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Fluorescent detection techniques for real‐time multiplex strand specific detection of Bacillus anthracis using rapid PCR
Author(s) -
Lee M. A.,
Brightwell G.,
Leslie D.,
Bird H.,
Hamilton A.
Publication year - 1999
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.1999.00908.x
Subject(s) - bacillus anthracis , amplicon , multiplex , multiplex polymerase chain reaction , biology , fluorescence , computational biology , real time polymerase chain reaction , microbiology and biotechnology , polymerase chain reaction , bioinformatics , bacteria , gene , genetics , physics , quantum mechanics
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‐tube assays. We have used the Idaho LightCycler TM to study and develop candidate assays for B. anthracis . New strand specific fluorescent methods have been developed and a number of formats have been studied for speed and sensitivity. Internal controls have been developed as a method of improving our assay confidence. In this communication we will introduce the field of rapid 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 commercially available, or have been previously described in open literature publications.