Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification
The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Hanrimwon Publishing Company
2015-09-01
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| Series: | The Plant Pathology Journal |
| Subjects: | |
| Online Access: | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4564147 |
| _version_ | 1819126541299220480 |
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| author | Joojin Jeong Sang-Yun Cho Wang-Hyu Lee Kui-jae Lee Ho-Jong Ju |
| author_facet | Joojin Jeong Sang-Yun Cho Wang-Hyu Lee Kui-jae Lee Ho-Jong Ju |
| author_sort | Joojin Jeong |
| collection | DOAJ |
| description | The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR. |
| first_indexed | 2024-12-22T07:57:40Z |
| format | Article |
| id | doaj.art-654e4d61cc08416bbb159c7221e56791 |
| institution | Directory Open Access Journal |
| issn | 1598-2254 |
| language | English |
| last_indexed | 2024-12-22T07:57:40Z |
| publishDate | 2015-09-01 |
| publisher | Hanrimwon Publishing Company |
| record_format | Article |
| series | The Plant Pathology Journal |
| spelling | doaj.art-654e4d61cc08416bbb159c7221e567912022-12-21T18:33:19ZengHanrimwon Publishing CompanyThe Plant Pathology Journal1598-22542015-09-0131321922510.5423/PPJ.OA.03.2015.0044PPJ.OA.03.2015.0044Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal AmplificationJoojin Jeong0Sang-Yun Cho1Wang-Hyu Lee2Kui-jae Lee3Ho-Jong Ju4Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, KoreaDepartment of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, KoreaDepartment of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, KoreaDivision of Biotechnology, Chonbuk National University, Iksan 570-752, KoreaDepartment of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, KoreaThe primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4564147loop primer setsPotato virus XRT-LAMP reactionvirus detection |
| spellingShingle | Joojin Jeong Sang-Yun Cho Wang-Hyu Lee Kui-jae Lee Ho-Jong Ju Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification The Plant Pathology Journal loop primer sets Potato virus X RT-LAMP reaction virus detection |
| title | Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification |
| title_full | Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification |
| title_fullStr | Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification |
| title_full_unstemmed | Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification |
| title_short | Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification |
| title_sort | development of a rapid detection method for potato virus x by reverse transcription loop mediated isothermal amplification |
| topic | loop primer sets Potato virus X RT-LAMP reaction virus detection |
| url | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4564147 |
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