Fluorescent detection techniques for real-time multiplex strand specific detection of Bacillus anthracis using rapid PCR

Speed is a key area in our development of PCR assays for Bacillus anthracis . We believe that the strand specific detection of amplicons within 10 min is a realistic goal and that this will be achieved through fluorescent in-t ube assays. We have used the Idaho LightCycler(TM) to study and develop can didate assays for B. anthracis. New strand specific fluorescent methods hav e been developed and a number of formats have; been studied for speed and s ensitivity. Internal controls have been developed as a method of improving our assay confidence. In this communication we will introduce the held of r apid PCR whilst discussing previous work in the areas described above, the development of our own rapid assay and a novel internal control system for B. anthracis. This work used PCR assays and hardware that are either commer cially available, or have been previously described in open literature publ ications.