De Novo Sequencing and Resurrection of a Human Astrovirus-Neutralizing Antibody.

Monoclonal antibody (mAb) therapeutics targeting cancer, autoimmune diseases, inflammatory diseases, and infectious diseases are growing exponentially. Although numerous panels of mAbs targeting infectious disease agents have been developed, their progression into clinically useful mAbs is often hindered by the lack of sequence information and/or loss of hybridoma cells that produce them. Here we combine the power of crystallography and mass spectrometry to determine the amino acid sequence and glycosylation modification of the Fab fragment of a potent human astrovirus-neutralizing mAb.

Norovirus-associated encephalitis in a previously healthy 2-year-old girl.

We describe the case of 2-year-old girl with encephalitis associated with norovirus infection. The viral genome was detected in cerebrospinal fluid and stool by reverse transcription polymerase chain reaction. This is the first time in Spain and the second worldwide that the genome of norovirus from the stool sample and 2 cerebrospinal fluid samples were genetically characterized and matched.

[Microbiological profile of acute viral gastroenteritis attended in a paediatric department of an area with high vaccine cover against rotavirus].

Introduction: Vaccination against rotavirus has led to a significant decline of the disease. The aim of the present work is to evaluate the clinical and epidemiological features of the viral acute gastroenteritis (AGE) in an area with high immunization coverage against rotavirus.

Method: A prospective microbiological evaluation was made of stool culture and Real Time Polymerase Chain Reaction (RT-PCR) to gastroenteric virus and genotyping of rotavirus strains in < 5 year-old with AGE episodes attended by or admitted to our hospital from November-March of 2009-2010 and 2010-2011.

Monitoring of children with acute gastroenteritis in Madrid, Spain, during 2010-2011: rotavirus genotype distribution after the vaccines introduction.

Background And Objectives: A structured surveillance study was conducted on children with diarrhea who were hospitalized in Madrid (Spain) during 2010-2011, in order to describe temporal, geographic, and age-related trends in rotavirus (RV) strains after the introduction of the RV vaccines in our country.

Study Design And Results: A total of 370 children were enrolled, with RV being detected in 117 (31.6%) cases.

Nosocomial rotavirus gastroenteritis in Spain: a multicenter prospective study.

Background: The objective of this study was to assess the incidence of nosocomial rotavirus gastroenteritis among children <2 years of age.

Methods: We conducted a prospective active surveillance for acute gastroenteritis (AGE) in the pediatric wards of 3 representative hospitals in Valencia (Spain) from October 2006 to March 2007, among children between 1 and 23 months of age with acute diarrhea. Children were followed up for 3 days after discharge.

Prevalence and clinical characteristics of norovirus gastroenteritis among hospitalized children in Spain.

Background: The importance of norovirus as a cause of gastroenteritis outbreaks is well documented, but the role of norovirus in sporadic acute severe gastroenteritis is not so well established. The aim of this study was to determine the prevalence and clinical characteristics of norovirus gastroenteritis among hospitalized children.

Methods: A prospective study was conducted in children less than 5 years old, admitted with acute gastroenteritis between January 2005 and January 2008 to the Pediatrics Department of the Universitary Hospital, Albacete, Spain.

[Detection of a rotavirus G9P[8] outbreak causing gastroenteritis in a geriatric nursing home].

Introduction: The aim of this study was to describe an outbreak of gastroenteritis due to rotavirus in a geriatric nursing home.

Method: Stool samples from 8 patients were studied. Antigen detection was carried out by ELISA, and molecular typing was performed by RT-PCR.

[Rotavirus and other viruses causing acute childhood gastroenteritis].

Acute gastroenteritis is one of the most common diseases, affecting children worldwide. Viruses are recognized as a major cause of this disease, particularly in children. Since the Norwalk virus was identified as a cause of gastroenteritis, the number of viral agents associated with diarrheal disease in humans has progressively increased.

European multicenter evaluation of commercial enzyme immunoassays for detecting norovirus antigen in fecal samples.

A total of 2,254 fecal samples were tested in a European multicenter evaluation of commercially available norovirus antigen detection assays. Two commercial enzyme immunoassays, IDEIA Norovirus (Oxoid; Thermo Fisher Scientific, Ely, United Kingdom) and RIDASCREEN Norovirus (R-Biopharm, Darmstadt, Germany), were included in the evaluation, and their performance was compared with the results of reverse transcription-PCR (RT-PCR). Included in the evaluation were samples collected in sporadic cases of gastroenteritis, samples from outbreaks in which two or more samples were collected, well-characterized samples representing genotypes currently cocirculating within Europe, and samples collected from patients with gastroenteritis caused by a pathogen other than norovirus.

Human rotavirus G9 and G3 as major cause of diarrhea in hospitalized children, Spain.

In Spain, diarrhea remains a major cause of illness among infants and young children. To determine the prevalence of rotavirus genotypes and temporal and geographic differences in strain distribution, a structured surveillance study of hospitalized children <5 years of age with diarrhea was initiated in different regions of Spain during 2005. Rotavirus was detected alone in samples from 362 (55.

[Assessment of two methods of antigenic detection by ELISA for the diagnosis of norovirus outbreaks].

Introduction: The aim of this study was to assess two ELISA techniques for the detection of outbreaks of norovirus.

Method: One-hundred and sixty-five fecal samples from patients affected in 30 outbreaks were studied.

Results: On the basis of a specific consensus criterion between techniques for confirming outbreaks, the sensitivity and specificity was respectively 80% and 90% for RIDASCREEN, and 80% and 100% for IDEIA.

Development of a one step real-time RT-PCR method for sensitive detection of human astrovirus.

Human astrovirus (HAstV) has been recognized as the second most common cause of diarrhoea among children under 5 years old. To date, the true incidence of HAstV was underestimated when using enzyme immunoabsorbent assays (EIAs) and conventional reverse transcription (RT)-polymerase chain reaction (PCR) methods. The sensitivity of detection of EIA is insufficient and, although RT-PCR is more sensitive than EIA, the time required is a limitation for astrovirus detection.

Diversity of group A human rotavirus types circulating over a 4-year period in Madrid, Spain.

The incidence and distribution of human rotavirus G types among children under 5 years old with acute gastroenteritis were determined over a 4-year period (1998 to 2002) by using monoclonal antibodies and reverse transcription-PCR methods. Rotavirus was detected in 1,155 (31%) of 3,760 specimens tested. Rotavirus was studied in every month of the 48-month survey period.

Vaccinia virus recombinant expressing an 87-kilodalton polyprotein that is sufficient to form astrovirus-like particles.

Human astrovirus is an important cause of acute gastroenteritis. We have generated, for the first time, a vaccinia virus recombinant expressing the astrovirus 87-kDa structural polyprotein. The results demonstrate that this expression results in the formation of virus-like particles in the absence of other astrovirus proteins and genomic RNA.

Acute viral gastroenteritis: proportion and clinical relevance of multiple infections in Spanish children.

Dual infections associated with acute infectious diarrhoea and its microbiological, epidemiological and clinical findings have been evaluated in patients selected from a comprehensive survey of children under 4 years old, admitted to hospital emergency rooms from October 1996 to November 1997. A total of 820 children (433 males and 387 females) were enrolled. Stools were tested for rotavirus, adenovirus, astrovirus and bacterial enteropathogens.

Astrovirus acute gastroenteritis among children in Madrid, Spain.

Background: Human astroviruses cause infantile gastroenteritis worldwide, but the prevalence of disease varies greatly by setting. Since 1997 we have conducted a survey to determine the causes of diarrhea among Spanish children attending an emergency room in Madrid and to characterize the clinical features of viral-associated gastroenteritis.

Objectives: To define the epidemiologic role of astrovirus-associated gastroenteritis in Spanish children, to review its clinical features and to compare these illnesses with those caused by rotavirus.