Epidemiological and molecular investigation of a rubella outbreak, Romania, 2011 to 2012.

We describe a rubella outbreak that occurred in Romania between September 2011 and December 2012. During this period 24,627 rubella cases, 41.1% (n=10,134) of which female, were notified based on clinical criteria, and a total of 6,182 individuals were found serologically positive for IgM-specific rubella antibody. The median age of notified cases was 18 years (range: <1-65) and the most affected age group 15 to 19 years (n=16,245 cases). Of all notified cases, 24,067 cases (97.7%) reported no history of vaccination. Phylogenetic analysis of 19 sequences (739 nucleotides each), from 10 districts of the country revealed that the outbreak was caused by two distinct rubella virus strains of genotype 2B, which co-circulated with both temporal and geographical overlap. In addition to the 6,182 IgM-positive rubella cases, 28 cases of congenital rubella syndrome (CRS) were identified, including 11 neonatal deaths and one stillbirth. The outbreak underscores the need to encourage higher vaccination uptake in the population, particularly in women of reproductive age, and to strengthen epidemiological and laboratory investigations of suspected rubella cases. Genetic characterisation of wild-type rubella virus is an essential component to enhance surveillance and here we report rubella virus sequences from Romania.

Differentiation of influenza B lineages from clinical samples by one-step real-time RT-PCR.

BACKGROUND: Influenza viruses type A and type B are a leading cause of annual epidemics in human populations. Since the 1970s, influenza B viruses have diverged into two antigenically distinct virus lineages called the Yamagata and Victoria lineages. We describe the validation and implementation of a one-step real-time RT-PCR (rRT-PCR) assay that can differentiate between the two genetic lineages of type B. METHODS: Validation of rRT-PCR method was carried out using quantified positive control and reference influenza viruses with specific minor groove binder (MGB) probes. The assay was applied on 102 clinical specimens detected positive for influenza type B. RESULTS: Detection limit was found to be as low as 7.95 RNA copies per reaction. The interassay variability and intra-assay variability were found to be low, and comparable for Yamagata and Victoria lineages. No cross-reactivity with the tested subtypes of influenza type A, known to cause human infections, was noticed. Differentiation of influenza B lineages by rRT-PCR was successfully achieved on all of the known positive type B samples. From the total number of clinical specimens tested, 85 samples belonged to B/Yamagata and 17 samples to B/Victoria lineage. CONCLUSION: Differentiation of genetic lineage B influenza virus circulating in Romania in the next seasons by one-step real-time RT-PCR method will supplement the classical test, haemagglutination inhibition (HI), which requires growing of the virus. This method can be advantageous for a balanced selection of samples, in case of lineages co-circulation, for genetic and antigenic characterization.

Single and multipathogen viral infections in hospitalized children with acute respiratory infections.

We aimed to describe the viral etiology of acute respiratory tract infections in children aged 0-8 years admitted to Grigore Alexandrescu Hospital, the largest pediatric hospital in Romania. The patients had clinical diagnosis of pneumonia, bronchiolitis or viral respiratory infections and had been hospitalized between September 2010 and September 2011. The study was part of the "Molecular investigations of acute respiratory infections caused by non-influenza viruses, to assess the implications of infant and young child pathology" (2008-2011), a National Project II--42-164 (MIRVI). We included in the study 241 children that were swabbed in the first 8 days of the onset with the following symptoms during the previous 7 days: fever > 38 degrees C, AND cough or sore throat, and shortness of breath or difficulty breathing .We identified by RT-PCR 131 (54.4%) positive samples: 112 (85.5%) for a single pathogen, 18 (13.7%) for coinfection with two pathogens and 1(0.8%) for coinfection with three pathogens. The most frequent pathogen identified was respiratory syncytial virus (RSV) (40.18%), followed by Rhinovirus (RhV) (20.54%) and human Metapneumovirus (hMPV) (12.50%). We extrapolated our data to the National program of surveillance of SARI (severe acute respiratory infections). In this program, 191 children aged one month-8 years, were hospitalized in the same period, in which the highest percentage of positivity was due to Influenza viruses (62.65%), but RSV was identified with almost the same percent like in MIRVI (32.53%). It should be noted that among patients with pneumonia, bronchiolitis or respiratory viral infections were identified as the causal agent RhV.

Survival of H5N1 influenza virus in water and its inactivation by chemical methods.

The ability of H5N1 Avian Influenza Virus (AIV) to survive in surface water has been assessed in experimental laboratory conditions, based on non-pathogenic avian reassortant model, by titration of infectivity (TCID50) at different time intervals, in three different types of water. The effect of different chemicals on AIV's survival was assessed using the same type of experimental model. After exposure to the chemical, followed by growth on a suitable substrate, the AIV was quantified by a real-time quantitative reverse transcriptase PCR (qRT-PCR). The reassortant virus persisted, and remained infective in aquatic environments, for 12 days at 22-35 degrees C and up to 20 days at 4 degrees C, irrespective of the type of water, supporting the hypothesis of a potential risk for transmitting the virus among birds and contaminating the household water via common sources of water. A significant decrease for AIV persistence models was recorded for sea water, after 12 days, at 35 degrees C. An effective inactivation has been shown when using commercially available products based on glutaraldehyde and penta potassium bis (peroxy mono sulphate) bis(sulphate), respectively. This rapid and safe method for decontamination, developed in this study, might be helpful in implementation of biosafety measures in laboratory and farms against AIV.

Spread of measles virus D4-Hamburg, Europe, 2008-2011.

A new strain of measles virus, D4-Hamburg, was imported from London to Hamburg in December 2008 and subsequently spread to Bulgaria, where an outbreak of >24,300 cases was observed. We analyzed spread of the virus to demonstrate the importance of addressing hard-to-reach communities within the World Health Organization European Region regarding access to medical care and vaccination campaigns. The D4-Hamburg strain appeared during 2009-2011 in Poland, Ireland, Northern Ireland, Austria, Greece, Romania, Turkey, Macedonia, Serbia, Switzerland, and Belgium and was repeatedly reimported to Germany. The strain was present in Europe for >27 months and led to >25,000 cases in 12 countries. Spread of the virus was prevalently but not exclusively associated with travel by persons in the Roma ethnic group; because this travel extends beyond the borders of any European country, measures to prevent the spread of measles should be implemented by the region as a whole.