Y155H amino acid substitution in influenza A(H1N1)pdm09 viruses does not confer a phenotype of reduced susceptibility to neuraminidase inhibitors.

The Y155H amino acid substitution in the neuraminidase gene (NA) has previously been associated with highly reduced inhibition by neuraminidase inhibitors in the seasonal H1N1 influenza A virus which circulated in humans before the 2009 pandemic. During the 2012/13 epidemic season in Spain, two A(H1N1) pdm09 viruses bearing the specific Y155H substitution in the NA were detected and isolated from two patients diagnosed with severe respiratory syndrome and pneumonia requiring admission to the intensive care unit. Contrary to what was observed in the seasonal A(H1N1) viruses, neither of the Y155H A(H1N1) pdm09 viruses described here showed a phenotype of reduced inhibition by NAIs as determined by the neuraminidase enzyme inhibition assay (MUNANA). High-throughput sequencing of the NA of both Y155H viruses showed that they were composed to >99% of H155 variants. We believe that this report can contribute to a better understanding of the biological significance of amino acid substitutions in the neuraminidase protein with regard to susceptibility of influenza viruses to neuraminidase inhibitors. This is of critical importance for optimal management of influenza disease patients.

Viral infections of the central nervous system in Spain: a prospective study.

The aim of the study was to determine the incidence of viruses causing aseptic meningitis, meningoencephalitis, and encephalitis in Spain. This was a prospective study, in collaboration with 17 Spanish hospitals, including 581 cases (CSF from all and sera from 280): meningitis (340), meningoencephalitis (91), encephalitis (76), febrile syndrome (7), other neurological disorders (32), and 35 cases without clinical information. CSF were assayed by PCR for enterovirus (EV), herpesvirus (herpes simplex [HSV], varicella-zoster [VZV], cytomegalovirus [CMV], Epstein-Barr [EBV], and human herpes virus-6 [HHV-6]), mumps (MV), Toscana virus (TOSV), adenovirus (HAdV), lymphocytic choriomeningitis virus (LCMV), West Nile virus (WNV), and rabies. Serology was undertaken when methodology was available. Amongst meningitis cases, 57.1% were characterized; EV was the most frequent (76.8%), followed by VZV (10.3%) and HSV (3.1%; HSV-1: 1.6%; HSV-2: 1.0%, HSV non-typed: 0.5%). Cases due to CMV, EBV, HHV-6, MV, TOSV, HAdV, and LCMV were also detected. For meningoencephalitis, 40.7% of cases were diagnosed, HSV-1 (43.2%) and VZV (27.0%) being the most frequent agents, while cases associated with HSV-2, EV, CMV, MV, and LCMV were also detected. For encephalitis, 27.6% of cases were caused by HSV-1 (71.4%), VZV (19.1%), or EV (9.5%). Other positive neurological syndromes included cerebellitis (EV and HAdV), seizures (HSV), demyelinating disease (HSV-1 and HHV-6), myelopathy (VZV), and polyradiculoneuritis (HSV). No rabies or WNV cases were identified. EVs are the most frequent cause of meningitis, as is HSV for meningoencephalitis and encephalitis. A significant number of cases (42.9% meningitis, 59.3% meningoencephalitis, 72.4% encephalitis) still have no etiological diagnosis.

Molecular epidemiology of enterovirus 71, coxsackievirus A16 and A6 associated with hand, foot and mouth disease in Spain.

Hand, foot and mouth disease (HFMD) is a childhood illness frequently caused by genotypes belonging to the enterovirus A species, including coxsackievirus (CV)-A16 and enterovirus (EV)-71. Between 2010 and 2012, several outbreaks and sporadic cases of HFMD occurred in different regions of Spain. The objective of the present study was to describe the enterovirus epidemiology associated with HFMD in the country. A total of 80 patients with HFMD or atypical rash were included. Detection and typing of the enteroviruses were performed directly in clinical samples using molecular methods. Enteroviruses were detected in 53 of the patients (66%). CV-A6 was the most frequent genotype, followed by CV-A16 and EV-71, but other minority types were also identified. Interestingly, during almost all of 2010, CV-A16 was the only causative agent of HFMD but by the end of the year and during 2011, CV-A6 became predominant, while CV-A16 was not detected. In 2012, however, both CV-A6 and CV-A16 circulated. EV-71 was associated with HFMD symptoms only in three cases during 2012. All Spanish CV-A6 sequences segregated into one major genetic cluster together with other European and Asian strains isolated between 2008 and 2011, most forming a particular clade. Spanish EV-71 strains belonged to subgenogroup C2, as did most of the European sequences circulated. In conclusion, the recent increase of HFMD cases in Spain and other European countries has been due to a larger incidence of circulating species A enteroviruses, mainly CV-A6 and CV-A16, and the emergence of new genetic variants of these viruses.

A novel real-time genotyping assay for detection of the E6-350G HPV 16 variant.

It has been suggested that some E6 human papillomavirus (HPV) type 16 variants could be involved in viral persistence and progression of HPV infection. A novel one-step allelic discrimination real-time PCR was evaluated for E6-350G variant detection in 102 endocervical HPV 16 positive samples. This assay was also used to assess the distribution of this variant in Spanish women with cervical cancer related to HPV 16. The detection limit for the allelic discrimination assay was 50 copies per reaction, even where the E6-350G variant represents only 20% of the variants in the sample. Complete concordance was observed between DNA sequencing and the novel AD RT-PCR assay. Fourteen E6-350T reference strains and 18 E6-350G variants were detected out of 32 endocervical samples from women with cervical cancer. The average age of women who were infected by the E6-350G HPV 16 variant was 10 years lower in these samples than in women who were infected by the reference strain. This novel allelic discrimination assay is a fast, sensitive and specific method for detection of the E6-350G HPV 16 variant.

[Nosocomial bacteremia in the adult patient. Study of associated costs]. / Bacteriemia nosocomial en el paciente adulto. Estudio de costes asociados.

OBJECTIVE: Nosocomial infection causes a prolonged hospital stay and an increase in care costs. The objective of this study was to determine the length of stay excess and costs attributable to nosocomial bacteremia. PATIENTS AND METHODS: Retrospective study of clinical records of 148 patients with nosocomial bacteremia during 1996. A matched case-control study was performed. For matching, the following parameters were used: RDG, year of admission, age 10 years, main diagnosis and number of secondary diagnoses. Costs were determined by excess length of hospital stay and calculating alternative costs. RESULTS: Matching was obtained for 100 cases (67.5%) and cost estimation was performed. Compared with cases, non-matched cases showed differences regarding significant issues for cost, such as hospital stay ( p = 0.01), number of empirical (p = 0.001) or definitive antibiotics (p = 0.03). The median hospital stay for cases was longer than for controls (35 vs 15.5 days, respectively; p = 0.000). When only survivor case-control pairs were considered (n = 75), cases remained in hospital for a median of 36 vs 15 days for controls (p = 0.000). Hospital stay days attributable to nosocomial bacteremia were 19.5 for all matched and 21 for matched survivor cases. Only 76% of cases had stay days attributable to bacteremia. Significant differences between cases and controls included: the mean total costs of admission (p = 0.000), cost of stay (p = 0.001), pharmaceutical expenses (p = 0.000), and cost of microbiological studies (p = 0.000), laboratory work-up (p = 0.001) and radiological studies (p = 0.000). Hospital stay represented more than 60% of costs, followed by pharmaceutical expenses. Cost differences between bacteremic patients and controls, calculated in function of stay median, was 4.424 euros (p = 0.000) and 4.744 euros (p = 0.000) for alternative costs. Ten cases showed a difference that represented more than half of the total difference. CONCLUSIONS: Nosocomial bacteremia represent a stay prolongation and a significant economical burden. Hospital stay and pharmaceutical expenses accounted for the most part of the associated costs. The differences in costs obtained with both methods were small. Since not all selected cases were matched, there may be an error in the appreciation of the difference between cases and controls.

Bacteriemia nosocomial en el paciente adulto. Estudio de costes asociados / Nosocomial bacteremia in the adult patient. Study of associated costs

Fundamento. La infección nosocomial produce una prolongación de la estancia hospitalaria y un incremento del coste de atención. Se pretende determinar el exceso de estancia y los costes atribuibles a la bacteriemia nosocomial. Pacientes y métodos. Estudio retrospectivo de las historias clínicas de 148 pacientes, con bacteriemia nosocomial durante 1996. Se realizó un estudio de casos y controles apareados. Para el apareamiento se utilizaron las siguientes variables: grupos relacionados con el diagnóstico (GRD), año de ingreso, edad ñ 10 años, diagnóstico principal y número de diagnósticos secundarios. Se determinaron los costes mediante prolongación de la estancia y cálculo de costes alternativos. Resultados. Se logró el apareamiento en 100 casos (67,5 por ciento) sobre los que se realizó la estimación de costes. Los casos no apareados presentaron diferencias con los apareados en diversos aspectos significativos tanto para el coste como para la estancia (p = 0,01), número de antibióticos empíricos (p = 0,001) o definitivos (p = 0,03). La mediana de la estancia en los casos fue superior a la de los controles (35 frente a 15,5 días, respectivamente; p = 0,000). Cuando se consideraron solamente los pares con caso y control supervivientes (n = 75), los casos permanecieron en el hospital una mediana de 36 días frente a 15 días los controles (p = 0,000). Los días de estancia atribuible a la bacteriemia nosocomial fueron 19,5 días para todos los pares y 21 días para los pares con caso y control superviviente. Solamente el 76 por ciento de los casos presentó días de estancia atribuibles a la bacteriemia. La diferencia de los costes medios fue significativa entre casos y controles en el coste medio total del ingreso (p = 0,000), coste de la estancia (p = 0,001), coste farmacéutico (p = 0,000) y coste de estudios microbiológicos (p = 0,000), análisis clínicos (p = 0,001) y radiología (p = 0,000).La estancia supuso más del 60 por ciento del coste, seguida del coste farmacéutico. La diferencia del mismo entre los pacientes con bacteriemia y los controles, calculado en función de la mediana de estancia, fue de 4.424 euros (p = 0,000) y de 4.744 euros (p = 0,000) por costes alternativos. Diez casos mostraron una diferencia que supuso más de la mitad de la diferencia total. Conclusiones. La bacteriemia nosocomial supone una prolongación de estancia y una carga económica significativa. La estancia hospitalaria y el coste farmacéutico justificaron la mayor parte del coste asociado. La diferencia de coste obtenida con ambos métodos fue de escasa significación. Al no conseguir el apareamiento de todos los casos seleccionados puede existir un error en la apreciación de la diferencia de costes entre casos y controles (AU)