Abstract
Surface plasmon resonance (SPR)-based DNA biosensors have been shown to be rapid, label-free, and selective tools for the detection of PCR products. Here, we describe an SPR sensor based on DNA hybridization for the detection of Fusarium culmorum, a fungal pathogen of wheat. A 0.57 kb DNA fragment of F. culmorum was amplified by specific primers, and a 25-mer oligonucleotide probe was selected within the sequence of the PCR amplicon. The biotin-labeled probe was immobilized on a streptavidin sensor chip and tested for biospecific interaction with PCR products of F. culmorum. The SPR biosensor was applied to the detection of F. culmorum in fungal cultures and in naturally infected wheat samples.
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References
Nirenberg HI (1981) A simplified method for identifying Fusarium spp. occurring on wheat. Can J Botany 59:1599–1609
Edwards SG, Pirgozliev SR, Hare MC, Jenkinson P (2001) Quantification of trichothecene-producing Fusarium species in harvested grain by competitive PCR to determine efficacies of fungicides against fusarium head blight of winter wheat. Appl Environ Microbiol 67:1575–1580
Yli-Mattila T, Paavanen-Huhtala S, Jestoi M, Parikka P, Hietanieme V, Gagkaeva T, Sarlin T, Haikara A, Laaksonen S, Rizzo A (2008) Real-time PCR detection and quantification of Fusarium poae, F. graminearum, F. sporotrichioides and F. langsethiae in cereal grains in Finland and Russia. Arch Phytopathol Plant Protection 41(4):243–260
Brunner K, Mach RL (2010) Quantitative detection of fungi by molecular methods: a case study on Fusarium. In: Voigt K, Gherbawy Y (eds) Molecular identification of fungi. Springer, Berlin, Heidelberg, pp 93–106
Homola J (2008) Surface plasmon resonance sensors for detection of chemical and biological species. Chem Rev 108:462–493
Homola J (2006) Surface plasmon resonance based sensors. Springer, Berlin, Heidelberg, p 251
Zezza F, Pascale M, Mulè G, Visconti A (2006) Detection of Fusarium culmorum in wheat by a surface plasmon resonance-based DNA sensor. J Microbiol Methods 66:529–537
Skottrup P, Hearty S, Frøkiær H, Leonard P, Hejgaard J, O’Kennedy R, Nicolaisen M, Justesen AF (2007) Detection of fungal spores using a generic surface plasmon resonance immunoassay. Biosens Bioelectron 22:2724–2729
Manera MG, Spadavecchia J, Leone A, Quaranta F, Rella R, Dell’Atti D, Minunni M, Mascini M, Siciliano P (2008) Surface plasmon resonance imaging technique for nucleic acid detection. Sens Actuators B Chem 130:82–87
Wagachaa JM, Muthomib JW (2007) Fusarium culmorum: infection process, mechanisms of mycotoxin production and their role in pathogenesis in wheat. Crop Prot 26:877–885
Parry DW, Jenkinson P, McLeod L (1995) Fusarium ear blight (scab) in small grain cereals-a review. Plant Pathol 44:207–238
Logrieco A, Bottalico A, Mulé G, Moretti A, Perrone G (2003) Epidemiology of toxigenic fungi and their associated mycotoxins for some Mediterranean crops. Eur J Plant Pathol 109:645–667
Nicholson P, Simpson DR, Weston G, Rezanoor HN, Lees AK, Parry DW, Joyce D (1998) Detection and quantification of Fusarium culmorum and Fusarium graminearum in cereals using PCR assays. Physiol Mol Plant Pathol 53:17–37
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York
Wilson A, Simpson D, Chandler E, Jennings P, Nicholson P (2004) Development of PCR assays for the detection and differentiation of Fusarium sporotrichioides and Fusarium langsethiae. FEMS Microbiol Lett 233:69–76
Acknowledgments
This work was supported by the Italian Ministry of Education, University and Research (MIUR), Project no. 12792 “SINSIAF”.
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Pascale, M., Zezza, F., Perrone, G. (2013). Surface Plasmon Resonance Genosensor for the Detection of Fusarium culmorum . In: O'Connor, L., Glynn, B. (eds) Fungal Diagnostics. Methods in Molecular Biology, vol 968. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-257-5_12
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DOI: https://doi.org/10.1007/978-1-62703-257-5_12
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