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Biomedical subjects

Didier Raoult

Publications and source records attributed to Didier Raoult.

At least 181 records · Page 10Linked to original sources
PubMed2004

Cardiac valves in patients with Whipple endocarditis: microbiological, molecular, quantitative histologic, and immunohistochemical studies of 5 patients.

The pathological features of Whipple endocarditis, which is caused by Tropheryma whipplei, were histologically evaluated in cardiac valves from 5 patients. We used quantitative image analysis to compare the valvular fibrosis, calcifications, vegetations, inflammation, and vascularization due to Whipple endocarditis with those due to non-Whipple endocarditis and degenerative valves. We also studied the presence of T. whipplei in valves by immunohistochemical analysis, culture, and polymerase chain reaction (PCR). In histologic analysis, Whipple endocarditis was characterized by significant fibrosis, a lack of calcifications, slight inflammation and vascularization, and vegetations of intermediate size. Inflammatory infiltrates consisted mainly of foamy macrophages and lymphocytes. We found that the detection of T. whipplei in cardiac valves, by immunohistochemical analysis, was correlated with the detection of the bacterium by culture and PCR. We report, for the first time, the immunodetection of T. whipplei in a surgically removed arterial embolus. Pathological and immunohistologic analyses may contribute to the diagnosis of Whipple endocarditis.

Adult↗
PubMed2004

Multi-pathogens sequence containing plasmids as positive controls for universal detection of potential agents of bioterrorism.

BACKGROUND: The limited circulation of many of the agents that are likely to be used in a bioterrorism attack precludes the ready availability of positive controls. This means that only specialized laboratories can screen for the presence of these agents by nucleic amplification assays. Calibrated controls are also necessary for quantitative measurements. Primers and probes to be used in both conventional and real-time PCR assays were designed for the detection of agents likely to be used by a bioterrorist. Three plasmids, each of which contains 4 to 6 specific sequences from agents on the CDC Category A and B list (excluding RNA viruses) were constructed. Two plasmids incorporate the sequences of Category A and B agents, respectively. The third plasmid incorporates sequences from Variola major and organisms that cause rash-like illnesses that may be clinically confused with smallpox. An "exogenic sequence", introducing a NotI restriction site was incorporated in the native sequences of the bioterrorism agents inserted in plasmids. The designed molecular system for detection of bioterrorism agents was tested on each of these agents (except Monkeypox virus, Smallpox virus and 2 Burkholderia species for which no native DNA was available) and a collection of 50 isolates of C. burnetii using constructed plasmids as positive controls. RESULTS: Designed primers and probes allowed molecular detection, in either single or multiplex assays, of agent-specific targets with analytical sensitivities of between 1 and 100 DNA copies. The plasmids could be used as positive controls. False-positive results due to contamination by the positive control were easily detected by sequencing and eliminated by digestion with NotI. CONCLUSION: Plasmid A and B can be used as positive controls in molecular assays for the detection of bioterrorism agents in clinical specimens or environmental samples. Plasmid C can be used as a positive control in differentiation of vesicular rashes. It is also possible to avoid or to ensure immediate detection of false positive results due to contamination by positive controls using these plasmids. These plasmids and the corresponding primers and probes are immediately available for all clinical microbiology laboratories provided they have molecular amplification equipment.

Bacteria↗
PubMed2004

Lipopolysaccharide from Coxiella burnetii is involved in bacterial phagocytosis, filamentous actin reorganization, and inflammatory responses through Toll-like receptor 4.

The role of Toll-like receptors (TLRs) in the recognition of extracellular and facultative intracellular bacteria by the innate immune system has been extensively studied, but their role in the recognition of obligate intracellular organisms remains unknown. Coxiella burnetii, the agent of Q fever, is an obligate intracellular bacterium that specifically inhabits monocytes/macrophages. We showed in this study that C. burnetii LPS is involved in the uptake of virulent organisms by macrophages but not in that of avirulent variants. The uptake of virulent organisms was dependent on TLR4 because it was reduced in macrophages from TLR4(-/-) mice. In addition, LPS was responsible for filamentous actin reorganization induced by virulent C. burnetii, which was prevented in TLR4(-/-) macrophages. In contrast, the intracellular fate of C. burnetii was not affected in TLR4(-/-) macrophages, suggesting that TLR4 does not control the maturation of C. burnetii phagosome and the microbicidal activity of macrophages. These results are consistent with in vivo experiments because the pattern of tissue infection and the clearance of C. burnetii were similar in wild-type and TLR4(-/-) mice. We also showed that the number of granulomas was decreased in the liver of infected TLR4(-/-) mice, and the formation of splenic granulomas was only transient. The impaired formation of granulomas was associated with decreased production of IFN-gamma and TNF. Taken together, these results demonstrate that TLR4 controls early events of C. burnetii infection such as macrophage phagocytosis, granuloma formation, and cytokine production.

Actins↗
PubMed2004

Effect of sex on Coxiella burnetii infection: protective role of 17beta-estradiol.

Q fever is a zoonosis caused by Coxiella burnetii and recently has been recognized as a potential agent of bioterrorism. In Q fever, men are symptomatic more often than women, despite equal seroprevalence. We hypothesized that sex hormones play a role in the pathogenesis of C. burnetii infection. When C57/BL6 mice were injected with C. burnetii, bacteria load and granuloma numbers were lower in females than in males. Ovarectomized mice showed increased bacteria load in the spleen and the liver, similar to that found in males. The granuloma number was also increased in ovarectomized mice and reached the levels found in males. Tissue infection and granulomatous response are largely under the control of estrogens: treatment of ovarectomized mice with 17beta-estradiol reduced both bacteria loads and granuloma numbers. These results show that sex hormones control host response to C. burnetii infection and may account for host-dependent clinical presentation of Q fever.

Animals↗
PubMed2004

Measurement of the antibiotic susceptibility of Coxiella burnetii using real time PCR.

The objective of this study was to determine MICs of antibiotics for two reference strains of Coxiella burnetii using real time quantitative PCR. The method was very sensitive and specific and allowed the evaluation of the doubling time of Nine Mile and Q212 strains: 37 and 15 h, respectively. Dose response curves of antibiotics were used to determine MICs. Those of doxycycline, fluoroquinolone compounds and rifampicin were in the range 1-4 mg/l. Telithromycin was the most effective macrolide compound with MICs of 1-2 mg/l. The results confirmed previous reports on the accuracy of this new method for the determination of the antibiotic susceptibility of C. burnetii and could be used for the screening of new drugs.

Animals↗
PubMed2004

Tick-borne rickettsioses in international travellers.

BACKGROUND: Tick-borne rickettsioses are of emerging importance in today's travel medicine but have until recently received little attention. We describe the current knowledge of tick-borne rickettsioses as they relate to international travel, their microbiological diagnosis, treatment, possible prevention, and future prospects. METHODS: Literature-based review and personal observations. RESULTS: During the last decade, some 400 cases of tick-borne rickettsioses have been reported in international travellers, the vast majority being African tick bite fever caused by Rickettsia africae and Mediterranean spotted fever caused by Rickettsia conorii. Only a minority of infected travellers can recall a preceding tick bite. Most patients present with a mild-to-moderately severe flu-like illness typically accompanied by a cutaneous rash and an inoculation eschar at the site of the tick bite, but potentially life-threatening disease with disseminated vaculitis is occasionally seen. Definite microbiological confirmation of tick-borne rickettsioses by isolation or antigen detection is only available at reference laboratories and diagnosis must in most cases rely on clinical and epidemiological data supported by serology. Doxycycline is the recommended treatment for tick-borne rickettsioses and prevention is based on personal protective measures against tick bites when travelling in endemic areas. CONCLUSION: Tick-borne rickettsiosis should be suspected in febrile returnees from endemic areas, especially in cases with skin eruptions. Travellers to endemic areas should be encouraged to use personal protective measures against tick bites.

Humans↗
PubMed2004

The history of epidemic typhus.

Few infectious diseases have influenced human civilization to the same degree as louse-transmitted typhus. Rickettsia prowazekii continues to strikes tens to hundreds of thousands of persons who live with war, famine, crowding, and in squalid conditions associated with social unrest, with mortality rates in excess of 10% to 15%. Historical documents confirm that such devastation has been a continuous feature of human existence to the extent that typhus has been a major determinant in the outcome of many wars, altering human history in its wake-despite incomplete knowledge of its precise origin. In the twenty-first century, circumstances are still conductive for outbreak; the emerging threat of bioterrorism raises justifiable concerns that typhus could affect civilization just as greatly in the future as it has in the past.

Disease Outbreaks↗
PubMed2004

What does the future hold for clinical microbiology?

In the past decade, clinical microbiology laboratories have undergone important changes with the introduction of molecular biology techniques and laboratory automation. In the future, there will be a need for more rapid diagnoses, increased standardization of testing and greater adaptability to cope with new threats from infectious microorganisms, such as agents of bioterrorism and emerging pathogens. The combination of the new tools that are now being developed in research laboratories, the general reorganization of clinical laboratories and improved communication between physicians and clinical microbiologists should lead to profound changes in the way that clinical microbiologists work.

Bacteria↗
PubMed2004

Low seroprevalence of bartonella species in danish elite orienteers.

In the 1990s, studies were conducted to investigate 16 episodes of sudden unexpected cardiac death (SUCD) among Swedish elite orienteers during the period from 1979 to 1992. A case control study revealed that a significantly higher proportion of Swedish elite orienteers were B. elizabethae seropositive compared to controls. The aim of our study, designed as a case-control study, was to determine whether similarly high rates of B. elizabethae seropositivity were present among Danish elite orienteers. Cases were 43 elite orienteers; controls were 159 blood donors and 63 elite indoor sportsmen. All participants were tested for antibodies against B. henselae, B. quintana and B. elizabethae using immunofluorescent antibody tests. Surprisingly, Bartonella antibodies were only detected in sera from 5 persons: B. henselae from 1 elite orienteer, 1 handball player and 1 blood donor. B. elizabethae antibodies were detected in 1 handball player and 1 basketball player. We found no association between elite orienteers and the prevalence of Bartonella antibody positivity. This is in contrast to the Swedish study, and might be explained by the use of different serological methods in the 2 studies; to determine whether it is a true difference, a new study is needed.

Adult↗
PubMed2004

Lack of Bartonella sp. in 167 Ixodes ricinus ticks collected in central Sweden.

Sudden death in Swedish orienteers was demonstrated to be significantly associated with antibodies to Bartonella sp. To test if these antibodies could be related with tick exposure, we searched Bartonella sp. in Ixodes ricinus ticks using specific PCR amplification and culture. No Bartonella sp. was detected in 167 ticks tested.

Animals↗
PubMed2004

Serological and molecular evidence of Rickettsia helvetica in Denmark.

Recent seroepidemiological studies and examinations of Ixodes ricinus ticks in Europe have demonstrated the presence of an emerging tick-borne infection with Rickettsia helvetica. We conducted a serosurvey in 168 Danish patients seropositive for borreliosis reflecting their exposure to I. ricinus ticks. A total of 21 patients (12.5%) had positive antibody titres to R. helvetica including 4 cases of seroconversion. None of the samples were positive for antibodies to Ehrlichia. We conclude that in humans exposed to I. ricinus ticks in Denmark the risk of acquiring rickettsial infection is for the first time demonstrated. In the same region of Denmark we collected 570 I. ricinus ticks from various sources, and examinations by PCR for Rickettsia were performed. Positive reactions were obtained in 23 ticks (4%), and R. helvetica was identified in all 13 of those for which sequencing was performed.

Animals↗
PubMed2004

Mycoplasma pneumoniae and Legionella pneumophila in community-acquired lower respiratory tract infections among hospitalized children: diagnosis by real time PCR.

Mycoplasma pneumoniae and Legionella pneumophila are increasingly recognized as important agents of community-acquired lower respiratory tract infections (LRTI). Mycoplasma pneumoniae has been also recognized as a cause of nosocomial infections. The aim of this study was to investigate the role of real time polymerase chain reaction (PCR) for the rapid diagnosis of these infections among hospitalized children with community-acquired LRTI. During 2001, 65 children were prospectively studied. Microbiological investigation consisted of capillary PCR with a LightCycler for M. pneumoniae and L. pneumophila in induced sputum or throat swab specimens, IgM enzyme immunoassay for M. pneumoniae and immunofluorescence for L. pneumophila in paired sera. Serology testing showed acute M. pneumoniae infection in 18 (27.5%) patients and L. pneumophila in 1 (1.5%). M. pneumoniae was also detected in sputum specimen by capillary PCR in 9 (50%) serologically diagnosed cases, including 4 (22%) with non-diagnostic IgM levels in the acute phase. Capillary PCR and IgM enzyme immunoassay diagnosed together 15 (83%) M. pneumoniae cases in the acute phase. It is concluded that M. pneumoniae is an important cause of LRTI necessitating hospitalization among children in Greece. Capillary PCR in sputum may diagnose M. pneumoniae LRTI in the acute setting and direct therapy and isolation of patients.

Adolescent↗
PubMed2004

Rickettsial infections--a threat to travellers?

PURPOSE OF REVIEW: Recent developments in cell-culture techniques and molecular methods have led to the description of several new rickettsial diseases. An update on these new infections should be of interest to health workers with patients who are international travellers. RECENT FINDINGS: Epidemic typhus was reported last year in the United States when an outbreak of murine typhus was recorded in Hawaii. Among spotted fever group rickettsioses, African tick bite fever is now probably the most common rickettsial infection in Africa with numerous cases also reported in international travellers. For the first time the Astrakhan fever rickettsia has been described outside Europe, in a French patient returning from Chad. Similarly, the first case of Rickettsia sibirica mongolotimonae infection in Africa was reported in 2004. Finally, a newly recognized agent of a spotted fever rickettsiosis, Rickettsia parkeri, has been reported in the United States during 2004. SUMMARY: Because results of serological testing are only presumptive, sophisticated methods are crucial for the diagnosis and description of new rickettsial diseases, especially in atypical cases. Modern diagnostic tools include cross-adsorption assays, Western blot testing, and cell-culture and molecular-biological methods.

Animals↗