One-step reverse transcription loop-mediated isothermal amplification for the detection of Capsicum chlorosis virus in Phalaenopsis orchids

Cheng-Ping Kuan; Yu-Fei Lin; Yu-Chun Lin; Chin-Chih Chen

doi:10.1007/s10327-016-0685-4

One-step reverse transcription loop-mediated isothermal amplification for the detection of Capsicum chlorosis virus in Phalaenopsis orchids

Journal

Journal of General Plant Pathology

Journal Volume

82

Journal Issue

6

Pages

307-313

Date Issued

2016-11

Date

2016-11

Author(s)

Cheng-Ping Kuan

Yu-Fei Lin

Yu-Chun Lin

Chin-Chih Chen

DOI

10.1007/s10327-016-0685-4

URI

https://scholars.tari.gov.tw/handle/123456789/4162

Abstract

A one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of Capsicum chlorosis virus (CaCV) was developed and optimized. The RT-LAMP assay was completed within 60 min using isothermal conditions and four primers targeting the CaCV nucleocapsid protein gene. The sensitivity of this assay was higher than that of a reverse transcription polymerase chain reaction using total RNA of infected Phalaenopsis sp. The assay was highly specific allowing CaCV to be differentiated from other orchid-infecting viruses. The result of the assay can be visualized under UV with the addition of SYBR Green I dye to the end product directly in the tube. This assay is a useful tool for the detection of CaCV infected orchids due to its rapidity, simplicity, accuracy and sensitivity.

Subjects

Capsicum chlorosis virus

Detection

Loopmediated isothermal amplification

RT-PCR

Publisher

Springer Nature

Type

journal article

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