Efficiency of a fluorescent, non-extraction LAMP DNA amplification method: toward a field-based specific detection of maize pollen grains

Loop-mediated isothermal amplification (LAMP) is rapidly being established as the preferred method for field-based, rapid and cost-effective detection of DNA sequences, but a deficiency of current methods for diagnostic purposes is the prevalence of non-specific amplification. We have previously demonstrated the applicability of the LAMP method for pollen from a variety of plant species with a resolution down to a single grain, based on the application of an extraction-free methodology. Here, we show how this method can be applied to detect pollen with sequence-specific accuracy and without the danger of false positives. We establish the lower limits of detection and efficiency of this approach, and in addition, we show that this method does not lose robustness for pollen material stored for at least 8 years.