Coxiella burnetii vascular graft infection

Senn, Laurence; Franciolli, Mario; Raoult, Didier; Moulin, Alexandre; Segesser, Ludwig von; Calandra, Thierry; Greub, Gilbert

doi:10.1186/1471-2334-5-109

Cited by 25 publications

(15 citation statements)

References 16 publications

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“…During 7 years of clinical use, our PCR was positive on various non‐respiratory samples such as cardiac valve, aortic aneurysm, liver biopsy and vertebral biopsy (Table ), congruent with manifestations of Q fever already reported in the literature . All these positive results were also positive by serology, confirming the excellent specificity of our test over a 7‐year period.…”

Section: Discussionsupporting

confidence: 85%

“…Early diagnosis of Q fever is important because delayed diagnosis may lead to severe chronic infections such as endocarditis and endovascular infections with high morbidity and mortality . As a matter of fact, despite the poor performance of PCR on serum , sera are often used instead of EDTA‐blood because serum is often already available for serology.…”

Section: Discussionmentioning

confidence: 99%

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Development of a high throughput PCR to detect Coxiella burnetii and its application in a diagnostic laboratory over a 7-year period

Jaton

Péter

Raoult

et al. 2013

New Microbes and New Infections

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Q fever is a worldwide zoonotic infectious disease due to Coxiella burnetii . The clinical presentation may be acute (pneumonia and/or hepatitis) or chronic (most commonly endocarditis). Diagnosis mainly relies on serology and PCR. We therefore developed a quantitative real-time PCR. We first tested blindly its performance on various clinical samples and then, when thoroughly validated, we applied it during a 7-year period for the diagnosis of both acute and persistent C. burnetii infection. Analytical sensitivity (< 10 copies/PCR) was excellent. When tested blindly on 183 samples, the specificity of the PCR was 100% (142/142) and the sensitivity was 71% (29/41). The sensitivity was 88% (7/8) on valvular samples, 69% (20/29) on blood samples and 50% (2/4) on urine samples. This new quantitative PCR was then successfully applied for the diagnosis of acute Q fever and endovascular infection due to C. burnetii , allowing the diagnosis of Q fever in six patients over a 7-year period. During a local small cluster of cases, the PCR was also applied to blood from 1355 blood donors; all were negative confirming the high specificity of this test. In conclusion, we developed a highly specific method with excellent sensitivity, which may be used on sera for the diagnosis of acute Q fever and on various samples such as sera, valvular samples, aortic specimens, bone and liver, for the diagnosis of persistent C. burnetii infection.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Development of a high throughput PCR to detect Coxiella burnetii and its application in a diagnostic laboratory over a 7-year period

Jaton

Péter

Raoult

et al. 2013

New Microbes and New Infections

View full text Add to dashboard Cite

show abstract

“…It has been shown to be useful in the microbiological diagnosis of brain abscesses (16), endocarditis (13), infected prosthetic joints (20), meningitis (2,5,18), osteomyelitis (9,21), septic arthritis (17), spondylodiscitis (8), and vascular graft infections (19). In our material, samples from these conditions comprised 27% (70 samples) of the total, and 85% of those positive by the broad-range PCR produced pure chromatograms.…”

Section: Resultsmentioning

confidence: 99%

Direct 16S rRNA Gene Sequencing from Clinical Specimens, with Special Focus on Polybacterial Samples and Interpretation of Mixed DNA Chromatograms

et al. 2009

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RipSeq (iSentio, Bergen, Norway) is a web-based application for the analysis of mixed DNA chromatograms.It opens the possibility to analyze chromatograms obtained by direct 16S rRNA gene sequencing from polybacterial human clinical samples. In this study, we used direct 16S rRNA gene sequencing to investigate 264 samples from a wide range of suspected human bacterial infections. The sequence-based identification was compared with the results from routine culture-based identification. A total of 151 samples were positive by the first PCR, producing 85 pure and 66 mixed DNA chromatograms. All mixed chromatograms were analyzed by RipSeq, although seven were so complex that only the dominant bacterial sequences could be identified. In general, sequence-based identification detected a larger number of species than did culture for samples from patients who had received antibiotics prior to sample collection and for samples containing anaerobic bacteria. RipSeq made it possible to apply this supplementary diagnostic tool to typical polybacterial specimens, such as internal abscesses, pleural fluids, and bile.Detection and identification of bacteria directly from clinical samples by broad-range PCR targeting the 16S rRNA gene and DNA sequencing (direct 16S rRNA gene sequencing) make up a well-established method in many laboratories. This method gives the possibility to identify bacteria that died during transportation or as a consequence of antibiotic treatment and to uncover bacteria with special growth requirements. The latest advances in PCR and sequencing technology also offer a more rapid identification than that obtained with standard phenotypic methods that depend on bacterial growth.Because of difficulties in the interpretation of DNA chromatograms resulting from direct sequencing of polybacterial samples, the use of this diagnostic tool has been limited to infections that are predominantly monobacterial. We earlier described RipSeq (iSentio, Bergen, Norway), a web-based application for the analysis of mixed DNA chromatograms (12). In the same article, we presented the RipSeq performance on a number of mixed DNA chromatograms obtained by direct sequencing from saline suspensions containing two and three different bacterial species and discussed the possible benefits and limitations one could experience if the method was to be applied to human clinical samples. In this study, direct 16S rRNA gene sequencing was used to investigate 264 human clinical samples from a wide range of locations, including typical polybacterial specimens such as abscesses and pleural fluids. All mixed DNA chromatograms were analyzed with the RipSeq program, and the sequence-based results were compared to routine culture-based diagnostics in our hospital laboratory.We also discuss special concerns in the selection of a lysis procedure and primers for use on polymicrobial samples as well as the establishment of a reliable negative control.

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“…It is clear from this and other studies that the results achieved by routine culture-based diagnostics alone are not sufficient when it comes to patients who have received one or more doses of antibiotics prior to sample collection, or the investigation of anaerobe infections (Al Masalma et al , 2009; Goldenberger et al , 1997; Kommedal et al , 2009; Petti et al , 2008b; Schuurman et al , 2004; Senn et al , 2005). Infectious-disease physicians are well aware of these limitations.…”

Section: Discussionmentioning

confidence: 99%

Characterization of polybacterial clinical samples using a set of group-specific broad-range primers targeting the 16S rRNA gene followed by DNA sequencing and RipSeq analysis

Kommedal

Lekang

Langeland

et al. 2011

Journal of Medical Microbiology

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The standard use of a single universal broad-range PCR in direct 16S rDNA sequencing from polybacterial samples leaves the minor constituents at risk of remaining undetected because all bacterial DNA will be competing for the same reagents. In this article we introduce a set of three broad-range group-specific 16S rDNA PCRs that together cover the clinically relevant bacteria and apply them in the investigation of 25 polybacterial clinical samples. Mixed DNA chromatograms from samples containing more than one species per primer group were analysed using RipSeq Mixed (iSentio, Norway), a web-based application for the interpretation of chromatograms containing up to three different species. The group-specific PCRs reduced complexity in the resulting DNA chromatograms and made the assay more sensitive in situations with unequal species concentrations. Together this allowed for identification of a significantly higher number of bacterial species than did standard direct sequencing with a single universal primer pair and RipSeq analysis (95 vs 51). The method could improve microbiological diagnostics for important groups of patients and can be established in any laboratory with experience in direct 16S rDNA sequencing.

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Coxiella burnetii vascular graft infection

Abstract: Background: Coxiella burnetii, the causative agent of Q fever, may cause culture-negative vascular graft infections. Very few cases of C. burnetii infection of a vascular graft have been reported. All were diagnosed by serology.

Cited by 25 publications

References 16 publications

Development of a high throughput PCR to detect Coxiella burnetii and its application in a diagnostic laboratory over a 7-year period

Development of a high throughput PCR to detect Coxiella burnetii and its application in a diagnostic laboratory over a 7-year period

Direct 16S rRNA Gene Sequencing from Clinical Specimens, with Special Focus on Polybacterial Samples and Interpretation of Mixed DNA Chromatograms

Characterization of polybacterial clinical samples using a set of group-specific broad-range primers targeting the 16S rRNA gene followed by DNA sequencing and RipSeq analysis

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