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1.
Prog Urol ; 31(12): 699-708, 2021 Oct.
Article in French | MEDLINE | ID: mdl-34154956

ABSTRACT

INTRODUCTION: High-intensity focused ultrasound (HIFU) has proved to be effective in the treatment of localized prostate cancer. The aim of this prospective study is to assess their first oncological and functional results in an Afro-Caribbean population. METHODS: From May 2018 to January 2020, 77 patients issued from French West Indies were included. Several treatments were carried out: whole-gland treatment hemi or focal ablation; in a primary setting (group I) or a salvage therapy (group II). PSA level was assessed at 2, 6, 9 and 12 months. MpMRI and post HIFU biopsy were performed between 6 and 9 months postoperatively. Continence, urinary end erectile functions were assessed by ICS, IPSS and IIEF scores. RESULTS: Groupe I included 71.2% patients, group II, 28.8%. The median age was 75.4 years [IQR 69.6-79.4]. The median follow-up was 8.3 months [IQR 3.5-12.25]. At inclusion, PSA was 7.7ng/ml [IQR 5.5-11.2] in group I, and 5.9ng/ml [IQR 4.4-7.9] in group II. In the whole population, there was 73.5% negative biopsies; 14.7% of the biopsies were positive in treated zone and 11.8% in non-treated zone. Regarding morbidities, urinary incontinence appeared in 7.5% and erectile dysfunction rate was 13.2%. CONCLUSION: Our study reveals the first experience of HIFU by Focal One® device in an Afro-Caribbean population. It seems to be a safe and reproducible treatment with acceptable oncological results and low genitourinary morbidity. Long term follow-up and a higher number of patients are necessary to validate these results.


Subject(s)
Prostatic Neoplasms , Ultrasound, High-Intensity Focused, Transrectal , Aged , Caribbean Region , Humans , Male , Prospective Studies , Prostatic Neoplasms/surgery , Treatment Outcome , Ultrasound, High-Intensity Focused, Transrectal/adverse effects
2.
Prog Urol ; 30(10): 532-540, 2020 Sep.
Article in English | MEDLINE | ID: mdl-32593527

ABSTRACT

BACKGROUND: Prostate cancer is supposedly more aggressive among Afro-Caribbean men. There is a lack of data in this population for active surveillance. Published series are retrospective or have small samples and results are discordant. The objective was to determinate whether actual active surveillance modalities can be applied for Afro-Caribbean men by comparing their oncological outcomes with Caucasian men. METHODS: A total of 449 consecutive patients who underwent active surveillance for favorable-risk prostate cancer in two French University-Medical-Centers between 2005 and 2018: 261 in Guadeloupe, French West Indies, and 188 in Bordeaux, metropolitan France. Median follow-up was 56 months, (95% CI [32-81]) and 52 months (95% CI [30-75]), respectively (P=0.07). Curative treatment was given in case of histological, biological, or imaging progression, or upon patient demand. Primary endpoints were treatment-free, overall and specific survival. Secondary outcomes were reasons of discontinuating active surveillance, histological poor prognosis factors after prostatectomy, CAPRA-S score, biochemical-recurrence-free after treatment and metastasis-free survival. Kaplan-Meier method was used. RESULTS: Median treatment free survival was 58.4 months (CI 95% [48.6-83.1]) for ACM and not reached at 120 months for CM (P=0.002). Overall survival (P=0.53), and specific survival (P=0.21) were similar in the two groups. CM were likely to have poor prognosis factor on prostatecomy piece (57 vs 30%, P=0.01). No difference for repartition of the CAPRA-S score (P=0.86), biochemical-recurrence-free (P=0.92) and metastasis-free (P=0.44) survival. CONCLUSIONS: Oncological outcomes for active surveillance of Afro-Caribbean and Caucasian men were similar in terms of mortality, recurrence and metastasis in our bicentric study, showing usability of current criteria for Afro-Caribbean. The higher rate of disease progression in the Afro-Caribbean population requires close monitoring. LEVEL OF EVIDENCE: 3.


Subject(s)
Black People , Prostatic Neoplasms/therapy , Watchful Waiting , White People , Aged , Caribbean Region , Cohort Studies , France , Guadeloupe , Humans , Male , Middle Aged , Retrospective Studies , West Indies
3.
Prog Urol ; 28(16): 906-914, 2018 Dec.
Article in French | MEDLINE | ID: mdl-30219645

ABSTRACT

INTRODUCTION: Anthropometric data report that pelvic bone of African subjects are narrower and the pelvic cavity is deeper. The aim of the study was to evaluate the influence of pelvic dimensions (PD) on Positive surgical margins (PSM) rate in Afro-Caribbean population after robot-assisted laparoscopic prostatectomy (RALP). PATIENTS AND METHODS: Preoperative pelvic MRI of all patients who have had RALP at the University Hospital Center of Guadeloupe between January 2013 and December 2015 was retrospectively analyzed. PD, including the Height of the upper edge of the prostate (HP), the Apical Depth (AD) and Ischial Spines Distance (ISD), and indexes (prostate volumetric index [ISD/VP], apical depth index [ISD/AD] and prostate depth index [ISD/(AD/HP)]) were compared according to the presence or absence of PSM with uni and multivariate analysis. RESULTS: One hundred and seventy-eight patients were included in the study, of whom 60 (33.7%) presented PSM. In univariate analysis, significant differences between the presence or absence of PSM were observed on the AD (30.3±8.7mm versus 24.8±8.0mm, P<0.001), the HP (9.5±8.5mm versus 16.8±11.9mm, P<0.001) and the ISD (89.6±8.8mm versus 96.1±8.4mm) as well as the indexes of apical depth and prostatic depth. In multivariate logistic regression, the ISD (P<0.001) and HP (P=0.02) were associated with increased likelihood of PSM, but not AD or indexes. CONCLUSION: This study suggests that interspinous distance is the best predictor of PSM during RALP in Afro-Caribbean patients. This measure may be useful to define the therapeutic pattern of patients with prostate cancer. A prospective study with a larger population, comparing RALP in Afro-Caribbean and in caucasians patients, would be needed.


Subject(s)
Body Weights and Measures/methods , Margins of Excision , Pelvis/diagnostic imaging , Prostatectomy , Prostatic Neoplasms/pathology , Prostatic Neoplasms/surgery , Robotic Surgical Procedures , Aged , Body Weights and Measures/standards , Guadeloupe , Humans , Laparoscopy/adverse effects , Laparoscopy/instrumentation , Laparoscopy/methods , Magnetic Resonance Imaging , Male , Middle Aged , Neoplasm Recurrence, Local/diagnosis , Neoplasm Recurrence, Local/pathology , Neoplasm, Residual , Pelvic Bones/pathology , Pelvis/pathology , Preoperative Period , Prognosis , Prostatectomy/adverse effects , Prostatectomy/instrumentation , Prostatectomy/methods , Retrospective Studies , Risk Factors , Robotic Surgical Procedures/adverse effects , Robotic Surgical Procedures/methods
4.
Nat Commun ; 9(1): 2864, 2018 07 20.
Article in English | MEDLINE | ID: mdl-30030428

ABSTRACT

The mechanisms of transfer of crustal material from the subducting slab to the overlying mantle wedge are still debated. Mélange rocks, formed by mixing of sediments, oceanic crust, and ultramafics along the slab-mantle interface, are predicted to ascend as diapirs from the slab-top and transfer their compositional signatures to the source region of arc magmas. However, the compositions of melts that result from the interaction of mélanges with a peridotite wedge remain unknown. Here we present experimental evidence that melting of peridotite hybridized by mélanges produces melts that carry the major and trace element abundances observed in natural arc magmas. We propose that differences in nature and relative contributions of mélanges hybridizing the mantle produce a range of primary arc magmas, from tholeiitic to calc-alkaline. Thus, assimilation of mélanges into the wedge may play a key role in transferring subduction signatures from the slab to the source of arc magmas.

5.
Prog Urol ; 28(2): 114-119, 2018 Feb.
Article in French | MEDLINE | ID: mdl-29162379

ABSTRACT

INTRODUCTION: The incidence of urolithiasis is increasing with dietary changes especially in developed countries. Guadeloupe is a French department overseas where western diet meets traditional local food. The objective was to describe and analyze the epidemiology of urolithiasis in Guadeloupe. MATERIAL AND METHODS: We conducted a retrospective single-center study throughout the year 2015 on patients hospitalized for urolithiasis at University Hospital of Pointe-à-Pitre. Data of the patients, treatments performed and the types of stones were recorded. According to their mineral content, groups were composed. RESULTS: In total, 165 patients were included. The sex ratio was 1.61. The median body mass index (BMI) was 26.5kg/m2. The most common stone was oxalocalcic (64.7%). Mixed stones (24.7%) were in second place. There were only 3.5% of uric acid urolithiasis. Calcium oxalate stones were predominantly monohydrate. The oxalocalcic stones were significantly more frequent in men (80% versus 47.5%, P=0.01) and in the age group over 50 years old (72.2% versus 51.6%, P=0.04). There was no association between the type of stone and the BMI. CONCLUSION: Epidemiology of urolithiasis in our French Caribbean island is, therefore, similar to continental France. However, our population is distinguished by the proportion of women affected and by the different proportions among each type of stone. Other studies on larger samples are needed to study these specificities. LEVEL OF EVIDENCE: 4.


Subject(s)
Urolithiasis/epidemiology , Body Mass Index , Female , Guadeloupe/epidemiology , Humans , Male , Middle Aged , Retrospective Studies
6.
Prog Urol ; 27(8-9): 467-473, 2017.
Article in French | MEDLINE | ID: mdl-28576421

ABSTRACT

PURPOSE: Research of predictive factors of biochemical recurrence to guide the establishment of an adjuvant treatment after radical prostatectomy for cancer with positive surgical margins. METHODOLOGY: A retrospective cohort of 1577 afro-caribbean patients undergoing radical prostatectomy operated between 1st January 2000 and 1st July 2013 was analyzed. In this cohort, 406 patients had positive surgical margin, we excluded 11 patients who received adjuvant therapy (radiotherapy, hormonotherapy, radio-hormonotherapy) and 2 patients for whom histological analysis of the surgical specimen was for a pT4 pathological stage. After a descriptive analysis, we used a Cox model to look for predictors of survival without biochemical recurrence then, depending on the significant variables, we separated our population into six groups: stage pT2 with Gleason score≤3+4 (group 1), stage pT2 with a score of Gleason≥4+3 (group 2), stage pT3a with a Gleason core≤3+4 (group 3), pT3a stage with a score of Gleason≥4+3 (group 4), stage pT3b with a Gleason score≤3+4 (group 5) and stage pT3b Gleason≥with a score of 4+3 (group 6) and compared survival without biochemical recurrence using a log rank test. After radical prostatectomy with surgical margins with an anatomopathological stage≤pT3b, a Gleason score≥4+3 had a pejorative survival without biochemical recurrence than pathological stage (P<0.001). RESULTS: In multivariate analysis, predictors of survival without biochemical recurrence after radical prostatectomy with positive surgical margins were the majority Gleason postoperative (P<0.0001), pathological stage (P=0.049) adjusted preoperative PSA (P=0.826), with the body mass index (BMI) (P=0.59) and tumor volume (P=0.95). CONCLUSION: A high postoperatively Gleason score (≥4+3) has a better predictive value of biochemical recurrence than pathological stage pT2 or pT3 at the patients having been treated for prostate cancer by radical prostatectomy with positive surgical margins. LEVEL OF EVIDENCE: 4.


Subject(s)
Adenocarcinoma/ethnology , Black People/statistics & numerical data , Neoplasm Recurrence, Local/ethnology , Prostatectomy/statistics & numerical data , Prostatic Neoplasms/ethnology , Adenocarcinoma/blood , Adenocarcinoma/pathology , Adenocarcinoma/surgery , Aged , Biomarkers, Tumor/blood , Caribbean Region/epidemiology , France/epidemiology , Humans , Male , Margins of Excision , Middle Aged , Neoplasm Grading , Neoplasm Invasiveness , Neoplasm Recurrence, Local/pathology , Neoplasm Recurrence, Local/surgery , Predictive Value of Tests , Prostate-Specific Antigen/blood , Prostatic Neoplasms/blood , Prostatic Neoplasms/pathology , Prostatic Neoplasms/surgery , Retrospective Studies , Sensitivity and Specificity , Treatment Outcome
7.
J Nutr Health Aging ; 20(3): 347-54, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26892585

ABSTRACT

BACKGROUND: Vascular aging is accompanied by gradual remodeling affecting both arterial and cardiac structure and mechanical properties. Hypertension is suggested to exert pro-inflammatory actions enhancing arterial stiffness. OBJECTIVE: To determine the influence of thoracic aortic inflammation and calcifications on arterial stiffness and cardiac function in hypertensive and normotensive older subjects. DESIGN: A prospective study. SETTING: An acute geriatrics ward of the University Hospital of Nancy in France. SUBJECTS: Thirty individuals ≥ 65 years were examined, including 15 hypertensive subjects and 15 controls well-matched for age and sex. MEASUREMENTS: Applanation tonometry was used to measure aortic pulse wave velocity (AoPWV) and carotid/brachial pulse pressure amplification (PPA). Left ventricular parameters were measured with magnetic resonance imaging. Local thoracic aortic inflammation and calcification were measured by 18 F-fluorodeoxyglucose positron emission tomography/computed tomography imaging. Biomarkers of low-grade inflammation were also quantified. RESULTS: AoPWV was higher in elderly hypertensive subjects comparatively to normotensive controls (15.5±5.3 vs. 11.9±2.5, p=0.046), and hypertensives had a higher calcification volume. In the overall population, calcifications of the thoracic descending aorta and inflammation of the ascending aorta accounted for respectively 18.1% (p=0.01) and 9.6% (p=0.07) of AoPWV variation. Individuals with high levels of calcifications and/or inflammation had higher AoPWV (p=0.003). Inflammation had a negative effect on PPA explaining 13.8% of its variation (p<0.05). CONCLUSION: This study highlights the importance of local ascending aortic inflammation as a potential major actor in the determination of PPA while calcifications and hypertension are more linked to AoPWV. Assessment of PPA in the very elderly could provide complementary information to improve diagnostic and therapeutic strategies targeting ascending aortic inflammation.


Subject(s)
Aorta, Thoracic/pathology , Aorta, Thoracic/physiopathology , Blood Pressure , Calcinosis/physiopathology , Hypertension/physiopathology , Inflammation/physiopathology , Vascular Stiffness , Aged , Aorta/pathology , Aorta/physiopathology , Biomarkers , Calcinosis/complications , Calcinosis/pathology , Female , France , Humans , Hypertension/complications , Inflammation/complications , Inflammation/pathology , Male , Prospective Studies , Pulse Wave Analysis
11.
Eur J Clin Microbiol Infect Dis ; 28(11): 1363-8, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19685089

ABSTRACT

The aim of this study was to compare 16 S rRNA gene amplification and sequencing with a systematic real-time PCR assay screening strategy that includes all common known pathogens recovered from lymph node biopsy specimens. Lymph node biopsy samples sent to our laboratory from January 2007 to December 2008 were tested in the study. Lymph nodes were screened for the presence of any bacteria by PCR amplification and sequencing targeting the 16 S rRNA gene and also by a specific real-time PCR strategy that includes Bartonella henselae, mycobacteria, Francisella tularensis, and Tropheryma whipplei. By testing 491 lymph nodes, we found that the sensitivity of our specific real-time PCR assay strategy was significantly higher than 16 S rRNA PCR amplification and sequencing for the detection of Bartonella henselae (142 vs 98; p < 10(-4)), Francisella tularensis (16 vs 10, p < 10(-4)), and mycobacteria (8 versus 3, p < 10(-4)). None of the samples was positive for Tropheryma whipplei. Our study demonstrates the usefulness and specificity of a systematic real-time PCR strategy for molecular analysis of lymph node biopsy specimens and the higher sensitivity compared with standard 16 S rRNA gene amplification and sequencing.


Subject(s)
Bacteria/isolation & purification , Bacterial Infections/diagnosis , Bacteriological Techniques/methods , Lymphadenitis/microbiology , Polymerase Chain Reaction/methods , Bacteria/genetics , Bacterial Infections/microbiology , Humans , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Sensitivity and Specificity , Sequence Analysis, DNA
13.
Eur J Clin Microbiol Infect Dis ; 28(6): 569-73, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19048317

ABSTRACT

The SeptiFast test (Roche Diagnostics) is a new commercial molecular technique that has emerged for the detection of bacteria in blood. We compared in this study the sensitivity of blood culture to a commercially available broad-range real-time polymerase chain reaction (PCR) assay for the detection in blood of 19 bacterial species and six fungal species (SeptiFast test, Roche Diagnostics) in 63 patients with infectious endocarditis (IE). The SeptiFast test is not more sensitive for organisms such as Streptococci, Enterococci, and Staphylococcus aureus (11/29 versus 12/29 for blood culture). It has detected less commonly coagulase-negative Staphylococci (0/15 versus 3/15, P = 0.2) and significantly fewer other microorganisms (0/6 versus 4/6, P = 0.03). However, bacteria were detected from three IE treated by antibiotics, with blood culture negative on admission. The SeptiFast test may be useful in cases of IE in patients treated with antibiotics before admission.


Subject(s)
Bacteria/isolation & purification , Bacterial Infections/diagnosis , Blood/microbiology , Endocarditis, Bacterial/diagnosis , Fungi/isolation & purification , Molecular Diagnostic Techniques/methods , Mycoses/diagnosis , Bacteria/genetics , Fungi/genetics , Humans , Sensitivity and Specificity
14.
J Mol Endocrinol ; 39(4): 305-18, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17909269

ABSTRACT

The estrogen receptor alpha (ER alpha) status of breast tumors is used to identify patients who may respond to endocrine agents such as tamoxifen. However, ER alpha status alone is not perfectly predictive, and there is a pressing need for more reliable markers of endocrine responsiveness. In this aim, we used a two-step strategy. We first screened genes of interest by a pangenomic 44 K oligonucleotide microarray in a series of ten ER alpha-positive tumors from five tamoxifen-treated postmenopausal patients who relapsed (distant metastasis) and five tamoxifen-treated postmenopausal patients who did not relapse, matched with respect to age, Scarff-Bloom-Richardson grade, lymph node status, and macroscopic tumor size. Genes of interest (n=24) were then investigated in an independent well-characterized series of ER alpha-positive unilateral invasive primary breast tumors from postmenopausal women who received tamoxifen alone as adjuvant hormone therapy after primary surgery. We identified four genes (HRPAP20, TIMELESS, PTPLB, and MGC29814) for which high mRNA levels were significantly associated with shorter relapse-free survival (log-rank test). We also showed that hormone-regulated proliferation-associated 20 kDa protein (HRPAP20) and TIMELESS are 17beta-estradiol-regulated in vitro and are ectopically expressed in OH-Tam-resistant cell lines. In conclusion, these findings point to HRPAP20 and TIMELESS as promising markers of tamoxifen resistance in women with ER alpha-positive breast tumors.


Subject(s)
Breast Neoplasms/drug therapy , Breast Neoplasms/genetics , Calmodulin-Binding Proteins/genetics , Carcinoma/drug therapy , Carcinoma/genetics , Cell Cycle Proteins/genetics , Estrogen Receptor alpha/genetics , Intracellular Signaling Peptides and Proteins/genetics , Oligonucleotide Array Sequence Analysis , Postmenopause , Tamoxifen/therapeutic use , Aged , Antineoplastic Agents, Hormonal/therapeutic use , Biomarkers, Tumor/genetics , Biomarkers, Tumor/physiology , Breast Neoplasms/diagnosis , Calmodulin-Binding Proteins/physiology , Carcinoma/diagnosis , Cell Cycle Proteins/physiology , Disease-Free Survival , Female , Gene Expression Profiling , Gene Expression Regulation, Neoplastic/drug effects , Humans , Intracellular Signaling Peptides and Proteins/physiology , Middle Aged , Postmenopause/genetics , Prognosis , Treatment Outcome , Tumor Cells, Cultured
15.
J Clin Microbiol ; 43(10): 5388-90, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16208026

ABSTRACT

Shewanella spp. are infrequently recovered from clinical specimens. We report here on the first case of osteomyelitis due to Shewanella algae. This bacterium, at first misidentified by phenotypic tests as Shewanella putrefaciens, was subsequently identified correctly as S. algae by 16S rRNA gene sequence analysis.


Subject(s)
Gram-Negative Bacterial Infections/microbiology , Osteomyelitis/microbiology , Shewanella/classification , Shewanella/isolation & purification , Bacterial Typing Techniques , DNA, Bacterial/analysis , DNA, Ribosomal/analysis , Female , Genes, rRNA , Humans , Middle Aged , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Shewanella/genetics
16.
Science ; 293(5537): 2093-8, 2001 Sep 14.
Article in English | MEDLINE | ID: mdl-11557893

ABSTRACT

Rickettsia conorii is an obligate intracellular bacterium that causes Mediterranean spotted fever in humans. We determined the 1,268,755-nucleotide complete genome sequence of R. conorii, containing 1374 open reading frames. This genome exhibits 804 of the 834 genes of the previously determined R. prowazekii genome plus 552 supplementary open reading frames and a 10-fold increase in the number of repetitive elements. Despite these differences, the two genomes exhibit a nearly perfect colinearity that allowed the clear identification of different stages of gene alterations with gene remnants and 37 genes split in 105 fragments, of which 59 are transcribed. A 38-kilobase sequence inversion was dated shortly after the divergence of the genus.


Subject(s)
Evolution, Molecular , Genome, Bacterial , Rickettsia conorii/genetics , Rickettsia prowazekii/genetics , Adaptation, Physiological , Chlamydia/genetics , Computational Biology , DNA, Bacterial/genetics , DNA, Intergenic , Gene Dosage , Gene Silencing , Gene Transfer, Horizontal , Genes, Bacterial , Open Reading Frames , Phylogeny , Polymerase Chain Reaction , Repetitive Sequences, Nucleic Acid , Rickettsia/genetics , Rickettsia conorii/physiology , Rickettsia prowazekii/physiology , Sequence Analysis, DNA , Transcription, Genetic
17.
Int J Syst Evol Microbiol ; 51(Pt 4): 1353-1360, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11491333

ABSTRACT

'Gene D' is the PS120-protein-encoding gene, first described in Rickettsia conorii and Rickettsia japonica. Sequence analysis of a 3030 bp fragment of 'gene D' in 24 representatives of the genus Rickettsia was carried out to complete phylogenetic analyses previously inferred by comparison of gene sequences encoding citrate synthase, 17 kDa antigen and rOmpA and rOmpB. The phylogenetic relationships between rickettsiae were inferred from the comparison of both the gene and the derived protein sequences, using the parsimony, neighbour-joining and maximum-likelihood methods. Five distinct groups of rickettsiae were identified. These were: the Rickettsia massiliae group, including R. massiliae, Bar 29, Rickettsia rhipicephali and Rickettsia aeschlimannii; the Rickettsia rickettsii group containing Rickettsia sibirica, 'Rickettsia mongolotimonae', Rickettsia parkeri, strain S, Rickettsia africae, the R. conorii complex, Rickettsia slovaca, Rickettsia honei, R. rickettsii, R. japonica and Rickettsia montanensis; the group currently containing only Rickettsia helvetica; the Rickettsia akari group including Rickettsia australis, R. akari and the ELB agent; Rickettsia prowazekii and Rickettsia typhi clustered in the typhus group. As significant bootstrap values were obtained for most of the nodes, sequence comparison of 'gene D' should be considered as a complementary approach in phylogenetic studies of rickettsiae.


Subject(s)
Bacterial Proteins/genetics , Genes, Bacterial , Rickettsia/classification , Rickettsia/genetics , Molecular Sequence Data , Phylogeny , Species Specificity
18.
Emerg Infect Dis ; 7(1): 73-81, 2001.
Article in English | MEDLINE | ID: mdl-11266297

ABSTRACT

A rickettsia named the ELB agent, or "Rickettsia felis," was identified by molecular biology techniques in American fleas in 1990 and later in four patients from Texas and Mexico. We attempted to isolate this rickettsia from infected fleas at various temperatures and conditions. A representative isolate of the ELB agent, the Marseille strain, was characterized and used to develop a microimmunofluorescence test that detected reactive antibodies in human sera. The ELB agent was isolated from 19 of 20 groups of polymerase chain reaction-proven infected fleas. The microimmunofluorescence results provided serologic evidence of infection by the ELB agent in four patients with fever and rash in France (2) and Brazil (2), supporting the pathogenic role of this rickettsia. Our successful isolation of this rickettsia makes it available for use in serologic tests to determine its clinical spectrum, prevalence, and distribution.


Subject(s)
Rickettsia/isolation & purification , Siphonaptera/microbiology , Animals , Humans , Phylogeny , Polymerase Chain Reaction , Rickettsia/classification
19.
J Clin Microbiol ; 39(2): 430-7, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11158086

ABSTRACT

Identification of Bartonella species is of increasing importance as the number of infections in which these bacteria are involved increases. To date, these gram-negative bacilli have been identified by various serological, biochemical, and genotypic methods. However, the development of alternative tools is required, principally to circumvent a major risk of contamination during sample manipulation. The aim of our study was to investigate the possible identification of various Bartonella species by comparison of RNA polymerase beta-subunit gene (rpoB) sequences. This approach has previously been shown to be useful for the identification of members of the family Enterobacteriaceae (C. M. Mollet, M. Drancourt, and D. Raoult, Mol. Microbiol. 26:1005-1011, 1997). Following PCR amplification with specific oligonucleotides, a 825-bp region of the rpoB gene was sequenced from 13 distinct Bartonella strains. Analysis of these sequences allowed selection of three restriction enzymes (ApoI, AluI, and AflIII) useful for discerning the different strains by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis. To confirm the potential value of such an approach for identification of Bartonella, the rpoB PCR was then applied to 94 clinical samples, and the results obtained were identical to those obtained by our reference PCR method. Twenty-four isolates were also adequately identified by PCR-RFLP analysis. In all cases, our results were in accordance with those of the reference method. Moreover, conserved regions of DNA were chosen as suitable primer targets for PCR amplification of a 439-bp fragment which can be easily sequenced.


Subject(s)
Bartonella Infections/diagnosis , Bartonella/classification , DNA-Directed RNA Polymerases/genetics , Animals , Bartonella/genetics , Bartonella/isolation & purification , Bartonella Infections/microbiology , Bartonella henselae/classification , Bartonella henselae/isolation & purification , Bartonella quintana/classification , Bartonella quintana/isolation & purification , Base Sequence , Cats , DNA, Bacterial/blood , DNA, Bacterial/genetics , Humans , Lymph Nodes/microbiology , Molecular Sequence Data , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Sequence Alignment , Sequence Homology, Nucleic Acid , Suppuration/microbiology
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