RESUMO
BACKGROUND: Dengue (DENV) and Zika virus (ZIKV) are important mosquito-transmitted viruses. OBJECTIVES: To investigate the performance of Standard F, Fluorescence Immunoassay (FIA, SD Biosensor Inc., Suwon, South Korea) providing results in 15 min to detect DENV IgG, IgM and NS1Ag, and ZIKV IgG, IgM, and Ag. STUDY DESIGN: A well-characterized panel of patient samples (11 acute DENV, 11 acute ZIKV, 10 past DENV, 10 past ZIKV infection, 36 with other conditions) were tested with the FIA test. RESULTS: In acute DENV infection, the combination of FIA-NS1Ag and/or IgM positivity showed a sensitivity of 100%. In past DENV, FIA-IgG test showed a sensitivity of 70%. Specificity of FIA-DENV NS1Ag, IgG, and IgM was 87.5%, 83.5%, and 91.7%, respectively. The sensitivity of FIA-ZIKV IgM and FIA-ZIKV Ag, in confirmed acute infection, was 72.7% and 9.1%, respectively. FIA-ZIKV Ag did not improve the sensitivity in detecting acute ZIKV infection, being positive only in one IgM positive sample. In past ZIKV infection (32-183 days after symptom onset), FIA-ZIKV IgG and IgM showed a sensitivity of 40% and 80% respectively, generating an overall 90% sensitivity. Specificity of FIA-ZIKV Ag, IgM, and IgG was 92.6%, 100%, and 97%, respectively. CONCLUSION: FIA test, a rapid and easy to perform assay, showed high sensitivity to detect acute DENV infection, but lower in acute ZIKV infection. In past ZIKV infections, the best performance of FIA test is obtained by combining detection of IgG and IgM.
Assuntos
Anticorpos Antivirais/sangue , Dengue/diagnóstico , Imunofluorescência , Infecção por Zika virus/diagnóstico , Antígenos Virais/imunologia , Reações Cruzadas , Dengue/imunologia , Vírus da Dengue , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Sensibilidade e Especificidade , Testes Sorológicos , Zika virus , Infecção por Zika virus/imunologiaRESUMO
The characterization of 2 clinical isolates of Mycobacterium sherrisii provides further phenotypic and genotypic information beyond that reported in the article that originally described this species. One of our strains was responsible for pulmonary disease in a middle-aged non-HIV patient; thus, confirming the potential pathogenicity of this species previously reported only in an HIV-positive patient.
Assuntos
Pneumopatias/diagnóstico , Pneumopatias/microbiologia , Mycobacterium/classificação , Mycobacterium/isolamento & purificação , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Chaperonina 60 , Chaperoninas/genética , DNA Bacteriano/análise , DNA Espaçador Ribossômico/análise , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mycobacterium/genética , Infecções por Mycobacterium/diagnóstico , Infecções por Mycobacterium/microbiologia , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , Análise de Sequência de DNARESUMO
A non-tuberculous mycobacterium was isolated, following a vertebral needle aspiration, from the blood of a patient with severe spondylodiscitis. The strain turned out to be different from any known mycobacterial species and was quite drug-susceptible in vitro. The patient improved markedly following treatment with meropenem, clarithromycin and amikacin.
Assuntos
Antituberculosos/uso terapêutico , Discite/tratamento farmacológico , Discite/microbiologia , Mycobacterium/efeitos dos fármacos , Mycobacterium/isolamento & purificação , Sequência de Bases , DNA Espaçador Ribossômico/análise , Genes de RNAr , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mycobacterium/classificação , Mycobacterium/genética , Infecções por Mycobacterium/tratamento farmacológico , Infecções por Mycobacterium/microbiologia , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , Análise de Sequência de DNA , Resultado do TratamentoAssuntos
Pessoal de Laboratório Médico/psicologia , Segurança do Paciente/legislação & jurisprudência , Gestão de Riscos/ética , Carga de Trabalho/psicologia , Local de Trabalho/normas , Patógenos Transmitidos pelo Sangue/isolamento & purificação , Infecção Hospitalar/prevenção & controle , Testes Diagnósticos de Rotina/ética , Testes Diagnósticos de Rotina/estatística & dados numéricos , Hepatite B/sangue , Hepatite B/diagnóstico , Antígenos da Hepatite B/sangue , Antígenos da Hepatite B/genética , Antígenos da Hepatite B/imunologia , Humanos , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Itália/epidemiologia , Pessoal de Laboratório Médico/economia , Pessoal de Laboratório Médico/normas , Isolamento de Pacientes/normas , Segurança do Paciente/normas , Diálise Renal/métodos , Local de Trabalho/estatística & dados numéricosRESUMO
Four bacterial strains were isolated from independent clinical specimens in different countries and their genotypic and phenotypic characters support their classification in a novel species within the genus Mycobacterium. One strain was clearly responsible for a severe, post-traumatic wound infection in a healthy boy. The novel species, for which the name Mycobacterium monacense sp. nov. is proposed, is yellow-pigmented, non-photochromogenic and grows in less than a week on solid medium. Based on phenotypic investigations alone, distinction of these four strains from known scotochromogenic rapidly growing strains is problematic. However, the novel strains differ from any other mycobacterium in each of the molecular species markers investigated: the 16S rRNA gene, the 16S-23S rRNA gene internal transcribed spacer and the hsp65 gene. Of the strains investigated, two different sequevars were detected for the hsp65 region. Phylogenetic analysis revealed that these four strains were most closely related to Mycobacterium doricum. The type strain of Mycobacterium monacense sp. nov. is B9-21-178T (=DSM 44395T=CIP 109237T).
Assuntos
Infecções por Mycobacterium não Tuberculosas/microbiologia , Ácidos Micólicos/análise , Micobactérias não Tuberculosas/classificação , Micobactérias não Tuberculosas/isolamento & purificação , Adulto , Idoso de 80 Anos ou mais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Líquido da Lavagem Broncoalveolar/microbiologia , Chaperonina 60 , Chaperoninas/genética , Criança , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Feminino , Genes de RNAr , Alemanha , Humanos , Itália , Masculino , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Micobactérias não Tuberculosas/fisiologia , Filogenia , Pigmentos Biológicos , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Escarro/microbiologia , Infecção dos Ferimentos/microbiologiaRESUMO
We report on four cases of infection by the recently described species Mycobacterium parascrofulaceum. In two cases the mycobacterium was isolated from AIDS patients, while in the others it was responsible for pulmonary disease in elderly men. Our findings suggest that M. parascrofulaceum is an opportunistic pathogen, like many other nontuberculous mycobacterial species.
Assuntos
Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Infecções por Mycobacterium/microbiologia , Mycobacterium/isolamento & purificação , Infecções Oportunistas/microbiologia , Adulto , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Mycobacterium/classificação , Mycobacterium/genéticaRESUMO
Eight mycobacterial strains isolated during an 11 year period from the sputum of independent patients with various pulmonary disorders and, in one case, from a lymph node of a young girl, were found to present identical features. Phenotypic and genotypic characteristics revealed that the most closely related species to these test isolates were Mycobacterium triplex and Mycobacterium lentiflavum. However, the lipids of the cell wall of the test isolates differed from those of the latter species by TLC and presented unique profiles by both GC and HPLC. Genotypic analysis showed that they had unique 16S rRNA gene and internal transcribed spacer (ITS) sequences, and could be differentiated from all other mycobacterial strains by PCR restriction analysis of hsp65. The strains presented high resistance to antimycobacterial drugs. The name Mycobacterium florentinum sp. nov. is proposed for this taxon, with strain FI-93171(T) (=DSM 44852(T) = CIP 108409(T)) as the type strain.
Assuntos
Linfonodos/microbiologia , Mycobacterium/classificação , Mycobacterium/isolamento & purificação , Escarro/microbiologia , Adulto , Idoso , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Parede Celular/química , Chaperonina 60 , Chaperoninas/genética , Criança , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico , Farmacorresistência Bacteriana , Fezes/microbiologia , Feminino , Genes de RNAr , Humanos , Lipídeos/análise , Lipídeos/isolamento & purificação , Masculino , Dados de Sequência Molecular , Mycobacterium/citologia , Mycobacterium/fisiologia , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
INNO-LiPA MYCOBACTERIA (Innogenetics, Ghent, Belgium) is a reverse hybridization DNA probe assay that has been recently improved by increasing the number of identifiable mycobacterial species to 16. Our assessment, performed with 197 mycobacteria belonging to 81 taxa, revealed 100% specificity and sensitivity for 20 out of 23 probes. The probes specific for Mycobacterium fortuitum complex, for the Mycobacterium avium-intracellulare-scrofulaceum group, and for Mycobacterium intracellulare type 2 cross-reacted with several mycobacteria rarely isolated from clinical specimens. The overall sensitivity was 100%, and the overall specificity was 94.4%.
Assuntos
Sondas de DNA , Mycobacterium/isolamento & purificação , Hibridização de Ácido Nucleico , Mycobacterium/classificação , Sensibilidade e EspecificidadeRESUMO
Mycobacterium triplex, a recently described, potentially pathogenic species, caused disease primarily in immunocompromised patients. We report a case of pulmonary infection due to this mycobacterium in an immunocompetent patient and review the characteristics of two other cases. In our experience, Mycobacterium triplex pulmonary infection is unresponsive to antimycobacterial chemotherapy.
Assuntos
Pneumopatias/microbiologia , Infecções por Mycobacterium/microbiologia , Mycobacterium , Idoso , Antibacterianos/uso terapêutico , Sequência de Bases , DNA Bacteriano/análise , Feminino , Humanos , Imunocompetência , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mycobacterium/classificação , Infecções por Mycobacterium/tratamento farmacológico , Infecções por Mycobacterium/imunologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
A lymph node excision was performed on a 45-year-old woman with left cervical swelling. The disorder which developed after the patient had undergone oral surgery for a severe periodontal disease failed to respond to antimicrobial chemotherapy. A mycobacterial strain subsequently identified by high-performance liquid chromatography analysis of cell wall mycolic acids as Mycobacterium lentiflavum grew from the excised specimen. This case and previously published reports highlight the relevance of M. lentiflavum as an emerging causative agent of mycobacterial cervical lymphadenitis.
Assuntos
Mycobacterium/isolamento & purificação , Tuberculose dos Linfonodos/diagnóstico , Sequência de Bases , Parede Celular/química , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Excisão de Linfonodo , Pessoa de Meia-Idade , Mycobacterium/genética , Mycobacterium/patogenicidade , Ácidos Micólicos/análise , Fenótipo , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Alinhamento de Sequência , Tuberculose dos Linfonodos/cirurgiaRESUMO
We evaluated the BDProbeTec ET system (Becton Dickinson, Sparks, Md.), a strand displacement amplification-based technique, for direct detection of Mycobacterium tuberculosis in 867 clinical samples. Of 294 extrapulmonary specimens, 52 had positive results by both BDProbeTec ET and culture and 209 had negative results by both methods; sensitivity and specificity were 76.5 and 95.9%, respectively. After resolution of discrepancies, the sensitivity rose to 77.8%.
Assuntos
Mycobacterium tuberculosis/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Kit de Reagentes para Diagnóstico , Tuberculose Pulmonar/microbiologia , Tuberculose/microbiologia , Líquidos Corporais/microbiologia , Meios de Cultura , DNA Bacteriano/análise , Humanos , Mycobacterium tuberculosis/genética , Sistema Respiratório/microbiologia , Sensibilidade e EspecificidadeRESUMO
This report describes detailed taxonomic and phylogenetic analysis of 15 non-tuberculous mycobacteria (NTMs) isolated from human pathological specimens in a Caribbean setting (12 slow-growers and three rapid-growers) that were not identified by cultural and biochemical tests and drug-susceptibility results. These isolates were further studied using PCR restriction fragment length polymorphism analysis (PRA) of a 441bp hsp65 fragment, as well as the sequencing of 16S rDNA and hsp65 DNA, and HPLC of the mycolic acids. Our results showed that taxonomic position of well-defined NTMs was resolved by PRA and sequencing of hsp65, nonetheless, it was not suitable to investigate rarely observed or new strains that required 16S rDNA sequencing and HPLC for a definite response. Unrooted neighbor-joining phylogenetic trees were drawn based upon the 16S rDNA and hsp65 sequences of the 15 NTMs compared with those from described species (73 for 16S rDNA and 45 for hsp65). For most of the NTMs not showing an exactly matching sequence with either hsp65 or 16S rDNA in the GenBank, the phylogenetic tree was able to provide with useful indications about their relatedness to known species. In such a case, a concording HPLC pattern with the sequence data and the place of the strain within the tree could lead to a potential identification. We also identified three identical isolates that define a new mycobacterial species within the group of M. simiae-related mycobacteria. The isolation and characterization of mycobacteria from new settings may lead to identify potential pathogens that may propogate in future because of increased human migration, travels, and climatic and ecological changes of the modern world.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Classificação , Mycobacterium/classificação , Filogenia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias/genética , Região do Caribe , Chaperonina 60 , Chaperoninas/genética , Criança , Cromatografia Líquida de Alta Pressão , DNA Ribossômico/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mycobacterium/genética , Mycobacterium/isolamento & purificação , Ácidos Micólicos/análise , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNARESUMO
The possibility that the strains included within the Mycobacterium avium complex (MAC), but not belonging either to M. avium or to Mycobacterium intracellulare, may be members of undescribed taxa, has already been questioned by several taxonomists. A very homogeneous cluster of 12 strains characterized by identical nucleotide sequences both in the 16S rDNA and in the 16S-23S internal transcribed spacer was investigated. Similar strains, previously reported in the literature, had been assigned either to the species M. intracellulare on the basis of the 16S rDNA similarity or to the group of MAC intermediates. However, several phenotypical and epidemiological characteristics seem to distinguish these strains from all other MAC organisms. The unique mycolic acid pattern obtained by HPLC is striking as it is characterized by two clusters of peaks, instead of the three presented by all other MAC organisms. All of the strains have been isolated from humans and all but one came from the respiratory tract of elderly people. The clinical significance of these strains, ascertained for seven patients, seems to suggest an unusually high virulence. The characteristics of all the strains reported in the literature, genotypically identical to the ones described here, seem to confirm our data, without reports of isolations from animals or the environment or, among humans, from AIDS patients. Therefore, an elevation of the MAC variant was proposed and characterized here, with the name Mycobacterium chimaera sp. nov.; this increases the number of species included in the M. avium complex. The type strain is FI-01069T (=CIP 107892T=DSM 44623T).