Improvement in Laboratory Diagnosis of Wound Botulism and Tetanus among Injecting Illicit-Drug Users by Use of Real-Time PCR Assays for Neurotoxin Gene Fragments
Open Access
- 1 September 2005
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 43 (9), 4342-4348
- https://doi.org/10.1128/jcm.43.9.4342-4348.2005
Abstract
An upsurge in wound infections due to Clostridium botulinum and Clostridium tetani among users of illegal injected drugs (IDUs) occurred in the United Kingdom during 2003 and 2004. A real-time PCR assay was developed to detect a fragment of the neurotoxin gene of C. tetani (TeNT) and was used in conjunction with previously described assays for C. botulinum neurotoxin types A, B, and E (BoNTA, -B, and -E). The assays were sensitive, specific, rapid to perform, and applicable to investigating infections among IDUs using DNA extracted directly from wound tissue, as well as bacteria growing among mixed microflora in enrichment cultures and in pure culture on solid media. A combination of bioassay and PCR test results confirmed the clinical diagnosis in 10 of 25 cases of suspected botulism and two of five suspected cases of tetanus among IDUs. The PCR assays were in almost complete agreement with the conventional bioassays when considering results from different samples collected from the same patient. The replacement of bioassays by real-time PCR for the isolation and identification of both C. botulinum and C. tetani demonstrates a sensitivity and specificity similar to those of conventional approaches. However, the real-time PCR assays substantially improves the diagnostic process in terms of the speed of results and by the replacement of experimental animals. Recommendations are given for an improved strategy for the laboratory investigation of suspected wound botulism and tetanus among IDUs.Keywords
This publication has 21 references indexed in Scilit:
- A case of infant botulism with a possible link to infant formula milk powder: evidence for the presence of more than one strain of Clostridium botulinum in clinical specimens and foodJournal of Medical Microbiology, 2005
- Development and Application of Real-Time PCR Assays to Detect Fragments of theClostridium botulinumTypes A, B, and E Neurotoxin Genes for Investigation of Human Foodborne and Infant BotulismFoodborne Pathogens & Disease, 2004
- Wound botulism in the UK and IrelandJournal of Medical Microbiology, 2004
- The genome sequence of Clostridium tetani , the causative agent of tetanus diseaseProceedings of the National Academy of Sciences, 2003
- Lethal outbreak of infection with Clostridium novyi type A and other spore-forming organisms in Scottish injecting drug usersJournal of Medical Microbiology, 2002
- Bacillus cereus cellulitis from contaminated heroinJournal of Medical Microbiology, 2002
- Wound Botulism in California, 1951–1998: Recent Epidemic in Heroin InjectorsClinical Infectious Diseases, 2000
- Phylogeny and taxonomy of the food-borne pathogen Clostridium botulinum and its neurotoxinsJournal of Applied Microbiology, 1998
- Wound Botulism in a Patient with a Tooth Abscess: Case Report and ReviewClinical Infectious Diseases, 1993
- A Simple Method for Storage of Bacteria at — 76°CJournal of Applied Bacteriology, 1978