title: Simultaneous detection of RNA and DNA targets based on multiplex isothermal amplification.
abstract: The detection of pathogenic microorganisms present in food, feed, plant, and other samples is important for providing safe food as well as for preventing the spread of microbes. The genome of pathogens is made of DNA or RNA, therefore a multiplex diagnostics tool would ideally be able to amplify and detect both RNA and DNA targets in parallel. With this goal we have developed an isothermal nucleic acid sequence based amplification [NASBA] implemented microarray analysis (NAIMA) procedure, suitable for the simultaneous multiplex amplification of RNA and DNA targets, coupled with the detection on ArrayTubes. The method is demonstrated to be very sensitive and specific for the detection of two economically important quarantine plant pathogens of potato, the potato spindle tuber viroid (RNA target) and Ralstonia solanacearum (DNA target). Because of its isothermal amplification and simple detection equipment, the method is also applicable for on-site analyses. NAIMA can be used in any domain where there is the need to detect RNA and DNA targets simultaneously.
authors.affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology , Večna pot 11, Ljubljana 1000, Slovenia.
authors.forename: David
authors.initials: D
authors.lastname: Dobnik
authors.forename: Dany
authors.initials: D
authors.lastname: Morisset
authors.forename: Rok
authors.initials: R
authors.lastname: Lenarčič
authors.forename: Maja
authors.initials: M
authors.lastname: Ravnikar
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000382:microbiology / Q000821:virology; D012367:RNA, Viral / Q000235:genetics*; D043368:Ralstonia solanacearum / Q000235:genetics / Q000302:isolation & purification*; D011198:Solanum tuberosum / Q000382:microbiology* / Q000821:virology*; D014772:Viroids / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial; D012367:RNA, Viral