1887

Abstract

A fluorescence-based multiplex PCR was automated for the simultaneous detection of , and in clinical samples from patients with suspected meningitis. Sensitivity of one to two genome copies per 100 μl sample and specificity of 100 % for each organism were shown. Automation of DNA extraction, liquid handling, PCR and analysis are achieved on a single platform, which enables a high throughput and rapid turnaround of clinical samples that, in turn, leads to faster diagnosis. This is ultimately beneficial to the treatment of the patient and for public health management.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.05416-0
2004-02-01
2019-11-15
Loading full text...

Full text loading...

/deliver/fulltext/jmm/53/2/JM530206.html?itemId=/content/journal/jmm/10.1099/jmm.0.05416-0&mimeType=html&fmt=ahah

References

  1. Borrow, R., Guiver, M., Sadler, F., Kaczmarski, E. B. & Fox, A. J. ( 1998;). False positive diagnosis of meningococcal infection by the IS1106 PCR ELISA. FEMS Microbiol Lett 162, 215–218.[CrossRef]
    [Google Scholar]
  2. Clarke, S. C. ( 2002;). Nucleotide sequence-based typing of bacteria and the impact of automation. Bioessays 24, 858–862.[CrossRef]
    [Google Scholar]
  3. Clarke, S. C., Reid, J., Thom, L., Denham, B. C. & Edwards, G. F. S. ( 2002;). Laboratory confirmation of meningococcal disease in Scotland, 1993–9. J Clin Pathol 55, 32–36.[CrossRef]
    [Google Scholar]
  4. Corless, C. E., Guiver, M., Borrow, R., Edwards-Jones, V., Fox, A. J. & Kaczmarski, E. B. ( 2001;). Simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in suspected cases of meningitis and septicemia using real-time PCR. J Clin Microbiol 39, 1553–1558.[CrossRef]
    [Google Scholar]
  5. Diggle, M. A. & Clarke, S. C. ( 2002;). Semi-automation of the polymerase chain reaction for laboratory confirmation of meningococcal disease. Br J Biomed Sci 59, 137–140.
    [Google Scholar]
  6. Diggle, M. A., Edwards, G. F. S. & Clarke, S. C. ( 2001;). Automation of fluorescence-based PCR for confirmation of meningococcal disease. J Clin Microbiol 39, 4518–4519.[CrossRef]
    [Google Scholar]
  7. Guiver, M., Borrow, R., Marsh, J., Gray, S. J., Kaczmarski, E. B., Howells, D., Boseley, P. & Fox, A. J. ( 2000;). Evaluation of the Applied Biosystems automated Taqman polymerase chain reaction system for the detection of meningococcal DNA. FEMS Immunol Med Microbiol 28, 173–179.[CrossRef]
    [Google Scholar]
  8. Koedel, U., Scheld, W. M. & Pfister, H.-W. ( 2002;). Pathogenesis and pathophysiology of pneumococcal meningitis. Lancet Infect Dis 2, 721–736.[CrossRef]
    [Google Scholar]
  9. Peltola, H. ( 2000;). Worldwide Haemophilus influenzae type b disease at the beginning of the 21st century: global analysis of the disease burden 25 years after the use of the polysaccharide vaccine and a decade after the advent of conjugates. Clin Microbiol Rev 13, 302–317.[CrossRef]
    [Google Scholar]
  10. Rosenstein, N. E., Perkins, B. A., Stephens, D. S., Popovic, T. & Hughes, J. M. ( 2001;). Meningococcal disease. N Engl J Med 344, 1378–1388.[CrossRef]
    [Google Scholar]
  11. Smith, K., Diggle, M. A. & Clarke, S. C. ( 2003;). Comparison of commercial DNA extraction kits for extraction of bacterial genomic DNA from whole-blood samples. J Clin Microbiol 41, 2440–2443.[CrossRef]
    [Google Scholar]
  12. van Deuren, M., Brandtzaeg, P. & van der Meer, J. W. M. ( 2000;). Update on meningococcal disease with emphasis on pathogenesis and clinical management. Clin Microbiol Rev 13, 144–166.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.05416-0
Loading
/content/journal/jmm/10.1099/jmm.0.05416-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error