Development of polymerase chain reaction assays for detection of Listeria monocytogenes in clinical cerebrospinal fluid samples.
Détails
Etat: Public
Version: de l'auteur⸱e
ID Serval
serval:BIB_B62872AD2A5F
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Development of polymerase chain reaction assays for detection of Listeria monocytogenes in clinical cerebrospinal fluid samples.
Périodique
Journal of Clinical Microbiology
ISSN
0095-1137 (Print)
ISSN-L
0095-1137
Statut éditorial
Publié
Date de publication
1992
Volume
30
Numéro
8
Pages
1931-1936
Langue
anglais
Notes
Publication types: Journal Article Publication Status: ppublish
Résumé
In order to improve the diagnosis of Listeria meningitis or meningoencephalitis, especially in patients who have received antibiotics before their cerebrospinal fluid (CSF) has been examined, two assays for the detection of Listeria monocytogenes based on the polymerase chain reaction (PCR) were evaluated. After a standard PCR, the amplified DNA was detected either by a second round of PCR with internal primers followed by gel electrophoresis and ethidium bromide staining (nested PCR) or by dot blot hybridization to an internal digoxigenin-labeled probe (PCR-dot blot). For PCR, two sets of primers within the invasion-associated protein gene (iap gene) were chosen. They allowed for the highly specific detection of all L. monocytogenes reference strains tested (serotypes 1/2a, 1/2b, 1/2c, 3a, 3b, 3c, 4a, 4b, 4c, 4d, and 7). These primers did not detect amplification products from various other gram-positive or gram-negative bacterial DNAs or human DNA. The sensitivities of both assays were assessed on sterile CSF samples that were artificially seeded with serial dilutions of L. monocytogenes serotype 4b cells. By both methods the limit of detection was less than 10 cells in the initial reaction. Since the nested PCR is more prone to contamination because of manipulation of the amplified products, a standard PCR assay followed by dot blot hybridization was applied to 52 CSF samples in a retrospective study. Of 28 CSF samples which were sterile or positive for bacteria other than Listeria species, 24 were PCR negative. In contrast, from 17 patients with culture-proven Listeria meningitis, 14 of 17 initial CSF samples were PCR positive, as were 3 of 7 culture-negative followup CSF samples taken after patients received antibiotics. These results support the usefulness of this approach in the diagnosis of Listeria meningitis, in particular, when antibiotic administration precedes culture of CSF.
Mots-clé
Anti-Bacterial Agents/therapeutic use, Base Sequence, DNA, Bacterial/genetics, Evaluation Studies as Topic, Humans, Listeria monocytogenes/genetics, Listeria monocytogenes/isolation & purification, Meningitis, Listeria/cerebrospinal fluid, Meningitis, Listeria/diagnosis, Molecular Sequence Data, Polymerase Chain Reaction/methods
Pubmed
Web of science
Création de la notice
24/01/2008 21:51
Dernière modification de la notice
20/08/2019 16:24