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Biomedical subjects

Didier Raoult

Publications and source records attributed to Didier Raoult.

At least 289 records · Page 16Linked to original sources
PubMed2003

Culture of Tropheryma whipplei from human samples: a 3-year experience (1999 to 2002).

The culture of Tropheryma whipplei, the bacterium responsible for Whipple's disease, has been established only recently. Our objective is to describe, based on our experience, the culture of T. whipplei in HEL cells detected by immunofluorescence staining. Over 3 years, we received 18 samples for T. whipplei culture from 15 patients with Whipple's disease. Ten duodenal biopsy specimens from 10 patients with digestive symptoms were available. Five cardiac valves and three blood samples from five patients with endocarditis were also available. We correlated the results of culture with the type of sample and the culture procedure. Seven isolates were obtained, and three were subsequently established for more than 4 passages. The mean delay for the primary detection was 30 days. The bacterium was isolated more frequently from sterile specimens (5 of 8) than from duodenal biopsy specimens (2 of 10), but the difference (P = 0.14) was not significant. Decontamination of digestive samples containing colistin, amphotericin B, and cephalotin or ciprofloxacin did not impair the isolation of T. whipplei. The use of vancomycin precludes the primary isolation (7 of 12 versus 0 of 6; P = 0.08) and the establishment of T. whipplei (3 of 12 versus 0 of 6; P = 0.5). Omitting samples cultured with vancomycin, the establishment of the strain was significantly higher when antibiotics were prescribed for no more than 7 days (3 of 4 versus 0 of 8; P = 0.03). Our results demonstrate that samples must be collected within 1 week of an antibiotic regimen's initiation for the successful establishment of the bacterium.

Actinomycetales↗
PubMed2003

Bartonella henselae in Ixodes ricinus ticks (Acari: Ixodida) removed from humans, Belluno province, Italy.

The potential role of ticks as vectors of Bartonella species has recently been suggested. In this study, we investigated the presence of Bartonella species in 271 ticks removed from humans in Belluno Province, Italy. By using primers derived from the 60-kDa heat shock protein gene sequences, Bartonella DNA was amplified and sequenced from four Ixodes ricinus ticks (1.48%). To confirm this finding, we performed amplification and partial sequencing of the pap31 protein and the cell division protein ftsZ encoding genes. This process allowed us to definitively identify B. henselae (genotype Houston-1) DNA in the four ticks. Detection of B. henselae in these ticks might represent a highly sensitive form of xenodiagnosis. B. henselae is the first human-infecting Bartonella identified from Ixodes ricinus, a common European tick and the vector of various tickborne pathogens. The role of ticks in the transmission of bartonellosis should be further investigated.

Animals↗
PubMed2003

Molecular detection of Bartonella quintana, B. koehlerae, B. henselae, B. clarridgeiae, Rickettsia felis, and Wolbachia pipientis in cat fleas, France.

The prevalences of Bartonella, Rickettsia, and Wolbachia were investigated in 309 cat fleas from France by polymerase chain reaction (PCR) assay and sequencing with primers derived from the gltA gene for Rickettsia, the its and pap31 genes for Bartonella, and the 16S rRNA gene for Anaplasmataceae. Positive PCR results were confirmed by using the Lightcycler and specific primers for the rOmpB of Rickettsia and gltA of Bartonella. R. felis was detected in 25 fleas (8.1%), W. pipientis, an insect symbiont, in 55 (17.8%), and Bartonella in 81 (26.2%), including B. henselae (9/81; 11.1%), B. clarridgeiae (55/81; 67.9%), B. quintana (14/81; 17.3%), and B. koehlerae (3/81; 3.7%). This is the first report of the amplification of B. quintana from fleas and the first description of B. koehlerae in fleas from an area outside the United States. Cat fleas may be more important vectors of human diseases than previously reported.

Animals↗
PubMed2003

Emerging rickettsioses of the Thai-Myanmar border.

To investigate the presence of rickettsioses in rural residents of the central Thai-Myanmar border, we tested the blood of 46 patients with fever. Four patients had murine typhus, three patients had scrub typhus, and eight patients had spotted fever group rickettsioses, including the first case of Rickettsia felis infection reported in Asia.

Adult↗
PubMed2003

Amoeba-resisting bacteria and ventilator-associated pneumonia.

To evaluate the role of amoeba-associated bacteria as agents of ventilator-associated pneumonia (VAP), we tested the water from an intensive care unit (ICU) every week for 6 months for such bacteria isolates; serum samples and bronchoalveolar lavage samples (BAL) were also obtained from 30 ICU patients. BAL samples were examined for amoeba-associated bacteria DNA by suicide-polymerase chain reaction, and serum samples were tested against ICU amoeba-associated bacteria. A total of 310 amoeba-associated bacteria from 10 species were isolated. Twelve of 30 serum samples seroconverted to one amoeba-associated bacterium isolated in the ICU, mainly Legionella anisa and Bosea massiliensis, the most common isolates from water (p=0.021). Amoeba-associated bacteria DNA was detected in BAL samples from two patients whose samples later seroconverted. Seroconversion was significantly associated with VAP and systemic inflammatory response syndrome, especially in patients for whom no etiologic agent was found by usual microbiologic investigations. Amoeba-associated bacteria might be a cause of VAP in ICUs, especially when microbiologic investigations are negative.

Adult↗
PubMed2003

A survey for spotted fever group rickettsiae and ehrlichiae in Amblyomma variegatum from St. Kitts and Nevis.

Eighty-nine Amblyomma variegatum ticks were collected from the islands of St. Kitts and Nevis in the Caribbean and preserved in 70% ethanol or local rum. After being washed in sterile water, their DNA was extracted and analyzed by a polymerase chain reaction (PCR) for DNA of spotted fever group rickettsiae and ehrlichiae. None of the tested ticks was positive in a PCR assay using the primers 16S EHRD and 16S EHRR for the 16S rRNA gene of Ehrlichia spp.. Forty-one percent of the A. variegatum (36 of 89 of which 34 [47%] of 72 were adult males, 2 (13%) of 16 were adult females, and 0 (0%) of 1 were nymphs) were positive in a PCR assay using the primer pair 190-70 and 190-701 for the outer membrane protein A (ompA) gene of spotted fever group rickettsiae. All PCR amplification products obtained had 100% sequence homology with Rickettsia africae, the agent of African tick-bite fever.

Animals↗
PubMed2003

First isolation of Rickettsia conorii from human blood in Croatia.

AIM: To detect and isolate rickettsial strains from blood samples of patients with presumptive diagnosis of Mediterranean spotted fever (MSF) in the coastal region of south Croatia, and to compare the results with routine serology. METHODS: A "suicide" polymerase chain reaction (PCR), and a shell vial culture were done on samples of ethylenediamine tetra-acetic acid (EDTA) and citrate-anticoagulated blood samples. Indirect immunofluorescence was performed on sera collected from 17 patients clinically diagnosed with MSF during summer in three consecutive years, from 1998 to 2000. RESULTS: The primers used in PCR amplified the expected part of the rickettsia genomic DNA and Rickettsia conorii grew from the shell vial-cultured blood of a single patient. In 13 patients, the diagnosis was confirmed serologically by paired sera, whereas in 4 patients the diagnosis remained presumptive, since no paired sera were available. Analyzing sequences of the ompA and citrate synthase gene, respectively, derived from the shell vial isolate, a 100% similarity with Rickettsia conorii, strain Seven (Malish), was found. CONCLUSION: To the best of our knowledge, this is the first isolation of Rickettsia conorii from a human sample in Croatia, and the first proof of a causative agent of MSF in the country. Beside PCR-based methods and isolation, correct diagnosis of MSF could be still routinely reached by serology.

Base Sequence↗
PubMed2002

IFN-gamma-induced apoptosis and microbicidal activity in monocytes harboring the intracellular bacterium Coxiella burnetii require membrane TNF and homotypic cell adherence.

IFN-gamma is critical for the protection against intracellular bacteria through activation of the antimicrobial machinery of phagocytes. Coxiella burnetii, the etiological agent of Q fever, is a strictly intracellular bacterium that inhabits monocytes/macrophages. We previously showed that IFN-gamma induced C. burnetii killing by promoting the apoptosis of infected monocytes. We show in this study that IFN-gamma-induced apoptosis of infected monocytes was characterized by a time- and dose-dependent activation of caspase-3. IFN-gamma-mediated caspase-3 activation and C. burnetii killing depend on the expression of membrane TNF. Indeed, TNF was transiently expressed on the cell surface of infected monocytes a few hours after IFN-gamma treatment. In addition, anti-TNF Abs inhibited IFN-gamma-mediated caspase-3 activation whereas soluble TNF had no effect on infected cells. Concomitantly, IFN-gamma induced homotypic adherence of C. burnetii-infected monocytes. The latter required the interaction of beta(2) integrins with CD54. When adherence was disrupted by pipetting, by a combination of Abs specific for CD11b, CD18, and CD54, or by an antisense oligonucleotide targeting CD18 mRNA, both cell apoptosis and bacterial killing induced by IFN-gamma were inhibited. Thus, adherence via CD54/beta(2) integrins together with membrane TNF are required to eliminate C. burnetii-infected cells through cell contact-dependent apoptosis. Our results reveal a new component of the antimicrobial arsenal mobilized by IFN-gamma against infection by intracellular bacteria.

Apoptosis↗
PubMed2002

An experimental model of human body louse infection with Rickettsia prowazekii.

Rickettsia prowazekii is transmitted to humans by the body louse. A new experimental model of body louse infection with R. prowazekii is reported here. Eight hundred human lice were infected by feeding on a rabbit that was made bacteremic by injecting 2x106 plaque-forming units of R. prowazekii. The bacterium invaded the stomach cells and was released in feces, in which it was detected 5 days after infection. At day 6 after infection, as a result of the cell burst and the spread of erythrocytes in the hemolymph, the louse became bright red and died within 4 h. The life span of infected lice was shortened by 20-23 days, compared with that of uninfected control lice. Infected lice did not transmit R. prowazekii to their progeny. Through cell culture, rickettsiae were cultivated from fecal samples up to 10 days after their emission. The administration of doxycycline to the rabbit during louse feeding did not cure lice from R. prowazekii infection.

Animals↗
PubMed2002

Coxiella burnetii survival in THP-1 monocytes involves the impairment of phagosome maturation: IFN-gamma mediates its restoration and bacterial killing.

The subversion of microbicidal functions of macrophages by intracellular pathogens is critical for their survival and pathogenicity. The replication of Coxiella burnetii, the agent of Q fever, in acidic phagolysosomes of nonphagocytic cells has been considered as a paradigm of intracellular life of bacteria. We show in this study that C. burnetii survival in THP-1 monocytes was not related to phagosomal pH because bacterial vacuoles were acidic independently of C. burnetii virulence. In contrast, virulent C. burnetii escapes killing in resting THP-1 cells by preventing phagosome maturation. Indeed, C. burnetii vacuoles did not fuse with lysosomes because they were devoid of cathepsin D, and did not accumulate lysosomal trackers; the acquisition of markers of late endosomes and late endosomes-early lysosomes was conserved. In contrast, avirulent variants of C. burnetii were eliminated by monocytes and their vacuoles accumulated late endosomal and lysosomal markers. The fate of virulent C. burnetii in THP-1 monocytes depends on cell activation. Monocyte activation by IFN-gamma restored C. burnetii killing and phagosome maturation as assessed by colocalization of C. burnetii with active cathepsin D. In addition, when IFN-gamma was added before cell infection, it was able to stimulate C. burnetii killing but it also induced vacuolar alkalinization. These findings suggest that IFN-gamma mediates C. burnetii killing via two distinct mechanisms, phagosome maturation, and phagosome alkalinization. Thus, the tuning of vacuole biogenesis is likely a key part of C. burnetii survival and the pathophysiology of Q fever.

Blood Bactericidal Activity↗
PubMed2002

Bartonella quintana in human erythrocytes.

Bartonella quintana is transmitted by body lice among homeless people. Infection with this organism leads to chronic bacteraemia with few symptoms. We looked for B quintana in erythrocytes in a population of homeless people in Marseille, France. In this report we show the intraerythrocytic presence of B quintana in people with symptomless bacteraemia, by use of a specific monoclonal antibody and laser confocal microscopy. Presence of at least 5 x 10(4) colony-forming units per mL was predictive of intracellular infection. However, such infection was not associated with evidence of haemolysis. Colonisation of erythrocytes could allow efficient transmission of B quintana by body lice.

Bacteremia↗
PubMed2002

Spotless rickettsiosis caused by Rickettsia slovaca and associated with Dermacentor ticks.

The pathogenic role of Rickettsia slovaca was first demonstrated in 1997 in a patient who presented with a single inoculation lesion of the scalp and enlarged cervical lymph nodes after receiving a bite from a Dermacentor tick. Subsequently, we evaluated the occurrence of R. slovaca infections among patients living in France and Hungary who presented with these symptoms. R. slovaca infections were confirmed by polymerase chain reaction (PCR) in 17 of 67 enrolled patients. Infections were most likely to occur in patients aged <10 years and in patients who were bitten during the colder months of the year. The median duration of incubation for the disease was 7 days. Fever was present in only 2 patients, and only 1 patient developed a rash. Sequelae included persistent asthenia (3 cases) and localized alopecia (4 cases). Immunofluorescence and/or Western blot analysis detected antibodies in 50% of tested patients. Three Dermacentor ticks obtained from patients revealed R. slovaca by PCR.

Animals↗
PubMed2002

Neurological involvement in acute Q fever: a report of 29 cases and review of the literature.

BACKGROUND: Q fever is characterized by its clinical polymorphism; neurological involvement has occasionally been described. In the course of acute Q fever, neurological manifestations may include aseptic meningitis, encephalitis or encephalomyelitis, and peripheral neuropathy. OBJECTIVE: To review and evaluate cases of acute Q fever with neurological symptoms diagnosed in our laboratory. METHODS: A total of 1269 acute Q fever cases were recorded from January 1985 to January 2000 in our laboratory and were reviewed for neurological complications. Patients were considered to have acute Q fever when serological procedures showed Coxiella burnetii phase II titers of 1:200 or higher for IgG and 1:50 or higher for IgM. Those patients who underwent a lumbar puncture for cerebrospinal fluid analysis or who had abnormal neurological symptoms were selected for this study. We describe the clinical, epidemiological, and biological features of these cases. We also review the literature and compare our cases with those previously reported. RESULTS: Among the 45 patients selected, 14 were excluded because they had normal cerebrospinal fluid and no neurological symptoms. Two were excluded because there were no clinical or epidemiological data. Three major clinical syndromes were observed: meningoencephalitis or encephalitis in 17 cases; meningitis in 8; and myelitis and peripheral neuropathy in 4. Encephalitic signs were not specific, but behavior or psychiatric disturbances were common. CONCLUSIONS: Q fever should be included in the differential diagnosis of acute neurological disease in a patient with a fever. Serological testing should be performed in cases of meningoencephalitis, lymphocytic meningitis, and peripheral neuropathy, including Guillain-Barré syndrome and myelitis.

Acute Disease↗