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Volume 8, Number 10—October 2002
THEME ISSUE
Bioterrorism-related Anthrax
Bioterrorism-related Anthrax

Molecular Subtyping of Bacillus anthracis and the 2001 Bioterrorism-Associated Anthrax Outbreak, United States

Alex R. Hoffmaster*Comments to Author , Collette C. Fitzgerald*, Efrain Ribot*, Leonard W. Mayer*, and Tanja Popovic*
Author affiliations: *Centers for Disease Control and Prevention, Atlanta, Georgia, USA;

Main Article

Table 3

Molecular subtyping by MLVA and pagA sequencing performed on 28 clinical specimens from seven patients with inhalational anthraxa,b

Patient no.b Specimen typec Interval after antimicrobial therapy (days)d MLVA loci detected pagA Bacillus anthracis LRN PCRe
1 Pleural fluid 4 All negative Negative Positive
Pleural fluid 4 All negative Negative Positive
Blood 4 All negative Negative Negative
Lung 4 All negative ND ND
Lung 4 vrrA ND Negative
Heart blood 4 vrrA, vrrB1 Negative Negative
Pericardial blood 4 All negative Negative Positive
2 Thoracentesis fluid 4 vrrA Negative Positive
Serum 10 All negative Negative Positive
Respiratory wash 4 All negative Negative Positive
Pleural fluid 4 All negative Negative Positive
3 Blood culture 0 All negative Negative Positive
5 Blood culture 0 All negative Negative Positive
Blood culture 0 All negative Negative Positive
6 Blood culture 0 vrrA, vrrB1, vrrB2,,vrrC2 Negative Positive
10 Pleural fluid 1 vrrA, vrrB1, vrrB2, vrrC1, vrrC2, CG3, pXO1, pXO2 Positive Positive
Pleural fluid 1 vrrA, vrrB1, vrrB2, vrrC1, vrrC2, CG3, pXO1, pXO2 Positive Positive
Blood 1 All negative Negative Positive
CSF 3 All negative Negative Positive
Lung 3 vrrA, CG3 Negative Positive
Lymph node 3 vrrA, vrrB1, vrrB2, vrrC1, vrrC2, CG3, pXO1, pXO2 Positive Positive
11 Pleural fluid 2 All negative Positive Positive
Blood 2 All negative Negative Negative
Blood culture -1 All negative ND ND
Blood culture -1 All negative ND Positive
Blood culture -1 All negative ND ND
Blood culture -1 All negative ND Positive
Lymph node 4 vrrA, vrrB1, vrrB2, vrrC1, vrrC2, CG3 Positive Positive

aMLVA, multiple-locus variable-number tandem repeat analysis; pagA, protective antigen gene; LRN, Laboratory Response Network; PCR, polymerase chain reaction.
bPatients 1–10 described in Jernigan et al. (19) and patient 11 in Barakat et al (20)
cSpecimens collected postmortem.
dNumber of days the specimen was collected following or before the initiation of antimicrobial therapy. Specimens collected the same day as the initiation of therapy were designated as day 0 but were collected before antibiotic therapy.
eResults using the Laboratory Response Network PCR assay for detection of B. anthracis during the outbreak (18).

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References
  1. Harrell  LJ, Andersen  GL, Wilson  KH. Genetic variability of Bacillus anthracis and related species. J Clin Microbiol. 1995;33:184750.PubMedGoogle Scholar
  2. Henderson  ID, Yu  D, Turnbull  PC. Differentiation of Bacillus anthracis and other 'Bacillus cereus group' bacteria using IS231-derived sequences. FEMS Microbiol Lett. 1995;128:1138. DOIPubMedGoogle Scholar
  3. Helgason  E, Okstad  OA, Caugant  DA, Johansen  HA, Fouet  A, Mock  M, Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis—one species on the basis of genetic evidence. Appl Environ Microbiol. 2000;66:262730. DOIPubMedGoogle Scholar
  4. Keim  P, Kalif  A, Schupp  J, Hill  K, Travis  SE, Richmond  K, Molecular evolution and diversity in Bacillus anthracis as detected by amplified fragment length polymorphism markers. J Bacteriol. 1997;179:81824.PubMedGoogle Scholar
  5. Ticknor  LO, Kolsto  AB, Hill  KK, Keim  P, Laker  MT, Tonks  M, Fluorescent amplified fragment length polymorphism analysis of Norwegian Bacillus cereus and Bacillus thuringiensis soil isolates. Appl Environ Microbiol. 2001;67:486373. DOIPubMedGoogle Scholar
  6. Keim  P, Price  LB, Klevytska  AM, Smith  KL, Schupp  JM, Okinaka  R, Multiple-locus variable-number tandem repeat analysis reveals genetic relationships within Bacillus anthracis. J Bacteriol. 2000;182:292836. DOIPubMedGoogle Scholar
  7. Smith  KL, DeVos  V, Bryden  H, Price  LB, Hugh-Jones  ME, Keim  P. Bacillus anthracis diversity in Kruger National Park. J Clin Microbiol. 2000;38:37804.PubMedGoogle Scholar
  8. Price  LB, Hugh-Jones  M, Jackson  PJ, Keim  P. Genetic diversity in the protective antigen gene of Bacillus anthracis. J Bacteriol. 1999;181:235862.PubMedGoogle Scholar
  9. Khan  AS, Morse  S, Lillibridge  SR. Public-health preparedness for biological terrorism in the USA. Lancet. 2000;356:117982. DOIPubMedGoogle Scholar
  10. Logan  NA, Turnbull  PC. Bacillus and recently derived genera. In: Murray P, editor. Manual of clinical microbiology. Washington: ASM Press; 1991. p. 357–69.
  11. McBride  BW, Mogg  A, Telfer  JL, Lever  MS, Miller  J, Turnbull  PC, Protective efficacy of a recombinant protective antigen against Bacillus anthracis challenge and assessment of immunological markers. Vaccine. 1998;16:8107. DOIPubMedGoogle Scholar
  12. Barnard  JP, Friedlander  AM. Vaccination against anthrax with attenuated recombinant strains of Bacillus anthracis that produce protective antigen. Infect Immun. 1999;67:5627.PubMedGoogle Scholar
  13. Little  SF, Ivins  BE, Fellows  PF, Friedlander  AM. Passive protection by polyclonal antibodies against Bacillus anthracis infection in guinea pigs. Infect Immun. 1997;65:51715.PubMedGoogle Scholar
  14. Welkos  S, Little  S, Friedlander  A, Fritz  D, Fellows  P. The role of antibodies to Bacillus anthracis and anthrax toxin components in inhibiting the early stages of infection by anthrax spores. Microbiol. 2001;147:167785.
  15. Ivins  BE, Pitt  ML, Fellows  PF, Farchaus  JW, Benner  GE, Waag  DM, Comparative efficacy of experimental anthrax vaccine candidates against inhalation anthrax in rhesus macaques. Vaccine. 1998;16:11418. DOIPubMedGoogle Scholar
  16. Ivins  B, Fellows  P, Pitt  L, Estep  J, Farchaus  J, Friedlander  A, Experimental anthrax vaccines: efficacy of adjuvants combined with protective antigen against an aerosol Bacillus anthracis spore challenge in guinea pigs. Vaccine. 1995;13:177984. DOIPubMedGoogle Scholar
  17. Read  TD, Salzberg  SL, Pop  M, Shumway  M, Umayam  L, Jiang  L, Comparative Genome Sequenincg for Discovery of Novel Polymorphisms in Bacillus anthracis. Science. 2002;296:202833. DOIPubMedGoogle Scholar
  18. Hoffmaster  AR, Meyer  RF, Bowen  M, Marston  CK, Weyant  RS, Barnett  GA, Evaluation and validation of a real-time PCR assay for rapid identification of Bacillus anthracis. Emerg Infect Dis. 2002;8:117882.PubMedGoogle Scholar
  19. Jernigan  JA, Stephens  DS, Ashford  DA, Omenaca  C, Topiel  MS, Galbraith  M, Bioterrorism-related inhalational anthrax: the first 10 cases reported in the United States. Emerg Infect Dis. 2001;7:93344.PubMedGoogle Scholar
  20. Barakat  LA, Quentzel  HL, Jernigan  JA, Kirschke  DL, Griffith  K, Spear  SM, Fatal inhalational anthrax in a 94-Year-Old Connecticut woman. JAMA. 2002;287:8638. DOIPubMedGoogle Scholar

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