Academic Journal
Automated Detection of Toxigenic Clostridium difficile in Clinical Samples: Isothermal tcdB Amplification Coupled to Array-Based Detection
| العنوان: | Automated Detection of Toxigenic Clostridium difficile in Clinical Samples: Isothermal tcdB Amplification Coupled to Array-Based Detection |
|---|---|
| المؤلفون: | Hicke, Brian, Pasko, Chris, Groves, Benjamin, Ager, Edward, Corpuz, Maylene, Frech, Georges, Munns, Denton, Smith, Wendy, Warcup, Ashley, Denys, Gerald, Ledeboer, Nathan A., Lindsey, Wes, Owen, Charles, Rea, Larry, Jenison, Robert |
| المصدر: | Journal of Clinical Microbiology ; volume 50, issue 8, page 2681-2687 ; ISSN 0095-1137 1098-660X |
| بيانات النشر: | American Society for Microbiology |
| سنة النشر: | 2012 |
| الوصف: | Clostridium difficile can carry a genetically variable pathogenicity locus (PaLoc), which encodes clostridial toxins A and B. In hospitals and in the community at large, this organism is increasingly identified as a pathogen. To develop a diagnostic test that combines the strengths of immunoassays (cost) and DNA amplification assays (sensitivity/specificity), we targeted a genetically stable PaLoc region, amplifying tcdB sequences and detecting them by hybridization capture. The assay employs a hot-start isothermal method coupled to a multiplexed chip-based readout, creating a manual assay that detects toxigenic C. difficile with high sensitivity and specificity within 1 h. Assay automation on an electromechanical instrument produced an analytical sensitivity of 10 CFU (95% probability of detection) of C. difficile in fecal samples, along with discrimination against other enteric bacteria. To verify automated assay function, 130 patient samples were tested: 31/32 positive samples (97% sensitive; 95% confidence interval [CI], 82 to 99%) and 98/98 negative samples (100% specific; 95% CI, 95 to 100%) were scored correctly. Large-scale clinical studies are now planned to determine clinical sensitivity and specificity. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1128/jcm.00621-12 |
| DOI: | 10.1128/JCM.00621-12 |
| الاتاحة: | http://dx.doi.org/10.1128/jcm.00621-12 https://journals.asm.org/doi/pdf/10.1128/JCM.00621-12 |
| Rights: | https://journals.asm.org/non-commercial-tdm-license |
| رقم الانضمام: | edsbas.1DBE8D69 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1128/jcm.00621-12 |
|---|