PCR به روش BACILLS ANTRACS تشخيص سريع

Detection of Bacillus Anthracis Via PCR

Routine microbiological methods like biochemical tests are useful for identification of Bacillus Anthracis, but definitive identification may take 24-48 hours. Futher analysis like virulence needs even more time. To establish a rapid and specific detection and identification method for practial applications, the polymerase chain reaction (PCR) was used to identify B. Anthracis using an assay capable of amplifying specific fragments from the gene that encodes the protective antigen (PA), lethal factor (LF), and edema factor (ED) which are Essential for virulence of B. Anthracis. The sequence specific oligonucleotide primers (two PXOI, and PXO2 plasmids and one chromosomally located target) amplified exact fragments only from B. Anthracis (Stern strain) DNA but not related species belonging to the Bacillus Cereus group (including B. Cereus, Bacillus Mycoides and other bacterial strains). This method was able to detect B. Anthracis from extracted DNA and direct inocculation of B. Anthracis cells. Sensitivity analysis by DNA concentration and CFU assay revealed that this method is able to detect less cells in the sample. With optimization of sample preparation, PCR conditions, and fragment analysis (pre PCR, PCR and post PCR) we were able to develop sensitive, accurate, and rapid PCR method for direct detection of B. Anthracis in less than 2 hours with standard thermocyclers.