Titre
Development of a duplex real-time PCR for the detection of Rickettsia spp. and typhus group rickettsia in clinical samples.
Type
article
Institution
UNIL/CHUV/Unisanté + institutions partenaires
Périodique
FEMS Immunology and Medical Microbiology
Auteur(s)
Giulieri, S.
Auteure/Auteur
Jaton, K.
Auteure/Auteur
Cometta, A.
Auteure/Auteur
Trellu, L.T.
Auteure/Auteur
Greub, G.
Auteure/Auteur
Liens vers les personnes
Liens vers les unités
ISSN
1574-695X
Statut éditorial
Publié
Date de publication
2012
Volume
64
Numéro
1
Première page
92
Dernière page/numéro d’article
97
Peer-reviewed
Oui
Langue
anglais
Résumé
Molecular diagnosis using real-time polymerase chain reaction (PCR) may allow earlier diagnosis of rickettsiosis. We developed a duplex real-time PCR that amplifies (1) DNA of any rickettsial species and (2) DNA of both typhus group rickettsia, that is, Rickettsia prowazekii and Rickettsia typhi. Primers and probes were selected to amplify a segment of the 16S rRNA gene of Rickettsia spp. for the pan-rickettsial PCR and the citrate synthase gene (gltA) for the typhus group rickettsia PCR. Analytical sensitivity was 10 copies of control plasmid DNA per reaction. No cross-amplification was observed when testing human DNA and 22 pathogens or skin commensals. Real-time PCR was applied to 16 clinical samples. Rickettsial DNA was detected in the skin biopsies of three patients. In one patient with severe murine typhus, the typhus group PCR was positive in a skin biopsy from a petechial lesion and seroconversion was later documented. The two other patients with negative typhus group PCR suffered from Mediterranean and African spotted fever, respectively; in both cases, skin biopsy was performed on the eschar. Our duplex real-time PCR showed a good analytical sensitivity and specificity, allowing early diagnosis of rickettsiosis among three patients, and recognition of typhus in one of them.
Sujets
PID Serval
serval:BIB_EC516FB23F50
PMID
Open Access
Oui
Date de création
2012-02-28T14:24:27.474Z
Date de création dans IRIS
2025-05-21T05:36:01Z
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Nom
BIB_EC516FB23F50.P001.pdf
Version du manuscrit
preprint
Taille
874.6 KB
Format
Adobe PDF
PID Serval
serval:BIB_EC516FB23F50.P001
URN
urn:nbn:ch:serval-BIB_EC516FB23F500
Somme de contrôle
(MD5):fc911d15c99654384f4ca947c9b7f6ab