Detection of Micro-organisms with the Fluoroskan Ascent® using Molecular Beacons, Taqman®, and SYBR® Green assays

Introduction

Fluorescent labeled probe utilisation has increased in diagnostic laboratories in recent years; uses include medical, agricultural and forestry micro-organism detection and identification. However, several operational diagnostic laboratories do not have access to high-end equipment used in research labs, such as real-time PCR. Here we describe the use of fluorescence detection in endpoint to allow detection and identification of micro-organisms using molecular beacons (3, 4, 5, 6, 7), Taqman (5), or SYBR Green (4).

In this assay, we used specific primers to amplify Phytophthora ramorum, the causal agent of Sudden Oak Death (2,8), combined with fluorescent probes (molecular beacons and Taqman) or non-specific label (SYBR Green). The reactions were analysed in endpoint with Thermo Fluoroskan Accent using endpoint measurement (Figure 1).