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1.
Emerg Infect Dis ; 29(2)2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36564155

RESUMO

We report intrafamilial transmission of monkeypox virus to all members of a family (father, mother, and 2 children). Case reports in young children have been extremely rare during the 2022 mpox outbreak. Their clinical signs were mild, and clinical diagnosis would be difficult without knowledge of the father's monkeypox virus infection.


Assuntos
Monkeypox virus , Mpox , Humanos , Criança , Pré-Escolar , Monkeypox virus/genética , Mpox/diagnóstico , Mpox/epidemiologia , Surtos de Doenças
2.
Acta Derm Venereol ; 99(2): 170-174, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30328468

RESUMO

Bacteraemias are life-threatening conditions that require rapid care and antibiotherapy. Dermatological signs might help in deciding the most relevant treatment. The aim of this study was to determine the prevalence and clinical characteristics of cutaneous manifestations in hospitalized patients with bacteraemia. A cross-sectional study was conducted over a period of 1 year. All consecutive patients with a bacteraemia (except contaminations) were included and examined by a dermatologist within 48 h after positive blood cultures. Clinical (skin manifestations, diagnosis, origin of the bacteraemia) and laboratory (bacteria) data were recorded. In total, 401 bacteraemias in 375 patients were included for the final analysis. Thirty-nine cutaneous manifestations in 34 patients were noted, corresponding to a prevalence of 9%; 69% (n = 27) were considered primary cutaneous manifestations, 18% (n = 7) as secondary ones, 10% (n = 4) as contiguous, and 3% (n = 1) as undetermined. Gram-positive cocci, specifically Staphylococcus aureus and Streptococcus species, were the most frequent bacteria (n = 27, 69%).


Assuntos
Bacteriemia/epidemiologia , Bacteriemia/microbiologia , Dermatopatias Bacterianas/epidemiologia , Dermatopatias Bacterianas/microbiologia , Pele/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Bacteriemia/diagnóstico , Criança , Estudos Transversais , Feminino , França/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Prospectivos , Dermatopatias Bacterianas/diagnóstico , Infecções Cutâneas Estafilocócicas/epidemiologia , Infecções Cutâneas Estafilocócicas/microbiologia , Infecções Estreptocócicas/epidemiologia , Infecções Estreptocócicas/microbiologia , Fatores de Tempo , Adulto Jovem
3.
BMC Microbiol ; 14: 325, 2014 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-25551282

RESUMO

BACKGROUND: The very-preterm infant gut microbiota is increasingly explored due to its probable role in the development of life threatening diseases. Results of high-throughput studies validate and renew the interest in approaches with lower resolution such as PCR-Temporal Temperature Gel Electrophoresis (TTGE) for the follow-up of dominant microbiota dynamics. We report here an extensive longitudinal study of gut colonization in very preterm infants. We explored by 16S rDNA-based PCR-TTGE a total of 354 stool specimens sampled during routine monitoring from the 1(st) to the 8(th) week of life in 30 very pre-term infants born before 30 weeks of gestational age. RESULTS: Combining comparison with a diversity ladder and sequencing allowed affiliation of 50 Species-Level Operational Taxonomic Units (SLOTUs) as well as semi-quantitative estimation of Operational Taxonomic Units (OTUs). Coagulase-negative staphylococci, mainly the Staphylococcus epidermidis, was found in all the infants during the study period and was the most represented (75.7% of the SLOTUs) from the first days of life. Enterococci, present in 60% of the infants were early, highly represented and persistent colonizers of the premature gut. Later Enterobacteriaceae and the genus Clostridium appeared and were found in 10 (33%) and 21 infants (70%), respectively. We showed a high representation of Veillonella in more than a quarter of the infants and being able to persistently colonize premature gut. The genera Anaerococcus, Aquabacterium, Bacillus, Bifidobacterium, Corynebacterium, Micrococcus, Oceanobacillus, Propionibacterium, Pseudomonas, Rothia, Sarcina, Sneathia and Streptococcus were observed as transient or persistent colonizers, each genus being found in a minority of infants. CONCLUSIONS: Despite low resolution, PCR-TTGE remains complementary to high-throughput sequencing-based approaches because it allows the follow-up of dominant bacteria in gut microbiota in a large longitudinal cohorts of preterm neonates. We described the development of pre-term gut microbiota that should be now replaced regarding the functional role of major OTUs.


Assuntos
Microbioma Gastrointestinal , Trato Gastrointestinal/microbiologia , Recém-Nascido Prematuro , Microbiota , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Fezes/microbiologia , Humanos , Lactente , Recém-Nascido , Estudos Longitudinais , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Fatores de Tempo
4.
Dermatology ; 222(2): 167-70, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21346325

RESUMO

BACKGROUND/AIMS: Panton-Valentine leukocidin (PVL)-positive Staphylococcus aureus have been associated with suppurative infections; however, their precise role in skin infections has not been elucidated. We studied the rate of PVL-positive S. aureus in the different types of skin infections and compared follicular to nonfollicular skin infections. METHODS: In our institution, patients with a skin infection caused by S. aureus were enrolled in a prospective, observational cohort study (from July 1, 2003 to June 30, 2010). We studied the rate of PVL-positive S. aureus in the different clinical types of skin infections and compared the rate of PVL-positive S. aureus in follicular infections to that in nonfollicular infections. RESULTS: A total 229 skin infections were included: 97 (42.5%) were caused by PVL-positive strains. Thirty-nine of the 53 (74%) follicular infections [8 of the 17 (47%) with folliculitis, 30 of the 35 (85.5%) with furuncles and 1 with a carbuncle (100%)] were caused by PVL-positive S. aureus, compared to 16 of the 131 (12%) nonfollicular infections (p < 0.001). CONCLUSION: PVL-positive S. aureus strains are mainly associated with follicular skin infections.


Assuntos
Toxinas Bacterianas/análise , Exotoxinas/análise , Doenças do Cabelo/microbiologia , Folículo Piloso/microbiologia , Leucocidinas/análise , Infecções Cutâneas Estafilocócicas/microbiologia , Staphylococcus aureus/isolamento & purificação , Humanos , Estudos Prospectivos
5.
Microbiology (Reading) ; 156(Pt 7): 2080-2091, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20413553

RESUMO

As well as intraspecific heterogeneity, intragenomic heterogeneity between 16S rRNA gene copies has been described for a range of bacteria. Due to the wide use of 16S rRNA gene sequence analysis for taxonomy, identification and metagenomics, evaluating the extent of these heterogeneities in natural populations is an essential prerequisite. We investigated inter- and intragenomic 16S rRNA gene heterogeneity of the variable region V3 in a population of 149 clinical isolates of Veillonella spp. of human origin and in 13 type or reference Veillonella strains using PCR-temporal temperature gel electrophoresis (TTGE). 16S rRNA gene diversity was high in the studied population, as 45 different banding patterns were observed. Intragenomic heterogeneity was demonstrated for 110 (74 %) isolates and 8 (61.5 %) type or reference strains displaying two or three different gene copies. Polymorphic nucleotide positions accounted for 0.5-2.5 % of the sequence and were scattered in helices H16 and H17 of the rRNA molecule. Some of them changed the secondary structure of H17. Phylotaxonomic structure of the population based on the single-copy housekeeping gene rpoB was compared with TTGE patterns. The intragenomic V3 heterogeneity, as well as recombination events between strains or isolates of different rpoB clades, impaired the 16S rRNA-based identification for some Veillonella species. Such approaches should be conducted in other bacterial populations to optimize the interpretation of 16S rRNA gene sequences in taxonomy and/or diversity studies.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Variação Genética , RNA Ribossômico 16S/genética , Veillonella/classificação , Veillonella/isolamento & purificação , Proteínas de Bactérias/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Dados de Sequência Molecular , Veillonella/genética
6.
Dermatology ; 219(4): 299-302, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19648730

RESUMO

BACKGROUND: The role of Panton-Valentine leukocidin (PVL) in skin and soft-tissue infections is not clear. OBJECTIVE: Our purpose was to determine the prevalence of PVL gene carriage among Staphylococcus aureus strains isolated from primary and secondary skin abscesses. METHODS: A prospective study was conducted. From July 2003 to June 2008, S. aureus isolates from skin abscesses were screened for the PVL genes. The abscesses were considered primary if they occurred on previously healthy skin and secondary in all other cases. RESULTS: Fifty-seven patients presenting with S. aureus skin abscesses were included in the study. The PVL genes were detected in 40 (70%) of the 57 S. aureus isolates. Thirty-eight (92.7%) of the 41 primary skin abscesses were due to PVL-positive strains, compared to only 2 (12.5%) of the 16 secondary skin abscesses (p < 0.001). CONCLUSIONS: Primary skin abscesses are mainly caused by PVL-positive S. aureus strains.


Assuntos
Abscesso/microbiologia , Abscesso/patologia , Toxinas Bacterianas/genética , Exotoxinas/genética , Leucocidinas/genética , Infecções Cutâneas Estafilocócicas/microbiologia , Infecções Cutâneas Estafilocócicas/patologia , Staphylococcus aureus/genética , Abscesso/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , França/epidemiologia , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Prospectivos , Infecções Cutâneas Estafilocócicas/epidemiologia , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/patogenicidade
7.
Pediatr Infect Dis J ; 38(9): e230-e232, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31261364

RESUMO

We described nosocomially clustered cases of measles secondary to a nonvaccinated index case occurring in a teenage psychiatric unit despite optimum vaccine coverage. Surveillance of this fully vaccinated closed cohort showed a 7% attack rate. Vaccination limited the risk of complicated measles and the onset of a large outbreak.


Assuntos
Infecção Hospitalar/transmissão , Infecção Hospitalar/virologia , Surtos de Doenças/prevenção & controle , Vacina contra Sarampo/administração & dosagem , Sarampo/transmissão , Vacinação/estatística & dados numéricos , Adolescente , Criança , Estudos de Coortes , Monitoramento Epidemiológico , Feminino , França/epidemiologia , Hospitais , Humanos , Incidência , Sarampo/epidemiologia , Sarampo/prevenção & controle , Unidade Hospitalar de Psiquiatria/estatística & dados numéricos
8.
Int J Antimicrob Agents ; 53(5): 678-681, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30742957

RESUMO

BACKGROUND: Increasing incidence of hepatitis C virus (HCV) infection in human immunodeficiency virus (HIV)-positive men having sex with men (MSM) has been described in recent years. Phylogenetic analyses of acute HCV infections were undertaken to characterize the dynamics during the epidemic in Paris, and associated sexually transmitted infections (STIs) were evaluated. METHODS: Sanger sequencing of polymerase gene was performed. Maximum likelihood phylogenies were reconstructed using FastTree 2.1 under a GTR+CAT model. Transmission chains were defined as clades with a branch probability ≥0.80 and intraclade genetic distances <0.02 nucleotide substitutions per sites. STIs detected ≤1 month before HCV diagnosis were considered. RESULTS: Among the 85 studied patients, at least 81.2% were MSM. Respectively, 47.6%, 39.0%, 11.0% and 2.4% were infected with genotypes 1a, 4d, 3a and 2k. At least 91.8% were co-infected with HIV. HCV re-infection was evidenced for 24.7% of patients and STIs for 20.0% of patients. Twenty-two transmission chains were identified, including 52 acute hepatitis C (11 pairs and 11 clusters from three to seven patients). CONCLUSIONS: These results revealed strong clustering of acute HCV infections. Thus, rapid treatment of both chronic and acute infections is needed among this population to decrease the prevalence of HCV, in combination with preventive behavioural interventions.


Assuntos
Análise por Conglomerados , Transmissão de Doença Infecciosa , Infecções por HIV/complicações , Hepacivirus/isolamento & purificação , Hepatite C/epidemiologia , Infecções Sexualmente Transmissíveis/epidemiologia , Adulto , Genótipo , Hepacivirus/classificação , Hepacivirus/genética , Hepatite C/transmissão , Homossexualidade Masculina , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Paris/epidemiologia , Filogenia , Prevalência , Estudos Retrospectivos , Análise de Sequência de DNA , Infecções Sexualmente Transmissíveis/transmissão
10.
AIDS ; 20(12): 1627-35, 2006 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-16868444

RESUMO

BACKGROUND: Osteonecrosis was increasingly associated with HIV infection in the 1990s. It is unclear whether its risk increases with the duration of HIV infection, the duration of combination antiretroviral therapy (cART) or both. OBJECTIVE: To analyse factors associated with the rate of symptomatic osteonecrosis, particularly the relative impacts of the duration of HIV infection and the duration of cART, using the French Hospital Database on HIV, which comprises a large number of subjects with substantial follow-up. METHODS: Poisson regression model was used to identify factors associated with the rate of osteonecrosis among patients enrolled in 1996-2002. RESULTS: The study involved 56,393 subjects with a total follow-up of 229,031 person-years. Symptomatic osteonecrosis was diagnosed in 104 subjects with an incidence rate of 4.5/10,000 person-years. Multivariate analysis identified three factors associated with the rate of osteonecrosis: prior AIDS-defining illnesses [adjusted relative rate (RR), 3.1; 95% confidence interval (CI), 2.0-4.9], the CD4 cell nadir [RR, 1.6 (95% CI, 0.9-2.9) for CD4 cell count 50-199 cells/microl; RR, 1.8 (95% CI, 1.0-3.3) for CD4 cell count < 50 cells/microl; both relative to CD4 cell count > or = 200 cells/microl] and exposure to cART. Compared with unexposed patients, the RR of osteonecrosis ranged from 2.6 (95% CI, 1.2-5.9) in patients treated with cART for < 12 months to 5.1 (95% CI, 2.1-12.6) in patients treated for > or = 60 months. CONCLUSIONS: Osteonecrosis appears to be a complication of both HIV infection and cART.


Assuntos
Antirretrovirais/efeitos adversos , Infecções por HIV/tratamento farmacológico , Osteonecrose/etiologia , Adulto , Contagem de Linfócito CD4 , Bases de Dados Factuais , Quimioterapia Combinada , Feminino , França/epidemiologia , Infecções por HIV/complicações , Infecções por HIV/epidemiologia , Humanos , Incidência , Masculino , Modelos Estatísticos , Osteonecrose/epidemiologia , Fatores de Risco , Fatores de Tempo
11.
Clin Infect Dis ; 40(2): 306-8, 2005 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-15655752

RESUMO

Cryoglobulinemia was studied in human immunodeficiency virus-positive, hepatitis C virus (HCV)-positive patients in relation to their CD4 cell count. Cryoglobulinemia was found in 18 (31.6%) of 57 patients: 17 (44.7%) of 38 patients with a CD4 cell count of >or=200 cells/ micro L versus 1 (5.3%) of 19 patients with a CD4 cell count of <200 cells/ micro L (P=.0064). Cell-mediated immunity could, therefore, contribute to the production of HCV-associated cryoglobulins.


Assuntos
Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos/citologia , Crioglobulinemia/complicações , Infecções por HIV/complicações , Hepatite C/complicações , Adulto , Linfócitos T CD4-Positivos/imunologia , Crioglobulinemia/sangue , Crioglobulinemia/etiologia , Feminino , Infecções por HIV/sangue , Hepatite C/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Carga Viral
12.
J Clin Virol ; 32(3): 241-4, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15722030

RESUMO

The human immunodeficiency virus type 1 (HIV-1) resistance profile, K65R, K70E and M184V, on reverse transcriptase gene was associated with the virologic rebound consecutively to the switch of lopinavir/r to tenofovir DF in a stable regimen with nucleoside backbone of abacavir, lamivudine and didanosine. The high selective pressure on the same resistance pathway was probably associated with the loss of antiviral potency, even in well-controlled patient.


Assuntos
Adenina/análogos & derivados , Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/virologia , HIV-1/efeitos dos fármacos , Organofosfonatos/uso terapêutico , Adenina/uso terapêutico , Adulto , Didanosina/uso terapêutico , Didesoxinucleosídeos/uso terapêutico , Farmacorresistência Viral/genética , Feminino , Gabão , Infecções por HIV/tratamento farmacológico , Transcriptase Reversa do HIV/genética , HIV-1/genética , Humanos , Lamivudina/uso terapêutico , Lopinavir , Testes de Sensibilidade Microbiana , Mutação , Pirimidinonas/uso terapêutico , Tenofovir , Carga Viral
13.
Clin Infect Dis ; 35(7): 880-2, 2002 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-12228826

RESUMO

We report the case histories of 7 human immunodeficiency virus (HIV)-infected patients receiving highly active antiretroviral therapy (HAART) who had a diagnosis of Castleman disease. All 6 patients who were treated responded to chemotherapy; immune reconstitution was observed in 5 patients, but it did not prevent relapse of Castleman disease. However, the mean duration of survival observed in this series (48 months) was most probably due to immune reconstitution resulting from receipt of HAART, which reduced the mortality associated with HIV disease.


Assuntos
Hiperplasia do Linfonodo Gigante/etiologia , Infecções por HIV/tratamento farmacológico , Infecções por Herpesviridae/etiologia , Herpesvirus Humano 8 , Adulto , Terapia Antirretroviral de Alta Atividade , Hiperplasia do Linfonodo Gigante/mortalidade , Infecções por HIV/complicações , Infecções por HIV/imunologia , Infecções por HIV/mortalidade , Infecções por Herpesviridae/mortalidade , Herpesvirus Humano 8/imunologia , Herpesvirus Humano 8/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva , Análise de Sobrevida
17.
J Infect ; 64(6): 613-7, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22230736

RESUMO

We report the first case of Mycobacterium avium reactivation, after prolonged latency, in a HIV-infected patient receiving highly active antiretroviral therapy with undetectable viral replication and normal CD4 cell count. The patient presented with a painful swollen shoulder seven years after initial M. avium bacteriaemia. Articular puncture grew M. avium. The isolates of the first and second infection were identical using repetitive-sequence-based Polymerase Chain Reaction analyses.


Assuntos
Terapia Antirretroviral de Alta Atividade , Infecções por HIV/complicações , Infecções por HIV/tratamento farmacológico , Mycobacterium avium/isolamento & purificação , Tuberculose/diagnóstico , Tuberculose/microbiologia , Contagem de Linfócito CD4 , DNA Bacteriano/genética , Genótipo , HIV/isolamento & purificação , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Tipagem Molecular , Infecções por Mycobacterium , Reação em Cadeia da Polimerase , Radiografia , Recidiva , Ombro/diagnóstico por imagem , Ombro/patologia , Tuberculose/patologia , Carga Viral
18.
Res Microbiol ; 161(2): 91-100, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20079831

RESUMO

Members of the phylum Synergistetes have been demonstrated in several environmental ecosystems and mammalian microflorae by culture-independent methods. In the past few years, the clinical relevance of some uncultivated phylotypes has been demonstrated in endodontic infections, and uncultured Synergistetes have been demonstrated in human mouth, gut and skin microbiota. However, Synergistetes are rarely cultured from human samples, and only 17 isolates are currently reported. Twelve members of Synergistetes isolated in the course of various infectious processes, including 3 Jonquetella anthropi, 2 Cloacibacillus evryensis, 2 Pyramidobacter piscolens and 5 unidentified strains, as well as 56 clones obtained by specific PCR from the normal vaginal microflora, were studied. 16S rRNA gene-based phylogeny showed that the clones were grouped into 3 clusters, corresponding to the genus Jonquetella, P. piscolens and one novel Synergistetes taxon. The presence and diversity of Synergistetes were reported for the first time in the vaginal microflora. Synergistetes were found in healthy patients, suggesting that they could play a functional role in human microflorae, but may also act as opportunistic pathogens. Studying the phylogenetic relationships between environmental and mammalian strains and clones revealed clearly delineated independent lineages according to the origin of the sequences.


Assuntos
Bactérias/classificação , Bactérias/genética , Infecções Bacterianas/microbiologia , Portador Sadio/microbiologia , Filogenia , Polimorfismo Genético , Bactérias/isolamento & purificação , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Feminino , Humanos , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vagina/microbiologia
19.
Int J Syst Evol Microbiol ; 59(Pt 5): 1028-35, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19406787

RESUMO

The number of bacterial phyla has greatly increased in the past decade. Among them, a candidate division named 'Synergistes' was proposed in a phylogenetic study on the global diversity of bacteria. We previously described the genus Jonquetella and suggested that it belonged to this not yet well-delineated candidate phylum. 16S rRNA gene based-phylogeny studies were conducted using four reconstruction methods and 599 sequences forming five datasets were used in an alternative treeing approach. These analyses indicated that the genera Aminiphilus, Aminobacterium, Aminomonas, Anaerobaculum, Dethiosulfovibrio, Jonquetella, Synergistes, Thermanaerovibrio and Thermovirga should be grouped in the same high-level taxon. This taxon was shown to be a phylum-rank lineage in the domain Bacteria and, because of the prior use of the name Synergistes for a genus, the name 'Synergistetes' is proposed for this candidate phylum. We also propose an emended delineation of the phylum 'Deferribacteres', which is now only represented by the family Deferribacteriaceae. The emended family Syntrophomonadaceae is limited to the genera Pelospora, Syntrophomonas, Syntrophothermus and Thermosyntropha.


Assuntos
Bactérias/classificação , Bactérias/genética , Filogenia , Terminologia como Assunto , Genes de RNAr , Variação Genética , RNA Ribossômico 16S/genética
20.
J Microbiol Methods ; 79(2): 156-65, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19686785

RESUMO

Temporal Temperature Gel Electrophoresis of amplified 16S rRNA gene sequences (16S rDNA PCR-TTGE) constitutes a culture-independent molecular method used to study bacterial communities. All the technical steps are crucial for quality and exhaustiveness of the results obtained by such approach. Careful optimization of the protocols used is ideally needed for each ecosystem studied. We present here the strategy used to construct an optimized protocol for a 16S rDNA PCR-TTGE-based analysis of gut microflora in neonates. Improvement of the different steps, i.e. total DNA extraction, amplification in terms of efficiency and reduction of heteroduplex formation, TTGE migration conditions and bacterial identification from TTGE patterns, was performed. The optimized protocol was used for the subsequent analysis of 14 stool samples comparatively to a culture-based method. We showed that a specifically designed ladder representative of the diversity of the studied microflora is a useful tool for the identification of bacterial taxa despite biases inherent to 16S rRNA genes, including intra-genomic heterogeneity. Cultivation and PCR-TTGE gave congruent results but cultivation was more efficient for the detection of minor populations whereas PCR-TTGE gave a more complete description of the major populations. Finally, we demonstrated the reliability, the detection sensitivity and the convenience of the optimized 16S rDNA PCR-TTGE method compared with cultural approaches for studying the premature neonate gut microbiota.


Assuntos
Bactérias/classificação , Bactérias/isolamento & purificação , Técnicas Bacteriológicas/métodos , Biodiversidade , Eletroforese em Gel de Poliacrilamida/métodos , Trato Gastrointestinal/microbiologia , Reação em Cadeia da Polimerase/métodos , Bactérias/genética , Bactérias/crescimento & desenvolvimento , DNA Bacteriano/genética , DNA Ribossômico/genética , Fezes/microbiologia , Feminino , Temperatura Alta , Humanos , Recém-Nascido , Masculino , Desnaturação de Ácido Nucleico , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade
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