsevier 2014-06-15 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/37013/1/Detection%20of%20Coconut%20cadang-cadang%20viroid%20%28CCCVd%29%20in%20oil%20palm%20by%20reverse%20transcription%20loop-mediated%20isothermal%20amplification%20%28RT-LAMP%29.pdf Thanarajoo, Sathis Sri and Kong, Lih Ling and Kadir, Jugah and Lau, Wei Hong and Vadamalai, Ganesan (2014) Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP). Journal of Virological Methods, 202. pp. 19-23. ISSN 0166-0934; ESSN: 1879-0984 10.1016/j.jviromet.2014.02.024
spellingShingle Thanarajoo, Sathis Sri
Kong, Lih Ling
Kadir, Jugah
Lau, Wei Hong
Vadamalai, Ganesan
Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_full Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_fullStr Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_full_unstemmed Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_short Detection of Coconut cadang-cadang viroid (CCCVd) in oil palm by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_sort detection of coconut cadang-cadang viroid (cccvd) in oil palm by reverse transcription loop-mediated isothermal amplification (rt-lamp)
url http://psasir.upm.edu.my/id/eprint/37013/
http://psasir.upm.edu.my/id/eprint/37013/
http://psasir.upm.edu.my/id/eprint/37013/1/Detection%20of%20Coconut%20cadang-cadang%20viroid%20%28CCCVd%29%20in%20oil%20palm%20by%20reverse%20transcription%20loop-mediated%20isothermal%20amplification%20%28RT-LAMP%29.pdf