Loop-mediated isothermal amplification-based detection of Fusarium graminearum

Molecular biological detection and quantification of fungal DNA targets today relies mainly on the -application of the polymerase chain reaction (PCR). However, this well-recognized technique necessitates the use of highly purified DNA, in a well-equipped lab environment by trained personnel. The method has therefore considerable restrictions when it comes to on-site testing applications for phytopathogenic, mycotoxigenic, and medically relevant fungi and yeasts. As opposed to PCR, molecular biological detection of fungal DNA with isothermal amplification methods is performed at constant temperature. This makes thermal cycling superfluous and enables application of molecular detection assays on site. Moreover, detection of amplification product is done in-tube with no post-amplification manipulations necessary. Here we describe the use of loop-mediated isothermal amplification (LAMP) with indirect in-tube detection of DNA amplification as a rapid and robust method. Detection of F. graminearum in pure cultures and in cereal samples will be described as an example for the high potential which LAMP may hold for future developments in fungal detection assays.