RESUMEN
Human papillomavirus (HPV) epidemiological and vaccine studies require highly sensitive HPV detection systems. The widely used broad-spectrum SPF10-DEIA-LiPA25 (SPF10 method) has reduced sensitivity toward HPV-45 and -59. Therefore, anogenital samples from the PASSYON study were retrospectively analyzed with type-specific (TS) HPV-45 and -59 real-time quantitative PCR (qPCR) assays. The SPF10 method missed 51.1% of HPV-45 and 76.1% of HPV-59 infections that were detected by the TS qPCR assays. The viral copy number (VCn) of SPF10-missed HPV-45 and -59 was significantly lower than SPF10-detected HPV-45 and -59 (P < 0.0001 for both HPV types). Sanger sequencing showed no phylogenetic distinction between SPF10-missed and SPF10-detected HPV-59 variants, but variants bearing the A6562G single-nucleotide polymorphism (SNP) in the SPF10 target region were more likely to be missed (P = 0.0392). HPV cooccurrence slightly influenced the detection probability of HPV-45 and -59 with the SPF10 method. Moreover, HPV-59 detection with the SPF10 method was hampered more in nonvaccinated women than vaccinated women, likely due to a stronger masking effect by increased HPV cooccurrence in the former group. Consequently, the SPF10 method led to a strong negative vaccine effectiveness (VE) of -84.6% against HPV-59, while the VE based on TS qPCR was 3.1%. For HPV-45, the relative increase in detection in nonvaccinated women compared vaccinated women was more similar, resulting in comparable VE estimates. In conclusion, this study shows that HPV-45 and -59 detection with the SPF10 method is dependent on factors including VCn, HPV cooccurrence, and vaccination, thereby showing that knowledge of the limitations of the HPV detection method used is of great importance.
Asunto(s)
Infecciones por Papillomavirus , Vacunas , ADN Viral , Femenino , Humanos , Papillomaviridae/genética , Infecciones por Papillomavirus/diagnóstico , Reacción en Cadena en Tiempo Real de la Polimerasa , Estudios RetrospectivosRESUMEN
Patients infected by Mycoplasma genitalium are often treated empirically with the macrolide azithromycin. Macrolide resistance is becoming quite common; empirical treatment is compromised. Sequencing was initially used to detected azithromycin resistance-associated mutations. As this was laborious, qPCRs have been developed for their detection. In the present study, we describe a fast, sensitive, and specific qPCR assay that enables routine testing of M. genitalium and macrolide resistance-associated mutations in a single assay. M. genitalium positive clinical samples were used to compare (i) the commonly used MgPa assay for the detection of M. genitalium infections (MgPa qPCR), (ii) a combined 23S rRNA gene PCR/sequencing assay (Mg23S qPCR/Sequencing) to identify macrolide resistance-associated mutations, and (iii) our newly developed probe-based melt curve qPCR for simultaneous detection of M. genitalium and macrolide resistance-associated mutations (Macrolide-R/MG ELITe MGB Kit, Elitech Bothel USA in short Mg MacrolideR qPCR). Specificity of the qPCR was tested using urogenital samples that were tested positive for a range of other micro-organisms. M. genitalium was detected in 196/236 (83.1%) samples by the MgPa qPCR, versus 172/236 (72.9%) by the combined Mg23S qPCR/Sequencing, and 202/236 (85.6%) by the Mg MacrolideR qPCR. The Mg MacrolideR qPCR showed high concordance to the Mg23S qPCR/Sequencing assay (201 vs 202 could be genotyped, respectively) for the detection of the macrolide resistant mutations. None of the other urogenital pathogens were tested positive in the Mg MacrolideR qPCR, indicating specificity. The Mg MacrolideR qPCR is fast, sensitive, specific, and can easily be implemented in the routine diagnostics.
Asunto(s)
Antituberculosos/farmacología , Farmacorresistencia Bacteriana , Macrólidos/farmacología , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/microbiología , Mycoplasma genitalium/efectos de los fármacos , Mycoplasma genitalium/genética , Adolescente , Adulto , Antituberculosos/uso terapéutico , Femenino , Humanos , Macrólidos/uso terapéutico , Masculino , Persona de Mediana Edad , Mutación , Infecciones por Mycoplasma/tratamiento farmacológico , ARN Ribosómico 23S/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Sensibilidad y Especificidad , Adulto JovenRESUMEN
OBJECTIVES: The association between mupirocin use and plasmid-based high-level resistance development mediated through mupA in CoNS has not been quantified. We determined acquisition of mupirocin resistance in Staphylococcus aureus and CoNS in surgery patients treated peri-operatively with mupirocin. PATIENTS AND METHODS: Patients admitted for surgery were treated with nasal mupirocin ointment and chlorhexidine soap for 5 days, irrespective of S. aureus carrier status. Nasal swabs were obtained before decolonization (T1) and 4 days after surgery (T2) and were inoculated onto agars containing 8 mg/L mupirocin. Staphylococci were identified by MALDI-TOF MS and mupirocin resistance was confirmed by Etest. RESULTS: Among 1578 surgical patients, 936 (59%) had nasal swabs obtained at T1 and T2; 192 (21%) patients carried mupirocin-resistant CoNS at T1 and 406 (43%) at T2 (P<0.001). Of 744 patients not colonized at T1, 277 acquired resistance (37%), corresponding to an acquisition rate of 7.4/100 patient days at risk. In all, 588 (97%) of 607 mupirocin-resistant CoNS had an MIC >256 mg/L (high level) and 381 of 383 (99.5%) were mupA positive. No acquisition of mupirocin resistance was observed in S. aureus. CONCLUSIONS: Acquisition of mupirocin resistance following decolonization was widespread in CoNS and absent in S. aureus. As almost all isolates harboured the mupA gene, monitoring resistance development in S. aureus when decolonization strategies containing mupirocin are used is recommended.
Asunto(s)
Farmacorresistencia Bacteriana , Mupirocina/farmacología , Mupirocina/uso terapéutico , Cavidad Nasal/microbiología , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus/efectos de los fármacos , Estudios de Cohortes , Humanos , Pruebas de Sensibilidad Microbiana , Plásmidos , Estudios Prospectivos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Staphylococcus/clasificación , Staphylococcus/aislamiento & purificaciónAsunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana/genética , Macrólidos/farmacología , Infecciones por Mycoplasma/microbiología , Mycoplasma genitalium/efectos de los fármacos , Mycoplasma genitalium/genética , ARN Ribosómico 23S/genética , Enfermedades Bacterianas de Transmisión Sexual/microbiología , Adolescente , Adulto , Anciano , Antibacterianos/uso terapéutico , Azitromicina/uso terapéutico , Femenino , Humanos , Macrólidos/uso terapéutico , Masculino , Persona de Mediana Edad , Mutación , Infecciones por Mycoplasma/tratamiento farmacológico , Infecciones por Mycoplasma/epidemiología , Mycoplasma genitalium/aislamiento & purificación , Países Bajos/epidemiología , Prevalencia , Enfermedades Bacterianas de Transmisión Sexual/tratamiento farmacológico , Enfermedades Bacterianas de Transmisión Sexual/epidemiologíaRESUMEN
A 13-year old neutropenic boy succumbed to bacteremia and sepsis with a Pseudomonas aeruginosa strain that rapidly developed resistance to carbapenems during meropenem monotherapy. Whole genome sequencing of the susceptible and resistant blood culture isolates revealed the meropenem-resistant phenotype to be caused by truncation of the OprD gene, which added to a preexisting inactivated mexR gene.
Asunto(s)
Antibacterianos/administración & dosificación , Meropenem/administración & dosificación , Mutación , Porinas/genética , Infecciones por Pseudomonas/tratamiento farmacológico , Pseudomonas aeruginosa/efectos de los fármacos , Resistencia betalactámica , Adolescente , Bacteriemia/tratamiento farmacológico , Bacteriemia/microbiología , Cultivo de Sangre , Resultado Fatal , Humanos , Masculino , Infecciones por Pseudomonas/microbiología , Pseudomonas aeruginosa/genética , Eliminación de Secuencia , Secuenciación Completa del GenomaRESUMEN
INTRODUCTION: The aim of this study was to determine the prevalence of Mycoplasma genitalium infection and the resistance to macrolides within a general population in Madrid in 2015. METHODS: We collected 359 urine samples from a general population with symptoms of sexually transmitted infections (STIs). All samples underwent a real-time PCR. For the detection of macrolide resistance, a 283bp fragment of region V of the 23S rRNA gene of M. genitalium was amplified and sequenced. RESULTS: We found a prevalence of 3.34% of M. genitalium and a macrolide resistance rate of 20%. In males, the prevalence was 6.62% and in women 0.96%, being significantly higher in males. CONCLUSIONS: The prevalence obtained shows that it is a pathogen to consider in our environment. These findings stress the need for routine testing of M. genitalium infections and would seem to suggest the advisability of resistance testing.
Asunto(s)
Antibacterianos/farmacología , Enfermedades de los Genitales Femeninos/epidemiología , Enfermedades de los Genitales Femeninos/microbiología , Enfermedades de los Genitales Masculinos/epidemiología , Enfermedades de los Genitales Masculinos/microbiología , Macrólidos/farmacología , Infecciones por Mycoplasma/epidemiología , Infecciones por Mycoplasma/microbiología , Mycoplasma genitalium/efectos de los fármacos , Adulto , Antibacterianos/uso terapéutico , Farmacorresistencia Bacteriana , Femenino , Enfermedades de los Genitales Femeninos/tratamiento farmacológico , Enfermedades de los Genitales Masculinos/tratamiento farmacológico , Humanos , Macrólidos/uso terapéutico , Masculino , Infecciones por Mycoplasma/tratamiento farmacológico , Prevalencia , Adulto JovenRESUMEN
Campylobacter, Arcobacter, and Helicobacter species have been isolated from many vertebrate hosts, including birds, mammals, and reptiles. Multiple studies have focused on the prevalence of these Epsilonproteobacteria genera in avian and mammalian species. However, little focus has been given to the presence within reptiles, and their potential zoonotic and pathogenic roles. In this study, occurrence, diversity, and host association of intestinal Epsilonproteobacteria were determined for a large variety of reptiles. From 2011 to 2013, 444 cloacal swabs and fecal samples originating from 417 predominantly captive-held reptiles were screened for Epsilonproteobacteria. Campylobacter, Arcobacter, and Helicobacter genus specific PCRs were performed directly on all samples. All samples were also cultured on selective media and screened for the presence of Epsilonproteobacteria. Using a tiered approach of AFLP, atpA, and 16S rRNA sequencing, 432 Epsilonproteobacteria isolates were characterized at the species level. Based on PCR, Campylobacter, Arcobacter, and Helicobacter were detected in 69.3% of the reptiles; 82.5% of the chelonians, 63.8% of the lizards, and 58.0% of the snakes were positive for one or more of these genera. Epsilonproteobacteria were isolated from 22.1% of the reptiles and were isolated most frequently from chelonians (37.0%), followed by lizards (19.6%) and snakes (3.0%). The most commonly isolated taxa were Arcobacter butzleri, Arcobacter skirrowii, reptile-associated Campylobacter fetus subsp. testudinum, and a putative novel Campylobacter taxon. Furthermore, a clade of seven related putative novel Helicobacter taxa was isolated from lizards and chelonians. This study shows that reptiles carry various intestinal Epsilonproteobacteria taxa, including several putative novel taxa.
Asunto(s)
Arcobacter/aislamiento & purificación , Campylobacter/aislamiento & purificación , Helicobacter/aislamiento & purificación , Reptiles/microbiología , Animales , Arcobacter/clasificación , Arcobacter/genética , Campylobacter/clasificación , Campylobacter/genética , Variación Genética , Helicobacter/clasificación , Helicobacter/genética , Intestinos/microbiología , FilogeniaRESUMEN
Introduction: The aim of this study was to determine the prevalence of Mycoplasma genitalium infection and the resistance to macrolides within a general population in Madrid in 2015. Methods: We collected 359 urine samples from a general population with symptoms of sexually transmitted infections (STIs). All samples underwent a real-time PCR. For the detection of macrolide resistance, a 283bp fragment of region V of the 23S rRNA gene of M. genitaliumwas amplified and sequenced. Results: We found a prevalence of 3.34% of M. genitalium and a macrolide resistance rate of 20%. In males, the prevalence was 6.62% and in women 0.96%, being significantly higher in males. Conclusions: The prevalence obtained shows that it is a pathogen to consider in our environment. These findings stress the need for routine testing of M. genitalium infections and would seem to suggest the advisability of resistance testing (AU)
Introducción: El objetivo de este estudio fue determinar en 2015 en una población general de Madrid la prevalencia de infección por Mycoplasma genitalium (M. genitalium) y la resistencia a macrólidos. Métodos: Se recogieron 359 muestras de orina procedentes de una población general con síntomas de infección de transmisión sexual. A todas las muestras se les realizó una PCR a tiempo real. Para la detección de resistencias a macrólidos, se amplificó y secuenció un fragmento de 283 pb de la región V del gen 23S rRNA de M. genitalium. Resultados: Se encontró una prevalencia de un 3,34% de M. genitalium y un 20% de resistencia a macrólidos. En varones la prevalencia fue del 6,62% y en mujeres del 0,96%, siendo significativamente superior en varones. Conclusión: La prevalencia obtenida muestra que es un patógeno a considerar en nuestro entorno. Estos hallazgos hacen hincapié en la necesidad de realizar pruebas de rutina de infección por M. genitalium y argumentan que es recomendable realizar pruebas de resistencia (AU)