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Published in: Intensive Care Medicine 7/2007

01-07-2007 | Pediatric Original

Detection of Neisseria meningitidis DNA from skin lesion biopsy using real-time PCR: usefulness in the aetiological diagnosis of purpura fulminans

Authors: Pierre Staquet, Ludovic Lemee, Eva Verdier, Guy Bonmarchand, Vincent Laudenbach, Christine Michel, Jean-Francois Lemeland, Stephane Marret, Thierry Blanc

Published in: Intensive Care Medicine | Issue 7/2007

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Abstract

Objective

The present study evaluated the usefulness of a real-time polymerase chain reaction (rtPCR) assay for the detection of Neisseria meningitidis (Nm) and genogrouping on skin lesion biopsies in patients with purpura fulminans (PF).

Design

Retrospective single-centre study.

Setting

Adult and paediatric intensive care units at the University Hospital of Rouen.

Patients

All patients admitted between January 2000 and January 2006, with a final diagnosis of PF and for which a skin biopsy and blood cultures were performed, were included.

Interventions

Skin biopsy and blood cultures were used for culture and rtPCR.

Measurements and main results

Thirty-four patients fulfilled the criteria (27 children and 7 adults). Nm rtPCR performed on skin biopsy was positive in 100% (34/34) of cases, compared with only 14.7% (5/34) for skin culture (p = 0.0001). rtPCR genogrouping on skin biopsy was positive in 58.8% (20/34) of the cases compared with 14.7% (5/34) for skin culture (p = 0.0013). For patients (n = 17) in whom rtPCR was performed both on blood and skin biopsy, skin biopsy gave a significantly higher rate of Nm detection [100% (17/17) vs. 58.8% (10/17); p = 0.023] and genogroup characterisation [76.5% (13/17) vs. 35.3% (6/17); p = 0.045] than blood. We encountered no specimen with culture-positive and rtPCR-negative results (negative predictive value of rtPCR 100%).

Conclusion

In suspected PF cases, skin biopsy is more reliable to identify Nm and its genogroup than blood or, probably, CSF, especially when PCR methods are used. This could help the implementation of public health interventions, especially concerning a vaccination policy.
Literature
1.
go back to reference Corless CE, Guiver M, Borrow R, Edwards-Jones V, Fox AJ, Kaczmarski EB (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 39:1553–1558CrossRefPubMedPubMedCentral Corless CE, Guiver M, Borrow R, Edwards-Jones V, Fox AJ, Kaczmarski EB (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 39:1553–1558CrossRefPubMedPubMedCentral
2.
go back to reference Kaczmarski EB (1997) Meningococcal disease in England and Wales: 1995. Commun Dis Rep CDR Rev 7:R55–59PubMed Kaczmarski EB (1997) Meningococcal disease in England and Wales: 1995. Commun Dis Rep CDR Rev 7:R55–59PubMed
3.
go back to reference Kaczmarski EB, Ragunathan PL, Marsh J, Gray SJ, Guiver M (1998) Creating a national service for the diagnosis of meningococcal disease by polymerase chain reaction. Commun Dis Public Health 1:54–56PubMed Kaczmarski EB, Ragunathan PL, Marsh J, Gray SJ, Guiver M (1998) Creating a national service for the diagnosis of meningococcal disease by polymerase chain reaction. Commun Dis Public Health 1:54–56PubMed
4.
go back to reference Perrocheau A (2004) Les infections invasives à méningocoques en France, en 2003. Bulletin Epidémiologique Hebdomadaire 46:217–220 Perrocheau A (2004) Les infections invasives à méningocoques en France, en 2003. Bulletin Epidémiologique Hebdomadaire 46:217–220
5.
go back to reference Guiver M, Borrow R, Marsh J, Gray SJ, Kaczmarski EB, Howells D, Boseley P, Fox AJ (2000) Evaluation of the Applied Biosystems automated Taqman polymerase chain reaction system for the detection of meningococcal DNA. FEMS Immunol Med Microbiol 28:173–179CrossRefPubMed Guiver M, Borrow R, Marsh J, Gray SJ, Kaczmarski EB, Howells D, Boseley P, Fox AJ (2000) Evaluation of the Applied Biosystems automated Taqman polymerase chain reaction system for the detection of meningococcal DNA. FEMS Immunol Med Microbiol 28:173–179CrossRefPubMed
6.
go back to reference Taha MK (2000) Simultaneous approach for nonculture PCR-based identification and serogroup prediction of Neisseria meningitidis. J Clin Microbiol 38:855–857PubMedPubMedCentral Taha MK (2000) Simultaneous approach for nonculture PCR-based identification and serogroup prediction of Neisseria meningitidis. J Clin Microbiol 38:855–857PubMedPubMedCentral
7.
go back to reference van Deuren M, van Dijke BJ, Koopman RJ, Horrevorts AM, Meis JF, Santman FW, van der Meer JW (1993) Rapid diagnosis of acute meningococcal infections by needle aspiration or biopsy of skin lesions. BMJ 306:1229–1232CrossRefPubMedPubMedCentral van Deuren M, van Dijke BJ, Koopman RJ, Horrevorts AM, Meis JF, Santman FW, van der Meer JW (1993) Rapid diagnosis of acute meningococcal infections by needle aspiration or biopsy of skin lesions. BMJ 306:1229–1232CrossRefPubMedPubMedCentral
8.
go back to reference Periappuram M, Taylor MR, Keane CT (1995) Rapid detection of meningococci from petechiae in acute meningococcal infection. J Infect 31:201–203CrossRefPubMed Periappuram M, Taylor MR, Keane CT (1995) Rapid detection of meningococci from petechiae in acute meningococcal infection. J Infect 31:201–203CrossRefPubMed
9.
go back to reference Staquet P, Lemee L, Verdier E, Chadie A, Michel C, Laudenbach V, Marret S, Blanc T (2006) Detection of Neisseria meningitidis DNA from skin lesion biopsy using real-time PCR: Usefulness in the etiological diagnosis of purpura fulminans. European Academy Of Paediatrics, Barcelona, Spain, October 7–10, 2006 Staquet P, Lemee L, Verdier E, Chadie A, Michel C, Laudenbach V, Marret S, Blanc T (2006) Detection of Neisseria meningitidis DNA from skin lesion biopsy using real-time PCR: Usefulness in the etiological diagnosis of purpura fulminans. European Academy Of Paediatrics, Barcelona, Spain, October 7–10, 2006
10.
go back to reference Leclerc F, Noizet O, Dorkenoo A, Cremer R, Leteurtre S, Sadik A, Fourier C (2001) [Treatment of meningococcal purpura fulminans]. Arch Pediatr 8 Suppl 4:677s–688sCrossRef Leclerc F, Noizet O, Dorkenoo A, Cremer R, Leteurtre S, Sadik A, Fourier C (2001) [Treatment of meningococcal purpura fulminans]. Arch Pediatr 8 Suppl 4:677s–688sCrossRef
11.
go back to reference Public Health Laboratory Service, P. H. M. E. G, Scottish Centre for Infection and Environmental Health (2002) Guidelines for public health management of meningococcal disease in the UK. Commun Dis Public Health 5:187–204 Public Health Laboratory Service, P. H. M. E. G, Scottish Centre for Infection and Environmental Health (2002) Guidelines for public health management of meningococcal disease in the UK. Commun Dis Public Health 5:187–204
12.
go back to reference Olsen KM, Peddicord TE, Campbell GD, Rupp ME (2000) Antimicrobial effects of lidocaine in bronchoalveolar lavage fluid. J Antimicrob Chemother 45:217–219CrossRefPubMed Olsen KM, Peddicord TE, Campbell GD, Rupp ME (2000) Antimicrobial effects of lidocaine in bronchoalveolar lavage fluid. J Antimicrob Chemother 45:217–219CrossRefPubMed
13.
go back to reference Aydin ON, Eyigor M, Aydin N (2001) Antimicrobial activity of ropivacaine and other local anaesthetics. Eur J Anaesthesiol 18:687–694CrossRefPubMed Aydin ON, Eyigor M, Aydin N (2001) Antimicrobial activity of ropivacaine and other local anaesthetics. Eur J Anaesthesiol 18:687–694CrossRefPubMed
14.
go back to reference Ploy MC, Garnier F, Languepin J, Fermeaux V, Martin C, Denis F (2005) Interest of postmortem-collected specimens in the diagnosis of fulminant meningococcal sepsis. Diagn Microbiol Infect Dis 52:65–66CrossRefPubMed Ploy MC, Garnier F, Languepin J, Fermeaux V, Martin C, Denis F (2005) Interest of postmortem-collected specimens in the diagnosis of fulminant meningococcal sepsis. Diagn Microbiol Infect Dis 52:65–66CrossRefPubMed
15.
go back to reference Guarner J, Greer PW, Whitney A, Shieh WJ, Fischer M, White EH, Carlone GM, Stephens DS, Popovic T, Zaki SR (2004) Pathogenesis and diagnosis of human meningococcal disease using immunohistochemical and PCR assays. Am J Clin Pathol 122:754–764CrossRefPubMed Guarner J, Greer PW, Whitney A, Shieh WJ, Fischer M, White EH, Carlone GM, Stephens DS, Popovic T, Zaki SR (2004) Pathogenesis and diagnosis of human meningococcal disease using immunohistochemical and PCR assays. Am J Clin Pathol 122:754–764CrossRefPubMed
16.
go back to reference Taha MK, Alonso JM, Cafferkey M, Caugant DA, Clarke SC, Diggle MA, Fox A, Frosch M, Gray SJ, Guiver M, Heuberger S, Kalmusova J, Kesanopoulos K, Klem AM, Kriz P, Marsh J, Molling P, Murphy K, Olcen P, Sanou O, Tzanakaki G, Vogel U (2005) Interlaboratory comparison of PCR-based identification and genogrouping of Neisseria meningitidis. J Clin Microbiol 43:144–149CrossRefPubMedPubMedCentral Taha MK, Alonso JM, Cafferkey M, Caugant DA, Clarke SC, Diggle MA, Fox A, Frosch M, Gray SJ, Guiver M, Heuberger S, Kalmusova J, Kesanopoulos K, Klem AM, Kriz P, Marsh J, Molling P, Murphy K, Olcen P, Sanou O, Tzanakaki G, Vogel U (2005) Interlaboratory comparison of PCR-based identification and genogrouping of Neisseria meningitidis. J Clin Microbiol 43:144–149CrossRefPubMedPubMedCentral
17.
go back to reference Birtles A, Hardy K, Gray SJ, Handford S, Kaczmarski EB, Edwards-Jones V, Fox AJ (2005) Multilocus sequence typing of Neisseria meningitidis directly from clinical samples and application of the method to the investigation of meningococcal disease case clusters. J Clin Microbiol 43:6007–6014CrossRefPubMedPubMedCentral Birtles A, Hardy K, Gray SJ, Handford S, Kaczmarski EB, Edwards-Jones V, Fox AJ (2005) Multilocus sequence typing of Neisseria meningitidis directly from clinical samples and application of the method to the investigation of meningococcal disease case clusters. J Clin Microbiol 43:6007–6014CrossRefPubMedPubMedCentral
18.
go back to reference Diggle MA, Bell CM, Clarke SC (2003) Nucleotide sequence-based typing of meningococci directly from clinical samples. J Med Microbiol 52:505–508CrossRefPubMed Diggle MA, Bell CM, Clarke SC (2003) Nucleotide sequence-based typing of meningococci directly from clinical samples. J Med Microbiol 52:505–508CrossRefPubMed
19.
go back to reference Diggle MA, Clarke SC (2003) Detection and genotyping of meningococci using a nested PCR approach. J Med Microbiol 52:51–57CrossRefPubMed Diggle MA, Clarke SC (2003) Detection and genotyping of meningococci using a nested PCR approach. J Med Microbiol 52:51–57CrossRefPubMed
20.
go back to reference Stefanelli P, Fazio C, Mastrantonio P (2001) Typing of Neisseria meningitidis isolates from patients with invasive disease by molecular analysis of porin genes. New Microbiol 24:149–155PubMed Stefanelli P, Fazio C, Mastrantonio P (2001) Typing of Neisseria meningitidis isolates from patients with invasive disease by molecular analysis of porin genes. New Microbiol 24:149–155PubMed
21.
go back to reference Clarke SC, Diggle MA, Edwards GF (2001) Automated non-culture-based sequence typing of meningococci from body fluids. Br J Biomed Sci 58:230–234PubMed Clarke SC, Diggle MA, Edwards GF (2001) Automated non-culture-based sequence typing of meningococci from body fluids. Br J Biomed Sci 58:230–234PubMed
Metadata
Title
Detection of Neisseria meningitidis DNA from skin lesion biopsy using real-time PCR: usefulness in the aetiological diagnosis of purpura fulminans
Authors
Pierre Staquet
Ludovic Lemee
Eva Verdier
Guy Bonmarchand
Vincent Laudenbach
Christine Michel
Jean-Francois Lemeland
Stephane Marret
Thierry Blanc
Publication date
01-07-2007
Publisher
Springer Berlin Heidelberg
Published in
Intensive Care Medicine / Issue 7/2007
Print ISSN: 0342-4642
Electronic ISSN: 1432-1238
DOI
https://doi.org/10.1007/s00134-007-0667-2

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