Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

• Published in: International journal of molecular sciences 2020-11, Vol.21 (22), p.8741
• Main Authors: Edgü, Güven, Freund, Lena Julie, Hartje, Stefanie, Tacke, Eckhard, Hofferbert, Hans-Reinhard, Twyman, Richard M, Noll, Gundula A, Muth, Jost, Prüfer, Dirk
• Format: Article
• Language: English
• Subjects: Colorimetry; Colorimetry - instrumentation; Colorimetry - methods; Commercialization; Crop diseases; Cultivation; Developing countries; Diagnostic systems; Experiments; Gene amplification; Humans; Laboratories; LDCs; Methods; Molecular Diagnostic Techniques - instrumentation; Molecular Diagnostic Techniques - methods; Molecular Typing - instrumentation; Molecular Typing - methods; Necrosis; Nucleic Acid Amplification Techniques - instrumentation; Nucleic Acid Amplification Techniques - methods; Onsite; Optimization; Pathogens; Plant Diseases - virology; Plant Tubers - virology; Plant Viruses; Potatoes; Proteins; Reproducibility of Results; RNA, Viral - genetics; Sensitivity and Specificity; Signs and symptoms; Solanum tuberosum - virology; Tobacco; Tubers; Viruses
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21228741

Potato is an important staple food crop in both developed and developing countries. However, potato plants are susceptible to several economically important viruses that reduce yields by up to 50% and affect tuber quality. One of the major threats is corky ringspot, which is a tuber necrosis caused by tobacco rattle virus (TRV). The appearance of corky ringspot symptoms on tubers prior to commercialization results in ≈ 45% of the tubers being downgraded in quality and value, while ≈ 55% are declared unsaleable. To improve current disease management practices, we have developed simple diagnostic methods for the reliable detection of TRV without RNA purification, involving minimalized sample handling (mini), subsequent improved colorimetric loop-mediated isothermal amplification (LAMP), and final verification by lateral-flow dipstick (LFD) analysis. Having optimized the mini-LAMP-LFD approach for the sensitive and specific detection of TRV, we confirmed the reliability and robustness of this approach by the simultaneous detection of TRV and other harmful viruses in duplex LAMP reactions. Therefore, our new approach offers breeders, producers, and farmers an inexpensive and efficient new platform for disease management in potato breeding and cultivation.