Development of a duplex real-time PCR for the detection of Rickettsia spp. and typhus group rickettsia in clinical samples.

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Serval ID
serval:BIB_EC516FB23F50
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Development of a duplex real-time PCR for the detection of Rickettsia spp. and typhus group rickettsia in clinical samples.
Journal
FEMS Immunology and Medical Microbiology
Author(s)
Giulieri S., Jaton K., Cometta A., Trellu L.T., Greub G.
ISSN
1574-695X (Electronic)
ISSN-L
0928-8244
Publication state
Published
Issued date
2012
Peer-reviewed
Oui
Volume
64
Number
1
Pages
92-97
Language
english
Abstract
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.
Keywords
Adult, Bacterial Proteins/genetics, Citrate (si)-Synthase/genetics, DNA Primers/genetics, Female, Humans, Male, Molecular Diagnostic Techniques/methods, Multiplex Polymerase Chain Reaction/methods, Oligonucleotide Probes/genetics, RNA, Bacterial/genetics, RNA, Ribosomal, 16S/genetics, Real-Time Polymerase Chain Reaction/methods, Rickettsia/isolation & purification, Rickettsia Infections/diagnosis, Sensitivity and Specificity
Pubmed
Web of science
Open Access
Yes
Create date
28/02/2012 15:24
Last modification date
20/08/2019 16:14
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