Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2004 Jul;42(7):2980-7.
doi: 10.1128/JCM.42.7.2980-2987.2004.

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, Berit Brusletto, Kari Bente Foss Haug, Knut Lande, Ernst Arne Høiby, Peter Kierulf
Affiliations

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 et al. J Clin Microbiol. 2004 Jul.

Erratum in

  • J Clin Microbiol. 2005 Jan;43(1):532

Abstract

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 10(7) copies/ml, versus <10(3) 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.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
Levels of neisserial DNA (a) and LPS (b) in plasma samples from patients with systemic meningococcal disease presenting with various clinical syndromes. The dotted lines indicate the detection limits, 103 neisserial DNA copies/ml of plasma and 0.25 IU of LPS/ml of plasma. Horizontal bars denote median values.
FIG. 2.
FIG. 2.
Levels of neisserial DNA (a) and LPS (b) in CSF samples from patients with systemic meningococcal disease presenting with various clinical syndromes. The dotted lines indicate the detection limits, 103 neisserial DNA copies/ml of CSF and 0.25 IU of LPS/ml of CSF.
FIG. 3.
FIG. 3.
Clearance of neisserial DNA (a) and LPS (b) in plasma samples collected after admission to the hospital from eight patients with fulminant meningococcal septicemia. The first sample (hour 0) was collected immediately before initiation of antibiotic therapy from seven of eight patients. The first sample from patient 7 was collected 1 h after the initiation of antibiotic therapy. Patients 2, 3, and 7 died. The half-life of neisserial DNA (a) was 3 to 4 h and that of LPS (b) was 1 to 3 h. The dotted lines indicate the detection limits, 103 neisserial DNA copies/ml of plasma and 0.25 IU of LPS/ml of plasma.
FIG. 4.
FIG. 4.
Relationship between neisserial DNA and LPS in plasma samples from patients with systemic meningococcal disease at admission to hospital. The dotted lines separate samples as either positive or negative for neisserial DNA or LPS. ▪, fulminant septicemia, fatal cases (n = 9); ▪, fulminant septicemia, survivors (n = 12); ×, septicemia and meningitis (n = 2); ○, mild systemic meningococcal disease (n = 14); ▵, distinct meningitis (n = 28).
See this image and copyright information in PMC

References

    1. Andersen, B. M., O. Solberg, K. Bryn, L. O. Froholm, P. Gaustad, E. A. Hoiby, B. E. Kristiansen, and K. Bovre. 1987. Endotoxin liberation from Neisseria meningitidis isolated from carriers and clinical cases. Scand. J. Infect. Dis. 19:409-419. - PubMed
    1. Bjerre, A., B. Brusletto, T. E. Mollnes, E. Fritzsøonn, E. Rosenqvist, E. Wedege, E. Namork, P. Kierulf, and P. Brandtzaeg. 2002. Complement activation induced by purified Neisseria meningitidis lipopolysaccharides (LPS), outer membrane vesicles, whole bacteria, and an LPS-free mutant. J. Infect. Dis. 185:220-228. - PubMed
    1. Bjerre, A., B. Brusletto, R. Ovstebo, G. B. Joo, P. Kierulf, and P. Brandtzaeg. 2003. Identification of meningococcal LPS as a major monocyte activator in IL-10 depleted shock plasmas and CSF by blocking the CD14-TLR4 receptor complex. J. Endotoxin Res. 9:155-163. - PubMed
    1. Bjerre, A., B. Brusletto, E. Rosenqvist, E. Namork, P. Kierulf, R. Ovstebo, G. B. Joo, and P. Brandtzæg. 2000. Cellular activation properties and morphology of membrane-bound and purified meningococcal lipopolysaccharide. J. Endotoxin Res. 6:437-445. - PubMed
    1. Brandtzaeg, P. 2003. Host response to Neisseria meningitidis lacking lipopolysaccharides. Expert Rev. Anti-infect. Ther. 1:89-96. - PubMed

MeSH terms