Viral Respiratory Infections with Bocavirus in Romania

Allander et al. at the Karolinska Institutet in Stockhlom, Sweden, first cloned this new member of the family of Parvoviridae in 2005 from pooled nasopharyngeal aspirates (NPA, collection of aspirated fluid from the back of the nasal cavity) [1]. They used a novel technique called molecular virus screening, based on random cloning and bioinformatical analysis. This technique has led to the discovery of new viruses such as polyomavirus KI (Karolinska Institute) [2] and WU (Washington University) [3], which are closely related to each other and have been isolated from respiratory secretions.


Introduction
Allander et al. at the Karolinska Institutet in Stockhlom, Sweden, first cloned this new member of the family of Parvoviridae in 2005 from pooled nasopharyngeal aspirates (NPA, collection of aspirated fluid from the back of the nasal cavity) [1]. They used a novel technique called molecular virus screening, based on random cloning and bioinformatical analysis. This technique has led to the discovery of new viruses such as polyomavirus KI (Karolinska Institute) [2] and WU (Washington University) [3], which are closely related to each other and have been isolated from respiratory secretions.
The name BocaVirus is derived from Bovine and CAnine, referring to the known hosts for other members of this genus; the bovine parvovirus which infects cattle and the minute virus of canines which infects dogs [4].
Parvoviruses (Latin: small viruses) have a 5 kilobase long single stranded DNA.
Full spectrum of diseases associated with human BoV remains to be defined. Presence of the virus in respiratory secretions, serum, and stool suggests that may cause systemic diseases.
The aim of the study was to find that BoV is also present in Romania.

Materials and Methods
The study was part of the SARI (Severe Acute Respiratory Infections) surveillance system which has been set up in Romania since 2009. During the season 2011-2012, the surveillance system comprised 26 hospitals (in Bucharest and 8 counties). The case definition of SARI [5] for the patient's ≥ 5 years old (with onset during the previous 7 days before hospitalization) consists of: fever >38°C, AND cough or sore throat, AND shortness of breath or difficulty breathing.
Case definition of SARI in children <5 years old is the definition for pneumonia and severe pneumonia [5] (

Institute of Research and Development Microbiology and Immunology
Cantacuzino that performed the laboratory confirmation.
We conducted the study between September 2011 and September 2012 to find that BoV is also present in Romania or not.
The samples analyzed were nasal swabs from 309 patients aged 19 days-96 years which were admitted to hospitals from Bucharest and 8 counties from Romania. The samples were transported to the laboratory in viral transport medium, on the same day and processed either on the same day or stored at minus 80°C and processed subsequently.

Results and Discussion
In Romania, the influenza surveillance is performed by the Romanian National Public Health Institute (NPHI) in Bucharest and the National Influenza Centre (NIC) in "Cantacuzino" NIRDMI. There is a network of sentinel general practitioners (GPs) from each county (total number of GPs=313) who weekly report to NPHI the number of patients who consult them with an influenza-like illness (ILI) and a network of severe acute respiratory infections (SARI) (total number of units care=26  (Table 2 and Table 3).

Severe Pneumonia Very Severe Pneumonia Bronchiolitis of infancy
Children below 2 months -symptoms: cough or difficult breathing and -signs: 60 or more breaths per minute, or severe chest indrawing -and no general danger signs (the sign 'stopped feeding well' in young infants replaces 'unable to drink' of the older children as a danger sign), wheezing, stridor in calm child or fever or low body temperature -symptoms: cough or difficult breathing -and signs: general danger signs, wheezing, stridor in a calm child or fever or low body temperature -Clinically diagnosed respiratory condition presenting with breathing difficulties, cough, poor feeding, and irritability and, in the very young, apnea. These clinical features, together with wheeze and/or crepitations on auscultation combine to make the diagnosis.
-bronchiolitis most commonly presents in infants aged three to six months    Of the 309 samples tested, 10 were positive for humanBoV ( Figure 1). All samples were collected from children (7 months-3.5 years) with symptoms of bronchiolitis [4] and pneumonia [6] without gastrointestinal manifestations.The children with pneumonia had interstitial infiltrates noted by chest radiography. When characteristics of the children positive for BoV were compared with those of children with other demonstrated virus infections, no clinically significant differences were recorded.While infections with RSV peaked during the first six months of age, most BoV infections occured ot the age of 1-3 years.
Infections with BoV were found year-round, though most occured in the winter months.
The percent of 3.3% found in our study is lower than the percent found in other countries 9% in Jordan [14], 5.6% in Taiwan [15]. When comparing the detection frequencies, this difference in detection methods has to be taken into consideration. In general PCR assays are more sensitive than antigen detection methods [16]. Therefore it is likely that the true prevalence of the respiratory viruses that were analyzed by IFA is actually higher than here reported. Another potential reason for differing infection frequencies between studies may be due to regional and temporal differences in the incidence of BoV infection.

Conclusions
The human BocaVirus is also present in Romania, meeting at young ages children.
The percent of 3. 3% is similar to that found in Sweden, but lower than in other countries.
The symptoms of patients hospitalized and diagnosed with BocaVirus were only respiratory and no digestive.
Human Bocavirus positive cases were not associated with other viruses.