Development of a duplex real-time PCR for the detection of Rickettsia spp. and typhus group rickettsia in clinical samples
| Main Authors: | Giulieri, Stefano, Jaton, Katia, Cometta, Alain, Trellu, Laurence T, Greub, Gilbert |
|---|---|
| Format: | Article Journal |
| Terbitan: |
, 2011
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| Subjects: | |
| Online Access: |
https://zenodo.org/record/161899 |
| ctrlnum |
161899 |
|---|---|
| fullrecord |
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<dc schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/ http://www.openarchives.org/OAI/2.0/oai_dc.xsd"><creator>Giulieri, Stefano</creator><creator>Jaton, Katia</creator><creator>Cometta, Alain</creator><creator>Trellu, Laurence T</creator><creator>Greub, Gilbert</creator><date>2011-12-12</date><description>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.</description><identifier>https://zenodo.org/record/161899</identifier><identifier>10.1111/j.1574-695X.2011.00910.x</identifier><identifier>oai:zenodo.org:161899</identifier><relation>pmid:22098502</relation><relation>url:https://zenodo.org/communities/fbm_chuv</relation><rights>info:eu-repo/semantics/openAccess</rights><rights>https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode</rights><source>Fems Immunology and Medical Microbiology 64(1) 92-97</source><subject>Bacterial Proteins/genetics</subject><subject>Citrate (si)-Synthase/genetics</subject><subject>DNA Primers/genetics</subject><subject>Molecular Diagnostic Techniques/methods</subject><subject>Multiplex Polymerase Chain Reaction/methods</subject><subject>Oligonucleotide Probes/genetics</subject><subject>RNA, Bacterial/genetics</subject><subject>RNA, Ribosomal, 16S/genetics</subject><subject>Real-Time Polymerase Chain Reaction/methods</subject><subject>Rickettsia/isolation & purification</subject><subject>Rickettsia Infections/diagnosis</subject><title>Development of a duplex real-time PCR for the detection of Rickettsia spp. and typhus group rickettsia in clinical samples</title><type>Journal:Article</type><type>Journal:Article</type><recordID>161899</recordID></dc>
|
| format |
Journal:Article Journal Journal:Journal |
| author |
Giulieri, Stefano Jaton, Katia Cometta, Alain Trellu, Laurence T Greub, Gilbert |
| title |
Development of a duplex real-time PCR for the detection of Rickettsia spp. and typhus group rickettsia in clinical samples |
| publishDate |
2011 |
| topic |
Bacterial Proteins genetics Citrate (si)-Synthase DNA Primers Molecular Diagnostic Techniques methods Multiplex Polymerase Chain Reaction Oligonucleotide Probes RNA Bacterial/genetics Ribosomal 16S/genetics Real-Time Polymerase Chain Reaction Rickettsia isolation & purification Rickettsia Infections diagnosis |
| url |
https://zenodo.org/record/161899 |
| contents |
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. |
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IOS16997.161899 |
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ZAIN Publications |
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7213 |
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library:special library |
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Cognizance Journal of Multidisciplinary Studies |
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5267 |
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Cognizance Journal of Multidisciplinary Studies |
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16997 |
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Multidisciplinary |
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Stockholm |
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INTERNASIONAL |
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1 |
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2022-06-06T04:22:34Z |
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2022-06-06T04:22:34Z |
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1734901876086800384 |
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17.607244 |