Abstract:To establish and assess different methods for detecting Bacillus anthracis from environmental samples by real-time fluorescence polymerase chain reaction,different methods for purification of nucleic acids from soil and tissues contaminated by B.anthracis vaccine strain were established. TaqMan Primers and probes designed from pagA (pXO1) and rpoB2 genes (chromatosome DNA) of B.anthracis were used to assess these methods by real-time fluorescence polymerase chain reaction. The most effective systems were confirmed by comparing the amplification results. It was found that the method of NaI-glassmilk was the most effective way under these conditions,positive results could be obtained at 1 000 copies/ml or more in crude blood specimens mingled vaccine strain, and same as 25 000 copies or more in one gram soil. The NaI-Glassmilk method and the real-time fluorescence polymerase chain reaction could directly and rapidly detect B.anthracis in contaminated issues or soil.
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