Multiplex Polymerase Chain Reaction Assay for the Specific Detection of the Organism Causing Anthrax

The zoonotic nature and potential to be used as an agent of bioterrorism, quick onset and rapid lethal progression of the disease, ability to persist in environment for decades etc. necessitates the development of specific, sensitive, safe and reliable detection methods for the organism causing anthrax. Therefore, an attempt has been made in the present study to optimize a multiplex polymerase chain reaction (PCR) for the specific and sensitive identification of bacteria causing anthrax. Three sets of primers, one from chromosome (bxpB) and two from pXO1 plasmid (pag and cya) were designed so as to detect entire Bacillus cereus group and then specifically the bacterium causing anthrax, by single reaction. The selected primers amplified the desired products from both Sterne strain and a field isolate of Bacillus anthracis. The PCR amplification with template of other bacteria of same and different genera yielded negative results. The sensitivity was 1 pg of total DNA. The present article also delineates a very simple, reliable and less risky method for the extraction of both plasmid and chromosomal DNA from B. anthracis.