RESUMEN
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.
Asunto(s)
Anticuerpos Antivirales/sangre , Dengue/diagnóstico , Técnica del Anticuerpo Fluorescente , Infección por el Virus Zika/diagnóstico , Antígenos Virales/inmunología , Reacciones Cruzadas , Dengue/inmunología , Virus del Dengue , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Sensibilidad y Especificidad , Pruebas Serológicas , Virus Zika , Infección por el Virus Zika/inmunologíaAsunto(s)
Personal de Laboratorio Clínico/psicología , Seguridad del Paciente/legislación & jurisprudencia , Gestión de Riesgos/ética , Carga de Trabajo/psicología , Lugar de Trabajo/normas , Patógenos Transmitidos por la Sangre/aislamiento & purificación , Infección Hospitalaria/prevención & control , Pruebas Diagnósticas de Rutina/ética , Pruebas Diagnósticas de Rutina/estadística & datos numéricos , Hepatitis B/sangre , Hepatitis B/diagnóstico , Antígenos de la Hepatitis B/sangre , Antígenos de la Hepatitis B/genética , Antígenos de la Hepatitis B/inmunología , Humanos , Inmunoglobulina M/sangre , Inmunoglobulina M/inmunología , Italia/epidemiología , Personal de Laboratorio Clínico/economía , Personal de Laboratorio Clínico/normas , Aislamiento de Pacientes/normas , Seguridad del Paciente/normas , Diálisis Renal/métodos , Lugar de Trabajo/estadística & datos numéricosRESUMEN
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.
Asunto(s)
Enfermedades Pulmonares/diagnóstico , Enfermedades Pulmonares/microbiología , Mycobacterium/clasificación , Mycobacterium/aislamiento & purificación , Proteínas Bacterianas/genética , Técnicas de Tipificación Bacteriana , Chaperonina 60 , Chaperoninas/genética , ADN Bacteriano/análisis , ADN Espaciador Ribosómico/análisis , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Mycobacterium/genética , Infecciones por Mycobacterium/diagnóstico , Infecciones por Mycobacterium/microbiología , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Análisis de Secuencia de ADNRESUMEN
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).
Asunto(s)
Infecciones por Mycobacterium no Tuberculosas/microbiología , Ácidos Micólicos/análisis , Micobacterias no Tuberculosas/clasificación , Micobacterias no Tuberculosas/aislamiento & purificación , Adulto , Anciano de 80 o más Años , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Líquido del Lavado Bronquioalveolar/microbiología , Chaperonina 60 , Chaperoninas/genética , Niño , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Femenino , Genes de ARNr , Alemania , Humanos , Italia , Masculino , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Micobacterias no Tuberculosas/fisiología , Filogenia , Pigmentos Biológicos , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , Esputo/microbiología , Infección de Heridas/microbiologíaRESUMEN
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.
Asunto(s)
Antituberculosos/uso terapéutico , Discitis/tratamiento farmacológico , Discitis/microbiología , Mycobacterium/efectos de los fármacos , Mycobacterium/aislamiento & purificación , Secuencia de Bases , ADN Espaciador Ribosómico/análisis , Genes de ARNr , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Mycobacterium/clasificación , Mycobacterium/genética , Infecciones por Mycobacterium/tratamiento farmacológico , Infecciones por Mycobacterium/microbiología , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Análisis de Secuencia de ADN , Resultado del TratamientoRESUMEN
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.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Infecciones por Mycobacterium/microbiología , Mycobacterium/aislamiento & purificación , Infecciones Oportunistas/microbiología , Adulto , Anciano , Humanos , Masculino , Persona de Mediana Edad , Mycobacterium/clasificación , Mycobacterium/genéticaRESUMEN
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.
Asunto(s)
Ganglios Linfáticos/microbiología , Mycobacterium/clasificación , Mycobacterium/aislamiento & purificación , Esputo/microbiología , Adulto , Anciano , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Pared Celular/química , Chaperonina 60 , Chaperoninas/genética , Niño , Cromatografía de Gases , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Espaciador Ribosómico , Farmacorresistencia Bacteriana , Heces/microbiología , Femenino , Genes de ARNr , Humanos , Lípidos/análisis , Lípidos/aislamiento & purificación , Masculino , Datos de Secuencia Molecular , Mycobacterium/citología , Mycobacterium/fisiología , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
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.
Asunto(s)
Enfermedades Pulmonares/microbiología , Infecciones por Mycobacterium/microbiología , Mycobacterium , Anciano , Antibacterianos/uso terapéutico , Secuencia de Bases , ADN Bacteriano/análisis , Femenino , Humanos , Inmunocompetencia , Persona de Mediana Edad , Datos de Secuencia Molecular , Mycobacterium/clasificación , Infecciones por Mycobacterium/tratamiento farmacológico , Infecciones por Mycobacterium/inmunología , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
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.
Asunto(s)
Técnicas de Tipificación Bacteriana/métodos , Clasificación , Mycobacterium/clasificación , Filogenia , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Proteínas Bacterianas/genética , Región del Caribe , Chaperonina 60 , Chaperoninas/genética , Niño , Cromatografía Líquida de Alta Presión , ADN Ribosómico/análisis , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mycobacterium/genética , Mycobacterium/aislamiento & purificación , Ácidos Micólicos/análisis , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Análisis de Secuencia de ADNRESUMEN
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.
Asunto(s)
Mycobacterium/aislamiento & purificación , Tuberculosis Ganglionar/diagnóstico , Secuencia de Bases , Pared Celular/química , Cromatografía Líquida de Alta Presión , Femenino , Humanos , Escisión del Ganglio Linfático , Persona de Mediana Edad , Mycobacterium/genética , Mycobacterium/patogenicidad , Ácidos Micólicos/análisis , Fenotipo , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Alineación de Secuencia , Tuberculosis Ganglionar/cirugíaRESUMEN
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).
Asunto(s)
Complejo Mycobacterium avium/clasificación , Complejo Mycobacterium avium/genética , Anciano , Dermatoglifia del ADN , Elementos Transponibles de ADN , ADN Bacteriano/química , ADN Ribosómico/química , ADN Espaciador Ribosómico/química , Femenino , Genes de ARNr , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Datos de Secuencia Molecular , Complejo Mycobacterium avium/química , Complejo Mycobacterium avium/aislamiento & purificación , Infección por Mycobacterium avium-intracellulare/microbiología , Ácidos Micólicos/análisis , Hibridación de Ácido Nucleico , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
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%.
Asunto(s)
Sondas de ADN , Mycobacterium/aislamiento & purificación , Hibridación de Ácido Nucleico , Mycobacterium/clasificación , Sensibilidad y EspecificidadRESUMEN
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%.