Nucleic Acid-based Techniques for Diagnosis

Abstract

Nucleic acid-based methods have the potential to provide accurate and timely diagnosis of fungal plant pathogens to make better informed decisions relating to disease management and control. Due to their sensitivity, specificity, speed and relative ease-of-use these methods can also be useful tools in the investigation of infection processes, disease progression and epidemiology. This chapter describes the key steps for the use of molecular techniques, beginning with advice and guidance on DNA extraction and disruption of material, describes and discusses conventional PCR covering both PCR development but also practical considerations for the use of PCR-based methods. It describes real-time PCR considering both interpretation and quantification and finally discusses isothermal amplification supported by protocols for the key techniques.