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Respiratory Syncytial Virus (RSV) is a common, easily spread virus that almost all children catch at least once by the time they turn 2.
RSV spreads just like a common-cold virus, by the touch or someone who has a cold or through coughing sneezing. Background: The present study was to investigate the incidence of the respiratory syncytial virus infection in children and to characterize the RSV circulating in Khartoum state during 2011-12 winter seasons. RSV infections are also responsible for mortality, with almost nine times the mortality rate of influenza [7]. In Sudan work on RSV is very sparse, little information is available about the Incidence of RSV infection in infant, however pneumonia was the first diseases leading to death in 2004, 2005 and 2006 in infants in Sudan [18]. The present study aimed to investigate the incidence of respiratory syncytial virus (RSV) infection in Khartoum state from Gaffer Ebinauff and omdorman children hospitals, in infants and children under 5 years of age with respiratory tract infections (RTI) during 2011-12 winter seasons. DFA was carried out using specific fluoresce in conjugated monoclonal antibody (ARGENE Respiratory Syncytial Virus DFA kit, Varilhes, France) in accordance with the manufacturer’s instructions, for the detection and identification of RSV antigen in direct respiratory specimen. A total of 224 swabs specimens were tested by RT-PCR, out of these RSV was detected in 44 (19.6%) patients.
The figure show percentage of positive and negative RSV cases detected by direct immunofluorescence assays (DFA) and reverse transcription- polymerase chain reaction (RT-PCR).
The figure show percentage of positive RSV cases in Khartoum hospitals with clinical symptoms (Bronchiolitis, Pneumonia, Asthma and Allergy) determined by reverse transcription- polymerase chain reaction (RT-PCR). The Sudanese nucleotide sequences were compared with other sequences published in Gene Bank. The Sudanese RSV-A strains belonged to the same clade with JX645826 A strain (Belgium) and JX131640 A strain (Saudi Arabia) (Figure 4). We in here, provides the first report on molecular detection and characterization of RSV in Sudan. Two hundred and twenty four of samples were collected from children with RTI and significantly high rate of RSV infection was detected by RT-PCR, where by 44 (19.6%) were reported as positive cases.
A limitation of these latter results is that there was a possibility of missing cases due to no visit to the clinic because of mild symptoms and considerable distances between villages and hospitals. The current study showed that the prevalence of RSV was high in 2012 than in 2011 by the two techniques (DFA and RT-PCR), were 75 (67%) and 23 (20.5%) respectively. Although the bronchiolitis is a prominent sign for clinical diagnosis of patients with RSV infection especially in infants [34].
Our data indicated that RSV subtype A is the predominant subtype circulating in Sudan, since all the analyzed samples were from hospitalized outpatients who came from different areas of the country.
Transversionmutation in G gene was identified in one of the Sudanese strains (Isolate 316). The Sudanese RSV viruses are closer to the previous described strains in Saudi Arabia, and that is mainly due to the short distance between Sudan and Saudi Arabia beside the great number of Sudanese people who travel to and from Saudi Arabia for Hajj, umrah and mostly for work.
AcknowledgementsThe authors gratefully acknowledge the Staff of Khartoum Hospital for allowing us to collect the samples, and Central lab for their help in practical work. Authors’ ContributionsSOK carried out the sample collection, virus isolation, DFA, RT-PCR, and drafted the manuscript.
Ethical ClearanceThe study was approved by the Ethical Review Committee (ERC) of the Ministry of Health Khartoum State, Sudan. Although its symptoms are like the common cold, RSV – respiratory syncytial virus – can be deadly. Crystal and Marshall Diehl want to make sure other parents are aware of RSV and know that they, too, can speak up for their child's health care.
After hand-washing, families should try to reduce exposures by avoiding obviously ill people, Dr. A RSV prophylaxis is available for the highest-risk patients, although it is expensive and requires a monthly injection for the entire season. If you manage this site and have a question about why the site is not available, please contact us directly.
But for babies born at 35 weeks or less, or born with hear or lung problems, RSV can lead to serious lung infection, hospitalization, breathing problems and, in some cases, death.
RSV can also be spread by kissing the baby If you have a cold or a fever, limit contact with the baby. Methodology: Throat swab specimens collected from 224 children less than 5 years old, with respiratory tract infections admitted at Khartoum Hospitals in winter season (2011- 2012), were screened for RSV using direct immunofluorescence assay (DFA) and reverse transcription- polymerase chain reaction (RT-). IntroductionRespiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia among infants and children, particularly during the first year of life [1]. Developing countries had the highest mortality rates, with an estimated 66,000 to 199,000 deaths occurring in children younger than five years of age in 2005 [7].
We previously showed an overall sero-prevalence rate of RSV infection in 24 out of 92 (26%) infants with RTI. The epidemiological data and molecular characterization of detected RSV genotype were analyzed.


Data CollectionDemographic data of the patients were collected using a structured questionnaire, which included the following criteria: Age, gender, and clinical symptoms (pneumonia, bronchiolitis, history of asthma and allergy).
Collection of SpecimensA total of 224 throat swab samples (112 samples in each year)were collected from patients by using sterile nylon swabs (Regular Flocked swab, Cat.
RT-PCR for RSVThe reverse transcription- polymerase chain reaction (RT-PCR) assay targeting G gene was performed using a commercial kit following the manufactures instructions (hRSV-Eph-PCR kit, Ecoli, Slovak Republic). Nested RT-PCRThe Nested RT-PCR was carried out for the RT-PCR positive (RSV) samples including cell culture isolate using primer pairs located on the RSV G gene (Oligomicrogen, Korea). Bioinformatics AnalysisSeven consensus sequences of RSV isolates from children less than 5 years of age were successfully analyzed. Clinical Symptoms Associated RSVThe clinical symptoms associated with RSV in positive patients were described in Table 3 and Figure 2. Nucleotide Sequence and Bioinformatics Analysis of RSV Successful sequencing of HRSV based on G gene was done on 7 samples.
In this study we investigated the incidence of respiratory syncytial virus (RSV) infection and characterized the RSV circulating in Khartoum state, during winter. Similar result were recorded in Egypt in which, RSV was detected in 16.4% of the cases [28]. The high rate of RSV in our result explains the increase in RTI cases during winter seasons in Khartoum state. This result may possibly due to the significant variation in RSV outbreaks from year to year [33].
Parallel results were obtained in our study; in which higher percentage of RSV-infected children were males.
Previous studies in Taiwan and Brazil have also documented that Subtype A usually is the predominant circulating strains compared with subtype B [. Several studies of RSV strains have described many mutations in G gene in different positions [J. This is plausible because limited sequence data is available at present time on the prevalence of RSV in Africa, which are more likely to share the same genotypes. ConclusionIn conclusion, this is the first report on the molecular epidemiology of RSV associated RTI in Sudan. RSV is a highly contagious childhood infection that’s responsible for one out of every 13 visits to a pediatrician and one of every 38 trips to the emergency room, a 2009 study from the University of Rochester School of Medicine and Dentistry found. The virus is transmitted through sneezing or coughing, though you'd have to be very close to someone to catch RSV directly. Muskovitz says that in healthy, older infants, children and adults, the signs are usually similar to typical cold symptoms: cough, runny nose, mild fever, possible wheezing. Eisner has been in private practice in Pediatrics in or near the Houston Heights area since July, 1981. Isolation in cell culture followed by nucleotide sequencing and bioinformatics analysis based on the G gene, were done for the RT- positive RSV samples. RSV is a negative-stranded non-segmented RNA virus in the genus Pneumovirus, subfamily Pneumovirinae of the family Paramyxoviridae [2]. Study AreaThe study was conducted in Khartoum State of Central Sudan, Patients involving children under 5 years of age with respiratory tract infection (RTI) seen at the emergency department in Khartoum Hospitals (Gaffer Ebinauff and omdorman children hospitals), were recruited between the 1st of January to the end of March in 2011 and 2012. Then 30 ?L of the suspension sediment was obtained on a slide for immunofluorescence purposes, finally dried then fixed using chilled acetone for 10 min and stored at -20°C for further work. The RNA was extracted from 100 ?L of specimen and 10 ?L of internal control was added to each sample.
Human epithelial type 2 (HEp-2) was obtained from (Vircell, Granada, Spain) and was grown at 37°C in 5% CO2 in modified Eagle's medium supplemented with 10% fetal bovine serum (FBS). There was no statistically significant association between age and RSV infection (P-value=0.903) (Table 1). In 2012 the RSV was detected in 75 (67%) samples using DFA and 23 (20.5%) samples using RT-PCR. However our study showed that there was no statistically significant difference between males and females in having RSV infection (P-value=0.699) (Table 4). Phylogenetic analysis of nucleotide sequences revealed that all analyzed RSV belonged to the RSV-A strain. The Bronchiolitis was the main clinical diagnosis in RSV cases in age group 2-5 years, it were 11 (25%). However, statistical analysis on clinical features and hospitalization with RSV rates between male and female patients did not reveal any striking differences. It highlights the significance of RSV as a dominant etiologic agent contributing towards morbidity and mortality especially in young children in Sudan, and need for further work on incidence of viral pneumonias and their impact on public health.
Abdlrahim for the manuscript corrections and Dr.HishamAltayb for his work on bioinformatics anaylisis. Premature infants and those with compromised heart, lungs or immune systems are at heightened risk for serious complications.


Many need hospitalization and supportive therapy to recover." RSV is a winter virus in the northern hemisphere, November through April, so now is the time to be vigilant about prevention. The virus can live on surfaces, such as countertops, desks, phones and computer keyboards, so if you touch these items and then rub your eyes, or touch your nose or mouth, you may become infected. In very young infants and growing premature infants, at highest risk in their first two RSV seasons, the signs can be very different.
520CS01, Copan Diagnostics Inc., Murrieta, Calif, USA) in 3 ml of transport media (UTM-RT, Cat. 1 ml of each sample was inoculated in HEp-2 monolayer (60%–70% confluent) cultures and was maintained in modified Eagle's medium supplemented with 2% fetal bovine serum and 1% penicillin-streptomycin.
Then the nucleotides sequences of G genes were searched for sequences similarity using nucleotide BLAST [22]. However, data collected from annual records of Ministry of Health [18] in Sudan showed that, within the ten top listed diseases causing hospitalization in infants, Pneumonia accounted for (28.3-30%) during 2004-2008. This variant with the results reported by RoyaNikfar [19], in which bronchiolitis was the most frequent clinical diagnoses in children younger than 2 years old, (33%). The detection and characterization of RSV infection may help to further our understanding with respect to acute respiratory infections of children. We are grateful to the children and parents that approved to provide samples for this study from Khartoum hospital.
KAE helped in the work of virus isolation, RT-PCR and help in the drafting of the manuscript.
Although their doctor recommended Audrey receive preventive therapy, the Diehl family had to fight for insurance coverage for the costly injections.
Positive specimens for RSV resulted in PCR fragments of 298 bp and the internal control resulted in PCR fragment of 550 pb. Following adsorption, fresh medium was added, and inoculated cells were observed daily for cytopatic effect (syncytia) for up to 7 days. Highly similar sequences were retrieved from NCBI and subjected to multiple sequence alignment using BioEdit software.
RSV infections were detected in 136 (60.7%) of these patients, using DFA technique (Table 1, Figure 1). Furthermore, on comparison with reference A2 strain (GenBank accession number M74568) and others strains from gene bank, transversionmutation was identified in one of the Sudanese viruses (Isolate 316) at position 220 were C was replaced by A (Figure 3).
Lane NC; negative control, Lane 1, 2, 6 and 7 showing typical band size of (326 pb) corresponding to the molecular size of G genes, 3, 4, 5 and 8 negative samples. And pneumonia was the most frequent clinical diagnoses in children from 2-5 years old, (34%).
These infants frequently feed poorly, are more lethargic, need stimulation to wake up and can become dehydrated. The clinical symptoms including Bronchiolitis, Pneumonia, Asthma and Allergy showed significantly different rates (pConclusions: This is the first report on molecular characterization that describes the circulation of RSV genotype in Sudan.
Virus was harvested when the monolayer demonstrated approximately 75% CPE (usually on days 3 or 4 post- inoculation). Phylogenetic tree of G genes and their evolutionary relationship with those obtained from database was reconstructed using the maximum likelihood online by phylogeny.fr [23]. The reports showed that pneumonia was the first disease leading to death during 2004, 2005 and 2006 accounting for 1126, 918, 814 deaths respectively. The variation in our result may be due to the fact that, acute bronchiolitis children are at risk for developing pneumonia. DFA and RT-PCR offers rapid methods for detection of RSV in hospitalized children with Respiratory tract infection (RTI). Samples collected were transported on ice at the same day of collection to the Department of Virology, Central Laboratory (Ministry of Science and Technology) and stored at ?80°C until processed.
The cells were subjected to three successive freeze-thaw cycles followed by re-suspension in fresh medium. The Nested RT-PCR products were purified and sequenced by Macrogen’s sequencing service (Amsterdam, Netherlands).
The seven sequences have been deposited with the Gene Bank under accession numbers (AB979187, AB979188, AB979189, AB979190, AB979191, AB979192 and AB979193). A recent study in Khartoum state reported that, the RSV infection was detected in 27 out of 334 patients that were negative for influenza viruses [27].



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