Detection of spores of Bacillus anthracis from environment using polymerase chain reaction.
A sensitive PCR based detection of Bacillus anthracis spores from environnment was standardized. Specific 1247bp amplicon could be detected with template concentration as low as 13 pg. Sensitivity was enhanced to 10 fold by nesting with second set of primers, forming 208bp amplicon. Extraction of DNA from spores purified from soil samples by aqueous polymer two-phase system followed by partial germination and freeze-thaw treatment yielded best results. Soil sample spiked with spores (8x10(2)/g of sample) could be detected with this method.
Alam
Syed Imteyaz
SI
Division of Biotechnology, Defence R & Establishment, Jhansi Road, Gwalior 474 002, India.
Agarwal
Gauri Shankar
GS
Kamboj
Dev Vrat
DV
Rai
Ganga Prasad
GP
Singh
Lokendra
L
D000881:Anthrax / Q000175:diagnosis / Q000382:microbiology; D001408:Bacillus anthracis / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis*; D004777:Environment; D004783:Environmental Microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D013171:Spores, Bacterial / Q000235:genetics / Q000302:isolation & purification*
D004269:DNA, Bacterial