Mass spectrometry provides accurate characterization of two genetic marker types in Bacillus anthracis.
Epidemiological and forensic analyses of bioterrorism events involving Bacillus anthracis could be improved if both variable number tandem repeats (VNTRs) and single nucleotide polymorphisms (SNPs) could be combined on a single analysis platform. Here we present the use of electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS) to characterize 24 alleles from 6 VNTR loci and 11 alleles from 7 SNP loci in B. anthracis. The results obtained with ESI-FTICR-MS were consistent with independent results obtained from traditional approaches using electrophoretic detection of fluorescent products. However, ESI-FTICR-MS improves on the traditional approaches because it does not require fluorescent labeling of PCR products, minimizes post-PCR processing, obviates electrophoresis, and provides unambiguous base composition of both SNP and VNTR PCR products. In addition, ESI-FTICR-MS allows both marker types to be examined simultaneously and at a rate of approximately 1 sample per min. This technology represents a significant advance in our ability to rapidly characterize B. anthracis isolates using VNTR and SNP loci.
Van Ert
Matthew N
MN
Northern Arizona University, Flagstaff, AZ 86011-5640, USA.
Hofstadler
Steven A
SA
Jiang
Yun
Y
Busch
Joseph D
JD
Wagner
David M
DM
Drader
Jared J
JJ
Ecker
David J
DJ
Hannis
James C
JC
Huynh
Lynn Y
LY
Schupp
James M
JM
Simonson
Tatum S
TS
Keim
Paul
P
D000483:Alleles; D001408:Bacillus anthracis / Q000737:chemistry / Q000235:genetics*; D023482:Bioterrorism; D005583:Fourier Analysis; D005819:Genetic Markers; D005838:Genotype; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D020080:Tandem Repeat Sequences
D005819:Genetic Markers