<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article  PUBLIC "-//NLM//DTD Journal Publishing DTD v3.0 20080202//EN" "http://dtd.nlm.nih.gov/publishing/3.0/journalpublishing3.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" dtd-version="3.0" xml:lang="en" article-type="research article"><front><journal-meta><journal-id journal-id-type="publisher-id">AJMB</journal-id><journal-title-group><journal-title>American Journal of Molecular Biology</journal-title></journal-title-group><issn pub-type="epub">2161-6620</issn><publisher><publisher-name>Scientific Research Publishing</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.4236/ajmb.2020.104017</article-id><article-id pub-id-type="publisher-id">AJMB-102752</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Biomedical&amp;Life Sciences</subject></subj-group></article-categories><title-group><article-title>
 
 
  The Prevalence of Human Metapneumovirus among Children with Acute Respiratory Tract Infection in North Kordofan State, Sudan
 
</article-title></title-group><contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Huzifa</surname><given-names>Abdalla Abdelrahman Ahmed</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Nassreldeen</surname><given-names>Khalid Abdelrahman</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Mudathir</surname><given-names>Abdallah Adam</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib></contrib-group><aff id="aff2"><addr-line>Faculty of Medical Laboratory Science, University of Al Fashir, Al Fashir, Sudan</addr-line></aff><aff id="aff1"><addr-line>Faculty of Medicine, Sudan International University, Khartoum, Sudan</addr-line></aff><pub-date pub-type="epub"><day>09</day><month>09</month><year>2020</year></pub-date><volume>10</volume><issue>04</issue><fpage>259</fpage><lpage>264</lpage><history><date date-type="received"><day>6,</day>	<month>May</month>	<year>2020</year></date><date date-type="rev-recd"><day>6,</day>	<month>September</month>	<year>2020</year>	</date><date date-type="accepted"><day>9,</day>	<month>September</month>	<year>2020</year></date></history><permissions><copyright-statement>&#169; Copyright  2014 by authors and Scientific Research Publishing Inc. </copyright-statement><copyright-year>2014</copyright-year><license><license-p>This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/</license-p></license></permissions><abstract><p>
 
 
  Objective: To determine the prevalence of human metapneumovirus (hMPV) among children with acute respiratory tract infection and the correlation between (hMPV) and age group. 
  Methods: This prospective, descriptive cross-sectional hospital-based study is carried out from January to May 2019 among children from under five years old who were admitted to El-Obeid pediatric teaching Hospital, North kordofan, Sudan with acute respiratory tract infection. 
  Results: Fifty hospitalized children with acute respiratory tract infection were enrolled in the study. Two of these (4%) were tested positive for (hMPV) in the age group (7 months - 2 years) old. Moreover, the study showed no significant correlation between infection with (hMPV) and age.
 
</p></abstract><kwd-group><kwd>Human Metapneumovirus</kwd><kwd> Respiratory Tract Infection</kwd><kwd> Real Time</kwd><kwd> Sudan</kwd></kwd-group></article-meta></front><body><sec id="s1"><title>1. Introduction</title><p>Human metapneumovirus (hMPV) is a negative-sense single-stranded RNA virus of the family Pneumoviridae [<xref ref-type="bibr" rid="scirp.102752-ref1">1</xref>]. It was isolated for the first time in 2001 in the Netherlands [<xref ref-type="bibr" rid="scirp.102752-ref2">2</xref>]. It is the second most common cause after human respiratory syncytial virus (RSV) of upper and lower respiratory infection in young children [<xref ref-type="bibr" rid="scirp.102752-ref2">2</xref>].</p><p>Since the discovery of human metapneumovirus (hMPV) the virus has been identified worldwide, the major challenges faced by the medical and scientific communities are the understanding of the pathogenesis of hMPV disease and the development of a safe and effective vaccine to protect against infection and disease caused by this newly recognized respiratory virus [<xref ref-type="bibr" rid="scirp.102752-ref2">2</xref>].</p><p>Worldwide, HMPV prevalence in hospital inpatient or community studies, in children or elderly adults, varies widely from as low as 1.7% to as high as 17%, with generally higher prevalence in outpatients compared to inpatients and, also, more in children younger than 5 years compared to older age groups [<xref ref-type="bibr" rid="scirp.102752-ref3">3</xref>].</p><p>Human metapneumovirus is most likely spread from an infected person to others through secretions from coughing and sneezing, close personal contact, such as touching or shaking hands in addition to touching objects or surfaces that have the viruses on them then touching the mouth, nose, or eyes [<xref ref-type="bibr" rid="scirp.102752-ref4">4</xref>].</p><p>Upon entering the body, hMPV infects the cells in your respiratory tract. The infection of these cells by the hMPV leads to the release of local chemicals and hormones that can trigger the body’s immune response. This response leads to the classic symptoms of a “cold”. In some individuals, the disease can spread to the main airways, or bronchi, causing atypical pneumonia [<xref ref-type="bibr" rid="scirp.102752-ref4">4</xref>].</p><p>In term of detection, the efforts cultivating hMPV virus were faced by difficulties due to its poor growth inconventional cell culture and have no characteristic cytopathic effects. Sensitive (RT-PCR) Molecular techniques, in addition to touch-down genomic target sequences, enzyme-linked immunosorbent assay and immunofluorescence serological test are used in detection of the virus from clinical specimen [<xref ref-type="bibr" rid="scirp.102752-ref5">5</xref>].</p><p>Documented information regarding the prevalence of hMPV disease in Sudan is scarce. Hence, the aim of this study was the generation of preliminary information about hMPV infection among children patients attending El-Obeid pediatric teaching hospital with acute respiratory tract infections, where no such study has previously been conducted.</p></sec><sec id="s2"><title>2. Methods</title><sec id="s2_1"><title>2.1. Time and Duration of the Study</title><p>This was a descriptive cross-sectional hospital-based study, carried out from January to May 2019 among children from zero to five years old whom were admitted to El-Obeid teaching Hospitals, North Kordofan State, Sudan. The hospital is a 250-bed tertiary care facility, which serves as a referral Centre for North Kordofan State. The average patient turnover at the hospital is 50 to 150 patients per day.</p></sec><sec id="s2_2"><title>2.2. Study Population</title><p>Fifty children co-patients at the pediatric ward El-Obeid teaching Hospitals during the study period were asked to answer a structured questionnaire consisting of socio-demographic data (age, gender and residence). The informed consent related volunteers were recruited randomly, any child, with acute respiratory tract infection admitted to the pediatric ward has a chance to be selected a once. Fifty children parents have consented for their children to participate in this study with a moderate rate of refusal.</p></sec><sec id="s2_3"><title>2.3. Sample Collection</title><p>Throat swab samples were obtained from patients by inserting sterile nylon swab (Regular Flocked swab, Cat. No. 520CS01, Copan Diagnostics Inc., Murrieta, Calif, USA) and rubbing the tonsils and the posterior wall of the pharynx. Samples collected were transported in an ice pack on the same day of collection to the Virology lab, Central Laboratory and stored at −80˚C until tested.</p></sec><sec id="s2_4"><title>2.4. RNA Extraction</title><p>Total RNA was extracted by using the QIAamp Viral RNA Mini spin according to the protocol of the manufacturer (Qiagen, Germany). Briefly, 140 μl of throat swab sample was added to 560 μl buffer AVL containing carrier RNA, and then incubated at room temperature for 10 minutes. Subsequently, 560 μl of ethanol (96% - 100%) was added to the sample after which 630 μl of the resulting solution was applied to a column. A volume of 500 μl of AW1 and AW2 was added for washing and the nucleic acids were eluted with 60 μl AVE buffer and stored at −80˚C until used.</p></sec><sec id="s2_5"><title>2.5. Real-Time RT-PCR</title><p>One-step RT-PCR was done to detect viral RNA by using a commercial kit following the manufacturer’s instructions (AgPath-ID<sup>TM</sup> One-step RT-OCR Kit Ambion USA) and primer/probe for hMPV.</p><p>The real-time PCR master mix for one reaction was prepared as follows: 12.5 μl of 2&#215; RT-PCR buffer reaction mix (consisting of a proprietary buffer system, MgSO<sub>4</sub>, dNTPs), 2 μl of primer/0.5 probe, 1 μl enzyme mix, 1 μl enhancer, 3 μl of molecular grade water, and 5 μl of total RNA. The final volume was 25 μl for a single reaction. The reaction was performed in Rotor-gene Q real-time PCR (Germany). The thermal cycling conditions were 15 minutes at 45˚C for reverse transcription, 2 minutes at 95˚C for initial denaturation and 45 cycles of 15 seconds at 95˚C for denaturation and 45 seconds at 60˚C for annealing and extension [<xref ref-type="fig" rid="fig1">Figure 1</xref>].</p></sec><sec id="s2_6"><title>2.6. Ethical Review</title><p>The Ethical Review Committee (ERC) of the Ministry of Health North Kordofan State, Sudan approved the study.</p></sec><sec id="s2_7"><title>2.7. Statistical Analysis</title><p>Data were analyzed using the statistical package (SPSS) version 22.</p></sec></sec><sec id="s3"><title>3. Results</title><p>A total of fifty pediatric patients (less than five years old), were enrolled in this study; 17 were male and 33 were females. The prevalence of hMPV was 4%, 2 (50) by Real-time RT-PCR (<xref ref-type="table" rid="table1">Table 1</xref>).</p><table-wrap id="table1" ><label><xref ref-type="table" rid="table1">Table 1</xref></label><caption><title> Frequency of hMPV among acute respiratory infection patients</title></caption><table><tbody><thead><tr><th align="center" valign="middle"  colspan="4"  >RT PCR Test</th></tr></thead><tr><td align="center" valign="middle" >Gender</td><td align="center" valign="middle" >Frequency (%)</td><td align="center" valign="middle" >Positive</td><td align="center" valign="middle" >Negative</td></tr><tr><td align="center" valign="middle" >Male</td><td align="center" valign="middle" >17 (34%)</td><td align="center" valign="middle" >0</td><td align="center" valign="middle" >17</td></tr><tr><td align="center" valign="middle" >Female</td><td align="center" valign="middle" >33 (66%)</td><td align="center" valign="middle" >2</td><td align="center" valign="middle" >31</td></tr><tr><td align="center" valign="middle" >Total</td><td align="center" valign="middle" >50 (100%)</td><td align="center" valign="middle" >2 (4%)</td><td align="center" valign="middle" >48 (96%)</td></tr></tbody></table></table-wrap><p>The males to females ratio was (0:0.1) in patients who are (hMPV) positive.</p><p>Human MPV positive patients restricted among 7 months to 2 years age groups (<xref ref-type="table" rid="table2">Table 2</xref>). Statistically there is no significant association between infection with hMPV and age group.</p></sec><sec id="s4"><title>4. Discussion</title><p>There is a paucity of information regarding the prevalence of hMPV in sub-Saharan Africa despite a scarce studies documented in Sudan, Kenya and South Africa. This is a first study was conducted in El-Obeid pediatric teaching Hospital North Kordofan state, to determine the prevalence of Human metapneumovirus (HMPV) in children complaining of acute respiratory infection, which was considered upon main causes of morbidity and mortality in children, their behavior tends to be seasonal and vary by geographical locations.</p><p>The present study revealed the prevalence of hMPV infection was 4% out of 50 patients, which was lower compared to study done by Attar and his colleagues in Sudan, whom reported the prevalence of hMPV was 26.5% this difference may be due to seasonal distribution of the virus. A high incidence of hMPV infection observed during the winter-spring season [<xref ref-type="bibr" rid="scirp.102752-ref6">6</xref>].</p><p>A similar results obtained by Mona et al., Al-Turab et al., and Ramirez et al.,</p><table-wrap id="table2" ><label><xref ref-type="table" rid="table2">Table 2</xref></label><caption><title> Distribution of hMPV positive patients age groups</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Age group</th><th align="center" valign="middle" >Frequency</th><th align="center" valign="middle" >Positive</th><th align="center" valign="middle" >Negative</th></tr></thead><tr><td align="center" valign="middle" >0 - 6 months</td><td align="center" valign="middle" >2</td><td align="center" valign="middle" >0</td><td align="center" valign="middle" >2</td></tr><tr><td align="center" valign="middle" >7 months - 2 year</td><td align="center" valign="middle" >27</td><td align="center" valign="middle" >2</td><td align="center" valign="middle" >25</td></tr><tr><td align="center" valign="middle" >2 year - 5 years</td><td align="center" valign="middle" >21</td><td align="center" valign="middle" >0</td><td align="center" valign="middle" >21</td></tr></tbody></table></table-wrap><p>in Egypt, Kuwait and Colombia, showed that incidence of (hMPV) was (4%), (5%) and (5.2%) respectively [<xref ref-type="bibr" rid="scirp.102752-ref7">7</xref>] [<xref ref-type="bibr" rid="scirp.102752-ref8">8</xref>] [<xref ref-type="bibr" rid="scirp.102752-ref9">9</xref>].</p><p>Moreover, in Saudi Arabiaa study performed by Alsuheel et al., found that the prevalence of human metapneumovirus was 9.9%, which higher than the prevalence documented by this study [<xref ref-type="bibr" rid="scirp.102752-ref10">10</xref>].</p><p>Furthermore, a study concluded in China, reported 2.0% for the prevalence of hMPV infections among (RTIs)hospitalized children, which is considered lower than the determine by our study, and the age group of the positive hMPV were 1 to 2 years, while in our study was (7 months - 2 years), which is relatively close to our study findings [<xref ref-type="bibr" rid="scirp.102752-ref11">11</xref>].</p></sec><sec id="s5"><title>5. Conclusion</title><p>This study revealed the existence of hMPV among children patients attending El-Obeid teaching Hospitals, Northern Kordofan State, Sudan that was not previously established in this hospital. These findings are useful for future studies since there is a paucity of information regarding hMPV infection in Sudan.</p></sec><sec id="s6"><title>Acknowledgements</title><p>The author wish to thank El-Obeidpediatric Teaching Hospital and Al-Daman Specialist Hospital for their co-operation and acceptance to conduct our study there. We are deeply indebted to the participants in this study.</p></sec><sec id="s7"><title>Conflicts of Interest</title><p>The authors declare no conflicts of interest regarding the publication of this paper.</p></sec><sec id="s8"><title>Cite this paper</title><p>Ahmed, H.A.A., Abdelrahman, N.K. and Adam, M.A. (2020) The Prevalence of Human Metapneumovirus among Children with Acute Respiratory Tract Infection in North Kordofan State, Sudan. 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