RESUMO
OBJECTIVES: A large clonal outbreak of multidrug-resistant CC17 ST17 Enterococcus faecium containing Tn5382 in a hospital in the north of Spain is described. METHODS: We characterized vancomycin-resistant E. faecium isolates from 10 infected and 40 colonized inpatients from a single hospital by PFGE, multiple-locus variable-number tandem-repeat analysis (MLVA) and multilocus sequence typing (MLST). Genes encoding antibiotic resistance (ampicillin, aminoglycosides, macrolides, quinupristin/dalfopristin, quinolones, tetracycline) and putative virulence traits were analysed. RESULTS: All isolates showed highly similar PFGE profiles and were assigned to the type MT1 by MLVA and to ST17 (CC17) by MLST. The Tn5382 type identified in all isolates was linked to pbp5 and contained a 5 bp deletion and 10 point mutations within the intergenic vanS(B)-vanY(B) region. Other resistance genes identified were erm(B), mef(E), tet(M), ant(6')-Ia, aph(3')-IIIa and aac(6')-Ie-aph(2'')-Ia. All isolates carried the unexpressed tet(M) gene. The high level of ciprofloxacin resistance was attributable to the first described Gly-61 and Ile-80 mutations in ParC and the Tyr-83 or Arg-83 mutations in GyrA. All isolates contained esp. The presence of hyl was variable. CONCLUSIONS: A large clonal outbreak caused by multidrug-resistant CC17 E. faecium containing pbp5-Tn5382 is described. The persistence of this clone, which has been recovered from both hospital and community settings since 2005, and the possibility of transferring this Tn5382 to other epidemic ampicillin-resistant clonal types currently circulating in Spain might contribute to increasing the prevalence of vancomycin-resistant enterococci in our area. This study constitutes the first description of mef(E) in E. faecium.
Assuntos
Elementos de DNA Transponíveis , Farmacorresistência Bacteriana Múltipla , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/isolamento & purificação , Infecções por Bactérias Gram-Positivas/epidemiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Técnicas de Tipagem Bacteriana , Portador Sadio/microbiologia , Análise por Conglomerados , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Impressões Digitais de DNA , DNA Bacteriano/química , DNA Bacteriano/genética , Enterococcus faecium/classificação , Genes Bacterianos , Genótipo , Hospitais , Humanos , Dados de Sequência Molecular , Análise de Sequência de DNA , Espanha/epidemiologia , Fatores de Virulência/genéticaRESUMO
BACKGROUND AND OBJECTIVE: Our goal was to establish the different therapeutic regimens used in our clinical setting, and to determine the prevalence of genotypic resistances in patients under antiretroviral therapy, analyzing the relationship between the appearance of mutations and treatments along with other HIV related variables. MATERIAL AND METHOD: 191 samples from the same number of patients who were on antiretroviral therapy and virological failure were analyzed. Samples were processed by means of the genotypic technique LiPA in order to study the presence of mutations in the reverse transcriptase (RT) and the protease (P) genes. Prescribed therapeutic regimens and epidemiological variables relevant in HIV infection were also analyzed. RESULTS: Overall resistance prevalence was 72.32%. By LiPA, RT mutations were detected in 71.43% of patients, being M184V, T215Y and L41M the most frequent ones. Moreover, P mutations were detected in 59.38% of cases, being V82A, L90M and I84V the most frequent ones. 61.02% of the patients presented one or more mutations against the reverse transcriptase inhibitors included in their treatment. With regard to protease inhibitors, this fact was documented in 28.81% of cases, and in 23.73% of patients receiving both reverse transcriptase inhibitors and protease inhibitors. CONCLUSIONS: Although the analysis of the mutation patterns by LiPA has known limitations, the prevalence of resistances in our study was different from that reported by other authors, being lower in the P gene and higher in the RT one. Of note, a high proportion of patients showed mutations against the drugs included in their prescribed treatment.
Assuntos
Infecções por HIV/tratamento farmacológico , HIV/efeitos dos fármacos , HIV/genética , Farmacorresistência Viral/genética , Feminino , Genótipo , Humanos , MasculinoRESUMO
FUNDAMENTO Y OBJETIVO: Describir las pautas de tratamiento empleadas en pacientes infectadospor el virus de la inmunodeficiencia humana (VIH) de nuestro entorno asistencial y determinarla prevalencia de resistencias en pacientes en tratamiento antirretroviral, analizando la relaciónentre la aparición de mutaciones con los regímenes terapéuticos junto con otras variables deinterés.MATERIAL Y MÉTODO: Se analizaron 191 muestras de pacientes en fracaso virológico que estabanrecibiendo tratamiento antirretroviral, mediante la técnica de genotipificación LiPA para determinarla presencia de mutaciones en el gen de la retrotranscriptasa (RT) y de la proteasa (P).Se analizaron las pautas de tratamiento recibido, así como las variables relevantes en la infecciónpor el VIH.RESULTADOS: La prevalencia global de resistencias fue del 72,32%. Mediante LiPA se detectaronmutaciones en RT en el 71,43%, y las más frecuentes fueron M184V, T215Y y L41M. La prevalenciade mutaciones en P fue del 53,38%, siendo las V82A, L90M e I84V más frecuentesrespecto a las mutaciones frente a los fármacos incluidos en la pauta de tratamiento de los pacientes.El 61,02% presentaba una o varias frente a los inhibidores de la transcriptasa inversa,el 28,81% para los inhibidores de la proteasa y el 23,73% tanto frente a los primeros como alos segundos.Se constataron 72 combinaciones terapéuticas diferentes; el tratamiento triple fue la modalidadprescrita en el 73,46% de los casos.CONCLUSIONES: Aun con las limitaciones inherentes a la técnica de LiPA, la prevalencia de resistenciasen nuestro estudio fue diferente de la comunicada por otros autores, siendo menor enel gen P y mayor en TI. Una alta proporción de pacientes mostraron mutaciones frente a los fármacosincluidos en su tratamiento
BACKGROUND AND OBJECTIVE: Our goal was to establish the different therapeutic regimens used inour clinical setting, and to determine the prevalence of genotypic resistances in patients underantiretroviral therapy, analyzing the relationship between the appearance of mutations and treatmentsalong with other HIV related variables.MATERIAL AND METHOD: 191 samples from the same number of patients who were on antiretroviraltherapy and virological failure were analyzed. Samples were processed by means of the genotypictechnique LiPA in order to study the presence of mutations in the reverse transcriptase (RT)and the protease (P) genes. Prescribed therapeutic regimens and epidemiological variables relevantin HIV infection were also analyzed.RESULTS: Overall resistance prevalence was 72.32%. By LiPA, RT mutations were detected in71.43% of patients, being M184V, T215Y and L41M the most frequent ones. Moreover, P mutationswere detected in 59.38% of cases, being V82A, L90M and I84V the most frequentones. 61.02% of the patients presented one or more mutations against the reverse transcriptaseinhibitors included in their treatment. With regard to protease inhibitors, this fact was documentedin 28.81% of cases, and in 23.73% of patients receiving both reverse transcriptase inhibitorsand protease inhibitors.CONCLUSIONS: Although the analysis of the mutation patterns by LiPA has known limitations, theprevalence of resistances in our study was different from that reported by other authors, beinglower in the P gene and higher in the RT one. Of note, a high proportion of patients showedmutations against the drugs included in their prescribed treatment
Assuntos
Masculino , Feminino , Humanos , Infecções por HIV/tratamento farmacológico , HIV , HIV/genética , Farmacorresistência Viral/genética , GenótipoRESUMO
BACKGROUND: Genotyping testing has been accepted as a guidance in the therapeutic management of Human Immunodeficiency virus 1 (HIV-1). However, optimization of the available routine techniques for such purpose has not been fulfilled. OBJECTIVE: To evaluate the use of three RNA extraction methods in order to be applied in the genotypic HIV-1 resistance testing by LiPA. STUDY DESIGN: Comparative prospective study of three HIV-1 RNA extraction methods. Forty-eight plasma samples were tested for the determination of viral load (VL) by means of Cobas Amplicor HIV-1 Monitor (Roche Diagnostics. Branchburg, NJ, USA), preserving the obtained RNA extracts. RNA was also extracted using two other techniques: "SV Total RNA Isolation System" (Promega Corporation. Madison, WI, USA) and "QIAamp Viral RNA" (QIAGEN Inc., Valencia, CA, USA). The three RNA extracts were processed in parallel for the detection of HIV resistance by LiPA, and bands were recorded comparatively. RESULTS: Results obtained by Roche extraction method were superior, followed by those of Qiagen and Promega, in the several studied parameters. First, proportion of amplified samples (75.0% by Promega versus 95.8 by Qiagen and 97.9% by Roche for LiPA RT and 97.7% by Promega versus 100.0% by Roche and Qiagen for LiPA P); second, percentage of combined mutations patterns, and third, differences in band intensity. Thus, for LiPA RT 51.4% and 54.3% of the samples showed greater intensity after Roche and Qiagen extractions, respectively. These percentages dropped to 12.8 and 19.1 for LiPA P. CONCLUSIONS: The outcome obtained by LiPA after RNA extraction by Roche methodology was remarkably superior to those of Promega and Qiagen. LiPA technique needs further optimization, especially the sample amplification phase of LiPA RT.
Assuntos
Fármacos Anti-HIV/farmacologia , HIV-1/efeitos dos fármacos , RNA Viral/isolamento & purificação , Farmacorresistência Viral/genética , Genótipo , Inibidores da Protease de HIV/farmacologia , HIV-1/genética , Humanos , Reação em Cadeia da Polimerase/métodos , RNA Viral/genética , Inibidores da Transcriptase Reversa/farmacologiaRESUMO
BACKGROUND: The aim of the present study is to asses the prevalence of resistances in a group of patients with virological failure establishing the relationship between the appearance of mutations and the given antiretroviral therapy along with other variables used in these patients follow-up. PATIENTS AND METHOD: Samples belonging to 88 patients were selected either with viral load levels above 30.000 copies/ml after reaching undetectable viral load levels, or with persistently detectable levels above 1.000 copies/ml. Resistances were tested by means of Line Probe Assay (LiPA). The history of patients' antiretroviral treatments was reviewed. RESULTS: Mutations were observed in 52,6% of cases for reverse transcriptase (RT) an in 81,8% for the protease genes, being T215Y and V82A the most frequently detected ones. Mutations coferring resistance to the given antiretrovirals appeared in 33 cases. No statistical significance was observed between the presence of mutations and the administered therapy. In the multivariate analysis we found for LiPA RT a greater risk of appearance of mutations according to patient motility (OR = 4,0). CONCLUSIONS: the prevalence of resistance mutations in patients with virologic failure is placed around 50% in both genes. A consensus in the definition of virologic failure in HIV infected patients is urged.
Assuntos
Terapia Antirretroviral de Alta Atividade , Soropositividade para HIV , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Farmacorresistência Viral , Endopeptidases/genética , Feminino , Genótipo , Soropositividade para HIV/tratamento farmacológico , Soropositividade para HIV/epidemiologia , Soropositividade para HIV/genética , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Falha de TratamentoRESUMO
FUNDAMENTO: El objetivo del presente estudio es establecer la prevalencia de resistencias en un grupo de pacientes en los que el tratamiento antirretroviral parece haber fracasado, analizando la relación entre la aparición de mutaciones y los regímenes terapéuticos seguidos, junto con otras variables asociadas al seguimiento. PACIENTES Y MÉTODO: Se seleccionaron muestras correspondientes a 88 pacientes con cargas virales superiores a 30.000 copias ARN/ml tras haber alcanzado valores indetectables, o cargas virales persistentemente detectables superiores a 1.000 copias ARN/ml. Se analizaron las resistencias mediante Line Probe Assay (LiPA) y se revisó la historia de tratamientos antirretrovirales de los pacientes. RESULTADOS: Se obtuvieron mutaciones en el 52,6 por ciento de los casos para el gen de la transcriptasa inversa (RT) y en un 51,8 por ciento para el de la proteasa, siendo la T215Y y la V82A las más frecuentes. En 33 casos hubo coincidencia entre la aparición de mutaciones que conferían resistencia para los fármacos que los pacientes recibían. No se encontró significación estadística entre la aparición de mutaciones y los tratamientos empleados en el seguimiento de los pacientes. En el análisis multivariante se encontró para LiPA RT, respecto a la movilidad, un riesgo 4,0 veces superior de aparición de mutaciones. CONCLUSIONES: La prevalencia de mutaciones de resistencia en pacientes en fracaso virológico se sitúa en torno al 50 por ciento de los casos en ambos genes. Es necesario establecer un consenso para definir el fracaso virológico en los pacientes con infección por el VIH. (AU)
Assuntos
Pessoa de Meia-Idade , Criança , Pré-Escolar , Adolescente , Adulto , Idoso , Masculino , Feminino , Humanos , Terapia Antirretroviral de Alta Atividade , Soropositividade para HIV , Prevalência , Falha de Tratamento , Endopeptidases , Cuidados Pós-Operatórios , Estudos Retrospectivos , Prognóstico , Farmacorresistência Viral , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Glicemia , Insulinoma , Genótipo , Neoplasias PancreáticasRESUMO
BACKGROUND: To date, antiretroviral therapies have not led to total HIV eradication in infected individuals. The achievement of undetectable viremia levels has been regarded as essential when monitorizing the efficacy of these therapies. The objectives of our study were to assess the time needed to achieve viral load (VL) undetectable levels and subsequent VL risings, as well as to determine the clinical variables associated with both events. PATIENTS AND METHOD: Owing to a follow-up period greater than 1,000 days, 314 patients were retrospectively selected. We determined the percentages of patients with undetectable VL levels and of those with subsequent VL rising. Times needed to reach both events were calculated by Kaplan-Meier analysis. RESULTS: 182 individuals (58%; 95% CI, 52.3-63.5) reached undetectable VL levels with a median follow-up time of 862 days. A more regular VL quantitation was associated with such event. The VL level of 38 out of these individuals (20.9%; 95% CI, 15.2-27.5) increased subsequently (> 30,000 RNA copies/ml) with a mean time of 328 days (after the first undetectable VL determination); this event was associated with changes in the clinical centre where the patient was being cared for. CONCLUSIONS: In our series, the proportion of individuals with undetectable VL levels and therapeutic failure was low. The frequency of VL determination was a determining factor in the evolution of these patients.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/tratamento farmacológico , Infecções por HIV/virologia , Carga Viral , Adulto , Distribuição de Qui-Quadrado , Estudos de Coortes , Feminino , Seguimentos , Infecções por HIV/transmissão , HIV-1/genética , Humanos , Masculino , RNA Viral/análise , Estudos Retrospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Risco , Abuso de Substâncias por Via Intravenosa/complicações , Análise de Sobrevida , Fatores de TempoRESUMO
We evaluated two methods from Roche and Promega for RNA extraction prior to the genotypic detection of human immunodeficiency virus type 1 resistance by line probe assay (LiPA). Fifty plasma RNA extracts were processed in parallel by LiPA. Results obtained by the Roche method were superior in the proportion of amplified samples, the percentage of mutated samples, and band intensity.
Assuntos
Fármacos Anti-HIV/farmacologia , Farmacorresistência Viral , HIV-1/efeitos dos fármacos , Técnicas de Sonda Molecular , RNA Viral/isolamento & purificação , Fármacos Anti-HIV/uso terapêutico , Genótipo , Infecções por HIV/tratamento farmacológico , Infecções por HIV/virologia , Protease de HIV/genética , Transcriptase Reversa do HIV/genética , HIV-1/genética , HumanosRESUMO
FUNDAMENTO: Los tratamientos antirretrovirales no han logrado la erradicación total del virus de la inmunodeficiencia humana (VIH) de los individuos infectados hasta el momento actual. La consecución de niveles indetectables de viremia se ha señalado como esencial a la hora de delimitar la eficacia de los fármacos. El objetivo de nuestro estudio fue determinar tanto la proporción de individuos como el tiempo real para alcanzar carga viral (CV) no detectable y para documentar una elevación posterior, así como las variables clínicas que se asocian a ambas circunstancias. PACIENTES Y MÉTODO: Se seleccionó retrospectivamente a 314 pacientes en función de la monitorización de CV durante un tiempo superior a 1.000 días. Se describieron los porcentajes de aquellos pacientes que alcanzaron CV 'no detectables' y de aquellos en los que posteriormente se documentó una elevación (> 30.000 copias ARN/ml). Asimismo se describió el tiempo necesario para ambas circunstancias y se realizó un análisis de Kaplan-Meier. RESULTADOS: Alcanzaron CV 'no detectables' 182 pacientes (58 per cent; intervalo de confianza [IC] del 95 per cent, 52,3-63,5) con una mediana de tiempo de 862 días. Con estas viremias se asoció una frecuencia de cuantificación de CV más regular. En 38 de estos individuos (20,9 per cent; IC del 95 per cent, 15,2-27,5), la CV experimentó una elevación posterior en un tiempo medio de 328 días; esta circunstancia se asoció a cambios del centro asistencial. CONCLUSIONES: La proporción de individuos infectados por el VIH que alcanzaron el éxito y fracasaron desde el punto de vista terapéutico fue baja y la frecuencia de cuantificación de la CV fue determinante en su evolución (AU)
