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Volume 18, Number 8—August 2012

Molecular Epidemiologic Investigation of an Anthrax Outbreak among Heroin Users, Europe

Erin P. Price1, Meagan L. Seymour, Derek S. Sarovich1, Jennie Latham, Spenser R. Wolken, Joanne Mason, Gemma Vincent, Kevin P. Drees, Stephen M. Beckstrom-Sternberg, Adam M. Phillippy, Sergey Koren, Richard T. Okinaka, Wai-Kwan Chung, James M. Schupp, David M. Wagner, Richard Vipond, Jeffrey T. Foster, Nicholas H. Bergman, James Burans, Talima Pearson, Tim Brooks, and Paul KeimComments to Author 
Author affiliations: Northern Arizona University, Flagstaff, Arizona, USA (E.P. Price, M.L. Seymour, D.S. Sarovich, S.R. Wolken, K.P. Drees, S.M. Beckstrom-Sternberg, R.T. Okinaka, W.-K. Chung, J.M. Schupp, D.M. Wagner, J.T. Foster, T. Pearson, P. Keim); Health Protection Agency, London, UK (J. Latham, J. Mason, G. Vincent, R. Vipond, T. Brooks); National Biodefense Analysis and Countermeasures Center, Frederick, Maryland, USA (A.M. Phillippy, S. Koren, N. H. Bergman, J. Burans); and Translational Genomics Research Institute, Phoenix, Arizona, USA (S.M. Beckstrom-Sternberg, J.M. Schupp, P. Keim)

Main Article


Genotyping results of 36 culture- or PCR-confirmed cases of anthrax in heroin users, 2009–2010, Europe*

Sample no. Status† Collection date Location SNP1053700 SNP1173928
Ames‡ NA 1981 Texas, USA A G
Ba4599§ C+/PCR+ 2009 Dec 16 Glasgow, Scotland G C
A112 C+/PCR+ 2009 Dec 18 Germany G C
4622 C+/PCR+ 2009 Dec 19 Glasgow, Scotland G C
4646 C-/PCR+ 2009 Dec 22 Airdrie, Scotland G C
4670 C+/PCR+ 2009 Dec 23 Glasgow, Scotland G C
4745 C-/PCR+ 2009 Dec 31 Glasgow, Scotland G C
0002 C+/PCR+ 2010 Jan 4 Glasgow, Scotland G C
0007 C+/PCR+ 2010 Jan 4 Glasgow, Scotland G C
0001 C+/PCR+ 2010 Jan 4 Dundee, Scotland G C
0046 C-/PCR+ 2010 Jan 6 Stirling, Scotland G C
0074 C+/PCR+ 2010 Jan 7 Kirkcaldy, Scotland G C
0075 C+/PCR+ 2010 Jan 7 Kirkcaldy, Scotland G C
0117(2) C+/PCR+ 2010 Jan 9 Dundee, Scotland G C
0142 C+/PCR+ 2010 Jan 9 Glasgow, Scotland G C
0271 C-/PCR+ 2010 Jan 15 Glasgow, Scotland G C
0393 C+/PCR+ 2010 Jan 20 Kilmarnock, Scotland G C
0426 C+/PCR+ 2010 Jan 21 Glasgow, Scotland G C
0491 C+/PCR+ 2010 Jan 22 Glasgow, Scotland G C
0773 C+/PCR+ 2010 Feb 4 London, England G C
0871 C+/PCR+ 2010 Feb 9 Glasgow, Scotland G C
0874 C+/PCR+ 2010 Feb 9 Glasgow, Scotland G C
0844 C-/PCR+ 2010 Feb 9 Blackpool, England G C
1297 C+/PCR+ 2010 Feb 26 London, England G C
1060 C+/PCR+ 2010 Mar 5 Fife, Scotland G C
1134 C+/PCR+ 2010 Mar 5 Dumfries, Scotland G C
1320 C+/PCR+ 2010 Mar 5 Dumfries, Scotland G C
0981 C-/PCR+ 2010 Mar 5 Airdrie, Scotland G C
1458 C-/PCR+ 2010 Apr 1 Dumfries, Scotland G C
1927 C-/PCR+ 2010 Apr 1 Paisley, Scotland G C
2145 C-/PCR+ 2010 Apr 16 Livingston, Scotland G C
2199 C-/PCR+ 2010 Apr 16 Glasgow, Scotland G C
2506 C+/PCR+ 2010 Apr 27 Edinburgh, Scotland G C
2728 C+/PCR+ 2010 May 7 Glasgow, Scotland G C
3739 C+/PCR+ 2010 Jul 16 Glasgow, Scotland G C
4936/5011 C-/PCR+ 2010 Aug 26 Leicester, England G C
6696 C+/PCR+ 2010 Nov 1 Maidstone, England G C

*ID, identification; SNP, single nucleotide polymorphism; NA, not applicable; C, culture. Boldface indicates pairs of isolates from the same patient.
†PCR+ samples were determined by using assays targeting cap, lef (Special Pathogens Reference Unit (UK), pers. comm.) and bagC loci (14).
‡Ancestral allele DNA control.
§Derived allele DNA control.

Main Article

  1. Van Ert  MN, Easterday  WR, Huynh  LY, Okinaka  RT, Hugh-Jones  ME, Ravel  J, Global genetic population structure of Bacillus anthracis. PLoS ONE. 2007;2:e461. DOIPubMedGoogle Scholar
  2. Keim  P, Smith  KL. Bacillus anthracis evolution and epidemiology. Curr Top Microbiol Immunol. 2002;271:2132.PubMedGoogle Scholar
  3. Pearson  T, Busch  JD, Ravel  J, Read  TD, Rhoton  SD, U’Ren  JM, Phylogenetic discovery bias in Bacillus anthracis using single-nucleotide polymorphisms from whole-genome sequencing. Proc Natl Acad Sci U S A. 2004;101:1353641. DOIPubMedGoogle Scholar
  4. Kenefic  LJ, Pearson  T, Okinaka  RT, Schupp  JM, Wagner  DM, Hoffmaster  AR, Pre-Columbian origins for North American anthrax. PLoS ONE. 2009;4:e4813. DOIPubMedGoogle Scholar
  5. Mayer  TA, Bersoff-Matcha  S, Murphy  C, Earls  J, Harper  S, Pauze  D, Clinical presentation of inhalational anthrax following bioterrorism exposure: report of 2 surviving patients. JAMA. 2001;286:254953. DOIPubMedGoogle Scholar
  6. Ringertz  SH, Hoiby  EA, Jensenius  M, Maehlen  J, Caugant  DA, Myklebust  A, Injectional anthrax in a heroin skin-popper. Lancet. 2000;356:15745. DOIPubMedGoogle Scholar
  7. Knox  D, Murray  G, Millar  M, Hamilton  D, Connor  M, Ferdinand  RD, Subcutaneous anthrax in three intravenous drug users: a new clinical diagnosis. J Bone Joint Surg Br. 2011;93-B:4147. DOIPubMedGoogle Scholar
  8. Ramsay  CN, Stirling  A, Smith  J, Hawkins  G, Brooks  T, Hood  J, An outbreak of infection with Bacillus anthracis in injecting drug users in Scotland. Euro Surveill. 2010;15:pii:19465.PubMedGoogle Scholar
  9. National Health Service. An outbreak of anthrax among drug users in Scotland, December 2009 to December 2010: a report on behalf of the National Anthrax Outbreak Control Team. December 2011 [cited 2012 Jan 24].
  10. European Centre for Disease Prevention and Control. Annual epidemiological report on communicable diseases in Europe 2010 [cited 2011 Jul 19].
  11. Powell  AG, Crozier  JE, Hodgson  H, Galloway  DJ. A case of septicaemic anthrax in an intravenous drug user. BMC Infect Dis. 2011;11:21. DOIPubMedGoogle Scholar
  12. Booth  MG, Hood  J, Brooks  TJ, Hart  A. Anthrax infection in drug users. Lancet. 2010;375:13456. DOIPubMedGoogle Scholar
  13. Keim  P, Van Ert  MN, Pearson  T, Vogler  AJ, Huynh  LY, Wagner  DM. Anthrax molecular epidemiology and forensics: using the appropriate marker for different evolutionary scales. Infect Genet Evol. 2004;4:20513. DOIPubMedGoogle Scholar
  14. Radun  D, Bernard  H, Altmann  M, Schoneberg  I, Bochat  V, van Treeck  U, Preliminary case report of fatal anthrax in an injecting drug user in North-Rhine-Westphalia, Germany, December 2009. Euro Surveill. 2010;15:pii: 19464.PubMedGoogle Scholar
  15. Okinaka  RT, Henrie  M, Hill  KK, Lowery  KS, Van Ert  M, Pearson  T, Single nucleotide polymorphism typing of Bacillus anthracis from Sverdlovsk tissue. Emerg Infect Dis. 2008;14:6536. DOIPubMedGoogle Scholar
  16. de Lamballerie  X, Zandotti  C, Vignoli  C, Bollet  C, de Micco  P. A one-step microbial DNA extraction method using “Chelex 100” suitable for gene amplification. Res Microbiol. 1992;143:78590. DOIPubMedGoogle Scholar
  17. Marston  CK, Allen  CA, Beaudry  J, Price  EP, Wolken  SR, Pearson  T, Molecular epidemiology of anthrax cases associated with recreational use of animal hides and yarn in the United States. PLoS ONE. 2011;6:e28274. DOIPubMedGoogle Scholar
  18. Newton  CR, Graham  A, Heptinstall  LE, Powell  SJ, Summers  C, Kalsheker  N, Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Res. 1989;17:250316. DOIPubMedGoogle Scholar
  19. Rhodes  RB, Lewis  K, Shultz  J, Huber  S, Voelkerding  KV, Leonard  DG, Analysis of the factor V Leiden mutation using the READIT Assay. Mol Diagn. 2001;6:5561. DOIPubMedGoogle Scholar
  20. Van Ert  MN, Easterday  WR, Simonson  TS, U’Ren  JM, Pearson  T, Kenefic  LJ, Strain-specific single-nucleotide polymorphism assays for the Bacillus anthracis Ames strain. J Clin Microbiol. 2007;45:4753. DOIPubMedGoogle Scholar
  21. Miller  JR, Delcher  AL, Koren  S, Venter  E, Walenz  BP, Brownley  A, Aggressive assembly of pyrosequencing reads with mates. Bioinformatics. 2008;24:281824. DOIPubMedGoogle Scholar
  22. Li  H, Durbin  R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009;25:175460. DOIPubMedGoogle Scholar
  23. Li  H, Durbin  R. Fast and accurate long-read alignment with Burrows-Wheeler transform. Bioinformatics. 2010;26:58995. DOIPubMedGoogle Scholar
  24. Li  H, Handsaker  B, Wysoker  A, Fennell  T, Ruan  J, Homer  N, The Sequence Alignment/Map format and SAMtools. Bioinformatics. 2009;25:20789. DOIPubMedGoogle Scholar
  25. Kurtz  S, Phillippy  A, Delcher  AL, Smoot  M, Shumway  M, Antonescu  C, Versatile and open software for comparing large genomes. Genome Biol. 2004;5:R12. DOIPubMedGoogle Scholar
  26. Swofford  DL. PAUP*: phylogenetic analysis using parsimony (*and other methods), version 4.0b10. Sunderland (MA): Sinauer Associates; 2002.
  27. United Nations Office on Drugs and Crime. Afghanistan opium survey 2009: summary findings [cited 2011 Jul 20].
  28. Chouvy  P-A. Opium: uncovering the politics of the poppy. Cambridge (MA): Harvard University Press; 2010.
  29. McNeil  DG Jr. Anthrax: in Scotland, six heroin users die of anthrax poisoning. New York Times. 201l Jan 12 [cited 2011 Jul 20].
  30. Manchee  RJ, Broster  MG, Melling  J, Henstridge  RM, Stagg  AJ. Bacillus anthracis on Gruinard Island. Nature. 1981;294:2545. DOIPubMedGoogle Scholar
  31. Interpol. Heroin. 2010 May 18 [cited 2011 Jul 20].
  32. Boekhout van Solinge  T. Drug use and drug trafficking in Europe. Tijdschr Econ Soc Geogr. 1998;89:1005. DOIGoogle Scholar
  33. Centers for Disease Control and Prevention. Gastrointestinal anthrax after an animal-hide drumming event—New Hampshire and Massachusetts, 2009. MMWR Morb Mortal Wkly Rep. 2010;59:8727.PubMedGoogle Scholar
  34. Centers for Disease Control and Prevention. Cutaneous anthrax associated with drum making using goat hides from West Africa–Connecticut, 2007. MMWR Morb Mortal Wkly Rep. 2008;57:62831.PubMedGoogle Scholar
  35. Centers for Disease Control and Prevention. Inhalation anthrax associated with dried animal hides–Pennsylvania and New York City, 2006. MMWR Morb Mortal Wkly Rep. 2006;55:2802.PubMedGoogle Scholar
  36. Centers for Disease Control and Prevention. Human cutaneous anthrax—North Carolina, 1987. MMWR Morb Mortal Wkly Rep. 1988;37:4134.PubMedGoogle Scholar
  37. Anaraki  S, Addiman  S, Nixon  G, Krahe  D, Ghosh  R, Brooks  T, Investigations and control measures following a case of inhalation anthrax in East London in a drum maker and drummer, October 2008. Euro Surveill. 2008;13:pii:19076.PubMedGoogle Scholar
  38. Eurosurveillance Editorial Team. Probable human anthrax death in Scotland. Euro Surveill. 2006;11:E060817.2.
  39. Antwerpen  MH, Zimmermann  P, Bewley  K, Frangoulidis  D, Meyer  H. Real-time PCR system targeting a chromosomal marker specific for Bacillus anthracis. Mol Cell Probes. 2008;22:3135. DOIPubMedGoogle Scholar

Main Article

1Current affiliation: Menzies School of Health Research, Casuarina, Northern Territory, Australia.

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