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Biomedical subjects

Didier Raoult

Publications and source records attributed to Didier Raoult.

At least 145 records · Page 8Linked to original sources

Tick-borne rickettsioses around the world: emerging diseases challenging old concepts.

During most of the 20th century, the epidemiology of tick-borne rickettsioses could be summarized as the occurrence of a single pathogenic rickettsia on each continent. An element of this paradigm suggested that the many other characterized and noncharacterized rickettsiae isolated from ticks were not pathogenic to humans. In this context, it was considered that relatively few tick-borne rickettsiae caused human disease. This concept was modified extensively from 1984 through 2005 by the identification of at least 11 additional rickettsial species or subspecies that cause tick-borne rickettsioses around the world. Of these agents, seven were initially isolated from ticks, often years or decades before a definitive association with human disease was established. We present here the tick-borne rickettsioses described through 2005 and focus on the epidemiological circumstances that have played a role in the emergence of the newly recognized diseases.

Animals↗

Q fever pneumonia: virulence of Coxiella burnetii pathovars in a murine model of aerosol infection.

Q fever is a worldwide zoonosis caused by Coxiella burnetii, a strictly intracellular bacterium that is a potential bioweapon. Humans usually acquires Q fever after inhalation of dust infected by subclinical animals. We used an aerosol exposure apparatus to challenge immunocompetent (BALB/c) and severe combined immunodeficient (SCID) mice with two different strains (strain Nine Mile and strain Q 212) of C. burnetii at two different inocula. Pathological lesions and dissemination of the bacteria were related to the size of the inoculum. SCID mice showed major pulmonary lesions, whereas similarly infected BALB/c mice were more able to eliminate the bacteria. Pathological differences were found between the strains, with Nine Mile showing more severe histological lesions and quantified spread of bacteria. Our animal model could provide a new tool for the study of acute Q fever pneumonia, the development of Q fever in immunodeficient hosts, and the differentiation of pathogenicity among C. burnetii isolates.

Aerosols↗

PCR detection of bacteria on cardiac valves of patients with treated bacterial endocarditis.

We used broad-range PCR amplification and sequencing to detect and identify bacterial DNA in 156 valves of patients treated for infective endocarditis (IE). Bacterial DNA was found more frequently in patients who underwent valve replacement while on antibiotic treatment for IE (60%) than in patients who had completed antibiotic treatment for IE (37%; P = 0.02). We found specific bacterial DNA in valves removed from 11 of 30 patients who had completed antibiotic treatment for IE. Six had no histological evidence of IE. The presence of DNA was significantly correlated with the presence of histologic lesions (P = 0.001) and with the presence of bacteria detected by Gram staining (P < 0.001). Bartonella and streptococci were detected for much longer after antibiotic treatment by PCR than other species (P = 0.047 and 0.04, respectively), and coagulase-negative staphylococci were detected for much shorter periods (P = 0.02). The finding that bacterial DNA was more likely to be detected in valves of patients with active IE than in patients who had completed antibiotic treatment for IE shows that bacterial DNA is cleared slowly. There was no significant correlation between the duration of antibiotic therapy and the presence of bacterial DNA in valves. Since the persistence of bacterial DNA in valves does not necessarily indicate the persistence of viable bacteria, the detection of bacterial DNA in valves from IE patients should be interpreted with caution, in particular in those patients with a past history of treated IE.

Adult↗

Use of shell-vial cell culture assay for isolation of bacteria from clinical specimens: 13 years of experience.

The shell-vial culture assay is performed routinely in our laboratory. Recently we revisited our experience of using the shell-vial culture assay for the isolation of microorganisms from various clinical samples. Over a 13-year period, we have isolated 580 bacterial strains (5%) from 11,083 clinical samples tested. Over the same period, 285 isolates of rickettsiae, bartonellae, or Coxiella burnetii were cultured from a total of 7,102 samples tested. These isolates include 55 Rickettsia sp. isolates, 95 Coxiella burnetii isolates, and 135 Bartonella sp. isolates. Based on our experience with the growth of fastidious microorganisms, we have used a centrifugation shell-vial technique called JNSP, for "je ne sais pas" ("I don't know [what I am growing]") for the isolation of other microorganisms. A total of 173 isolates were cultured from the 3,861 clinical samples tested using the JNSP method. Of these, 40 isolates had not been grown before on usual axenic medium. These include 2 Staphylococcus aureus isolates, 7 isolates of Streptococcus sp. and related genera, 6 Mycobacterium sp. isolates, 1 Nocardia asteroides isolate, 1 Actinomyces sp. isolate, 1 Brucella melitensis isolate, 2 Francisella tularensis isolates, 1 Mycoplasma pneumoniae isolate, and 1 Legionella pneumophila isolate. Using this protocol, we have also cultured intracellular bacteria such as Chlamydia trachomatis and we have performed the first culture and establishment of Trophyrema whipplei. Applied in our laboratory, the shell-vial culture generally exhibits a low rate of success. However, in some cases, this technique allowed microbial diagnosis when classical agar procedure and PCR were negative.

Animals↗

Comparison between rpoB and 16S rRNA gene sequencing for molecular identification of 168 clinical isolates of Corynebacterium.

Higher proportions (91%) of 168 corynebacterial isolates were positively identified by partial rpoB gene determination than by that based on 16S rRNA gene sequences. This method is thus a simple, molecular-analysis-based method for identification of corynebacteria, but it should be used in conjunction with other tests for definitive identification.

Bacterial Typing Techniques↗

Multispacer typing of Rickettsia prowazekii enabling epidemiological studies of epidemic typhus.

Currently, there is no tool for typing Rickettsia prowazekii, the causative agent of epidemic typhus, currently considered a potential bioterrorism agent, at the strain level. To test if the multispacer typing (MST) method could differentiate strains of R. prowazekii, we amplified and sequenced the 25 most variable intergenic spacers between the R. prowazekii and R. conorii genomes in five strains and 10 body louse amplicons of R. prowazekii from various geographic origins. Two intergenic spacers, i.e., rpmE/tRNA(fMet) and serS/virB4, were variable among tested R. prowazekii isolates and allowed identification of three and two genotypes, respectively. When the genotypes obtained from the two spacers were combined, we identified four different genotypes. MST demonstrated that several R. prowazekii strains circulated in human body lice during an outbreak of epidemic typhus in Burundi. This may help to discriminate between natural and intentional outbreaks. Our study supports the usefulness of MST as a versatile method for rickettsial strain genotyping.

Animals↗

[Diseases produced by Bartonella].

The genus Bartonella is cause of a broad number of emerging and re-emerging infectious diseases. Clinical manifestations depend on the implicated Bartonella sp. and the immunity of the host. Because there is not a universal therapy for this infection, treatment should be chosen individually. Bartonella sp. is responsible of potentially serious clinical pictures (endocarditis, chronic bacteremia, bacillary angiomatosis, Carrion's disease, etc.), so clinical suspicion, a quick diagnosis and an early treatment provide a better resolution.

Anti-Bacterial Agents↗

Naming of Rickettsiae and rickettsial diseases.

Over the last 20 years, advances in molecular techniques have greatly facilitated the identification of the members of the Rickettsiales, and numerous new species and diseases have been described. In this paper, we review taxonomic rules and appropriate approaches to valid naming of rickettsial species and the diseases they cause.

Animals↗

Phylogenic analysis of rickettsial patatin-like protein with conserved phospholipase A2 active sites.

Genome analysis of Rickettsia felis highlighted the presence of three patatin-like protein (PLP) genes (pat1, pat2A, and pat2B), whereas only one PLP gene (pat1) is found in the other sequenced rickettsial genomes. Here, we aligned the rickettsial PLPs with characterized patatins from plants and found that they possess all the conserved amino acid residues identified as important for phospholipase A(2) activity. We also carried out a phylogenic analysis of the rickettsial PLPs together with bacterial and eukaryotic homologs. The phylogeny of the rickettsial Pat1 proteins is in conflict with the currently recognized Rickettsia phylogeny. Possible scenarios that might explain this incongruence are discussed and involve either gene conversion or gene duplication events.

Amino Acid Sequence↗

Phylogenetic study of Rickettsia species using sequences of the autotransporter protein-encoding gene sca2.

The analyses of genome sequences from Rickettsia conorii and R. prowazekii have allowed the identification of five genes encoding autotransporter proteins, including ompA and four genes annotated in the R. prowazekii genome as "surface cell antigen" (sca) genes. Of these, ompA and sca5 (ompB) are known to encode membrane-exposed antigenic proteins playing a major role in the host's immune response, and sca4 encodes a truncated autotransporter protein. In order to study further the phylogeny of the genus Rickettsia, we attempted amplification and sequencing of the sca2 genes from the 20 currently validated Rickettsia species. Sixteen species exhibited a complete sca2 gene, ranging from 2,727 bp in R. bellii to 5,580 bp in R. rhipicephali. R. helvetica and R. canadensis had a split gene, and R. prowazekii and R. typhi had remnant fragments of sca2. We also identified in R. akari, R. prowazekii, and R. typhi a duplication of the sca2 gene. The phylogenetic trees inferred from both the nucleotide and protein sequences of sca2 showed four clusters of rickettsiae, that is, the R. rickettsii, R. massiliae, R. akari, and typhus groups, which were supported by significant bootstrap values.

Antigens, Surface↗

TLR2 is necessary to inflammatory response in Coxiella burnetii infection.

The resolution of Q fever, a zoonosis caused by Coxiella burnetii, depends on efficient innate and adaptive immune responses. Such responses are influenced by Toll-like receptors (TLRs). TLR4 is involved only in part in immune responses against C. burnetii, suggesting a role for TLR2. We investigated C. burnetii infection in wild-type and TLR2(-/-) mice. C. burnetii organisms were similarly eliminated by wild-type and TLR2(-/-) mice. In contrast, the formation of granulomas, a marker for efficient cell-mediated immunity, was markedly impaired. These results show that TLR2 is required for inflammatory and immune response to C. burnetii, but is dispensable for bacterial clearance.

Animals↗

Activity of telithromycin against thirteen new isolates of C. burnetii including three resistant to doxycycline.

In this study we have evaluated the in vitro activity of antibiotics against 13 new isolates of Coxiella burnetii using a real-time quantitative PCR assay. MICs against doxycycline ranged from 1 to 8 microg/mL, telithromycin from 0.5 to 2 microg/mL, and all strains had MICs > or = 8 microg/mL for erythromycin. We report that strains resistant to doxycycline exist either in humans or animals.

Animals↗

Molecular screening of Bartonella species in rodents from the Russian Far East.

A potentially broad-range PCR system for gltA gene amplification has been designed. A nearly full (992 bp) sequence of the gltA gene was amplified in two steps. DNA was extracted from organs (spleen and liver) of four mostly spread species of rodents: Apodemus agrarius and Apodemus peninsulae mice; Clethrionomys rufocanus and Microtus fortis voles. All amplicons have been sequenced. The incidence of Bartonella species in rodents was 73%, 60%, 60%, and 83% for the above-mentioned rodents, respectively. Several genotypes have been identified, including two gltA genotypes close to B. taylorii, four gltA genotypes close to B. grahamii, and one genotype of a potentially new species. Several Bartonella species were found in both mice species, although each vole species harbored a single species of Bartonella. High incidence of Bartonella infection was found in all studied species of small rodents in the Russian Far East. Several amplified genotypes probably belong to local strains of B. taylorii and B. grahamii, and one genotype represents a possible new species.

Animals↗

Bacterial zoonoses and infective endocarditis, Algeria.

Blood culture-negative endocarditis is common in Algeria. We describe the etiology of infective endocarditis in this country. Samples from 110 cases in 108 patients were collected in Algiers. Blood cultures were performed in Algeria. Serologic and molecular analysis of valves was performed in France. Infective endocarditis was classified as definite in 77 cases and possible in 33. Causative agents were detected by blood cultures in 48 cases. All 62 blood culture-negative endocarditis cases were tested by serologic or molecular methods or both. Of these, 34 tested negative and 28 had an etiologic agent identified. A total of 18 infective endocarditis cases were caused by zoonotic and arthropodborne bacteria, including Bartonella quintana (14 cases), Brucella melitensis (2 cases), and Coxiella burnetii (2 cases). Our data underline the high prevalence of infective endocarditis caused by Bartonella quintana in northern Africa and the role of serologic and molecular tools for the diagnosis of blood culture-negative endocarditis.

Adolescent↗