Development of loop-mediated isothermal amplification LAMP method for rapid detection of Coconut cadang-cadang viroid CCCVd variants in oil palm


Citation

Lau Wei Hong, . and Kong Lih Ling, . and Ganesan Vadamalai, . and Sathis Sri Thanarajoo, . and Jugah Kadir, . (2011) Development of loop-mediated isothermal amplification LAMP method for rapid detection of Coconut cadang-cadang viroid CCCVd variants in oil palm. [Proceedings Paper]

Abstract

Coconut cadang-cadang viroid CCCVd is the causal agent of the lethal Coconut cadang-cadang disease in the Philippines. CCCVd is also associated with orange spotting disease in oil palm. In 2006 Vadamalai and others have demonstrated that oil palms in Malaysia contained several sequence variants of CCCVd. These oil palm CCCVd variants were found in low concentration thus difficult to detect. Rapid detection of CCCVd is vital for prevention and control measures of OS. There are many methods for detecting CCCVd such as Polyacrylamide gel electrophoresis PAGE Ribonuclease protection Assay RPA RT-PCR and hybridization but they are time consuming and inconsistent for CCCVd oil palm variants. Therefore Reverse Transcription Loop-mediated Isothermal Amplification RT-LAMP a novel gene amplification method was evaluated for detection of oil palm CCCVd variants. CCCVd was detected within 60 min by the RT-LAMP method using total nucleic acid extracted from oil palm leaves. This method was able to detect the presence of CCCVd in the infected leaf samples where fluorescence reagent showed colour change from orange to green. Analysis of RT-LAMP product using 2 agarose gel electrophoresis also showed bands of expected size ca 150 bp thus confirming the presence of CCCVd in the oil palm samples.


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Abstract

Coconut cadang-cadang viroid CCCVd is the causal agent of the lethal Coconut cadang-cadang disease in the Philippines. CCCVd is also associated with orange spotting disease in oil palm. In 2006 Vadamalai and others have demonstrated that oil palms in Malaysia contained several sequence variants of CCCVd. These oil palm CCCVd variants were found in low concentration thus difficult to detect. Rapid detection of CCCVd is vital for prevention and control measures of OS. There are many methods for detecting CCCVd such as Polyacrylamide gel electrophoresis PAGE Ribonuclease protection Assay RPA RT-PCR and hybridization but they are time consuming and inconsistent for CCCVd oil palm variants. Therefore Reverse Transcription Loop-mediated Isothermal Amplification RT-LAMP a novel gene amplification method was evaluated for detection of oil palm CCCVd variants. CCCVd was detected within 60 min by the RT-LAMP method using total nucleic acid extracted from oil palm leaves. This method was able to detect the presence of CCCVd in the infected leaf samples where fluorescence reagent showed colour change from orange to green. Analysis of RT-LAMP product using 2 agarose gel electrophoresis also showed bands of expected size ca 150 bp thus confirming the presence of CCCVd in the oil palm samples.

Additional Metadata

[error in script]
Item Type: Proceedings Paper
Additional Information: Available at Perpustakaan Sultan Abdul Samad Universiti Putra Malaysia 43400 UPM Serdang Selangor Malaysia. QR22 M3I61 2011 vol.2 Call Number
AGROVOC Term: Coconut cadang cadang
AGROVOC Term: Viroids
AGROVOC Term: Elaeis guineensis
AGROVOC Term: Oil palms
AGROVOC Term: Leaves
AGROVOC Term: RNA
AGROVOC Term: Nucleic acids
AGROVOC Term: Methodology
AGROVOC Term: Gel electrophoresis
AGROVOC Term: Turbidity
Geographical Term: MALAYSIA
Depositing User: Ms. Suzila Mohamad Kasim
Last Modified: 24 Apr 2025 05:15
URI: http://webagris.upm.edu.my/id/eprint/12987

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