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Journal of Clinical Microbiology, July 2004, p. 2980-2987, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.2980-2987.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Use of Robotized DNA Isolation and Real-Time PCR To Quantify and Identify Close Correlation between Levels of Neisseria meningitidis DNA and Lipopolysaccharides in Plasma and Cerebrospinal Fluid from Patients with Systemic Meningococcal Disease

Reidun Øvstebø,1* Petter Brandtzaeg,2 Berit Brusletto,1 Kari Bente Foss Haug,1 Knut Lande,1 Ernst Arne Høiby,3 and Peter Kierulf1

Departments of Clinical Chemistry,1 Pediatrics, Ullevål University Hospital,2 Norwegian Institute of Public Health, Oslo, Norway3

Received 13 January 2004/ Returned for modification 3 March 2004/ Accepted 4 April 2004

The present study, using robotized DNA isolation and quantitative PCR based on the Neisseria meningitidis-specific capsular transport A gene, demonstrates the ease, rapidity, specificity, and sensitivity of quantifying neisserial DNA in plasma (n = 65) and cerebrospinal fluid (CSF) (n = 12) from patients with systemic meningococcal disease. We found a close correlation between the levels of neisserial DNA and lipopolysaccharides in plasma (r = 0.905) and in CSF (r = 0.964). The median concentration of neisserial DNA in plasma in 23 patients with persistent shock was 2 x 107 copies/ml, versus <103 copies/ml in 42 nonshock patients. Furthermore, quantitative PCR made possible estimates of the total number of meningococci in plasma, as opposed to conventional blood cultures, suggesting about 1,000 dead meningococci for every viable bacterium. Finally, with logistic regression analyses, neisserial DNA may predict a patient's disease severity and outcome at hospital admission. The number of meningococci in plasma and CSF appears to be the main determinant of the lipopolysaccharide levels, clinical presentation, and outcome.

* Corresponding author. Mailing address: The Research and Development Group, Department of Clinical Chemistry, Ullevål University Hospital, NO-0407 Oslo, Norway. Phone: 47-22119490. Fax: 47-22118189. E-mail: reidun.ovstebo{at}medisin.uio.no.

Journal of Clinical Microbiology, July 2004, p. 2980-2987, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.2980-2987.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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