alexa Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification
ISSN: 2157-7471

Journal of Plant Pathology & Microbiology
Open Access

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Research Article

Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

Mohammad Amin Almasi1*, Hossein Jafary2, Aboubakr Moradi1, Neda Zand2, Mehdi Aghapour Ojaghkandi1 and Saeedeh Aghaei1

1Department of Plant Biotechnology, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

2Department of Plant Protection, Agricultural and Natural Resources Research Center of Zanjan, Zanjan, Iran

*Corresponding Author:
Mohammad Amin Almasi
Department of Plant Biotechnology
Faculty of Agriculture
University of Zanjan
Zanjan, Iran
E-mail: [email protected]

Received date: November 19, 2012; Accepted date: December 29, 2012; Published date: January 03, 2013

Citation: Almasi MA, Jafary H, Moradi A, Zand N, Ojaghkandi MA, et al. (2013) Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification. J Plant Pathol Microb 4:156. doi:10.4172/2157-7471.1000156

Copyright: © 2013 Almasi MA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Loop-mediated isothermal amplification assay amplifies DNA/RNA with high specificity and sensitivity. In this study, we describe an optimized reverse transcription- LAMP assay for detection of Potato Leafroll Virus. Firstly, DAS-ELISA assay was performed to detect of the virus in a collection containing 40 suspicious samples. Lastly, two samples were detected as the positive samples. Then, the positive samples were verified by RT-PCR and RT-LAMP methods. Furthermore, the results demonstrated that the RT-LAMP assay was 40 times sensitive and 4 time faster compared to RT-PCR. RT-LAMP assay was accomplished in the water bath either frees from any thermal cycler machine or sophisticated laboratories facility. Moreover, in RT-LAMP reaction the positive samples were detected through turbidity which produced by magnesium pyrophosphate. Interestingly, the application of CaCl2 instead of
MgSO4 which create calcium pyrophosphate in reaction could significantly increase both stability and concentration of turbidity. Consequently, it could be an interesting alternative to MgSO4. Overall, the newly developed RT-LAMP assay can be a sensitive, specific and low-cost method for early detection of Potato Leafroll Virus and also other viral plant pathogens.

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