title: [Investigation of seven different RNA viruses associated with gastroenteritis in children under five years old].
abstract: Viruses are the most frequently detected etiologic agents of gastroenteritis seen in small children. In addition to classical gastroenteritis viruses namely rotavirus, norovirus, adenovirus type 40/41, astrovirus and sapovirus, some novel picornaviruses (Aichi virus, parechovirus, enterovirus) that have been identified in parallel to the developments in molecular diagnostic methods, thought to be associated with diarrhea in humans. However, the data are not enough to prove their actual roles in the pathogenesis of gastroenteritis. The aim of this study was to investigate the presence of rotavirus, norovirus, sapovirus, astrovirus, Aichi virus, parechovirus and enterovirus in the stool samples of children with diarrhoea by reverse-transcriptase polymerase chain reaction (RT-PCR). A total of 50 samples from children admitted to our hospital with diarrhoea between June-December 2012 were included in the study. All the patients were under 5 years of age. Routine bacteriological and parasitological examinations of the patients' stool samples were negative. Total RNAs were extracted from each of the samples and cDNAs were obtained by reverse transcription. All cDNAs were investigated first with the internal control (IC) using PCR. Thirty-one of the 50 cDNAs (62%) were found IC positive. Those 31 samples were further investigated in terms of rotavirus group A and C, norovirus (NoV) genogroup GI and GII, sapovirus, astrovirus, Aichi virus, parechovirus and enterovirus by PCR using specific primer pairs. The predicted sized PCR products obtained were cloned into the pBSK cloning vector and were sequenced. Sequences obtained were subjected to a BLAST search with registered sequences in the GenBank database for the confirmation of the PCR product. Out of 31 RNA positive stool specimens, 12 (38.7%) were found positive for five types of the target viruses. NoV GII (6/31, 19.3%) were detected as the most prevalent virus, followed by NoV GI (2/31, 6.5%), rotavirus group A (2/31, 6.5%), astrovirus (1/31, 3.2%) and sapovirus (1/31, 3.2%). The results of this study revealed that the most frequently detected agents were noroviruses (8/50, 16%) followed by rotavirus (2/50, 4%), astrovirus (1/50, 2%) and sapovirus (1/50, 2%). It was concluded that RT-PCR performed with the primers used in this study could be applied effectively for the molecular epidemiological analysis of RNA viruses leading to gastroenteritis. Larger scale studies conducted in different areas for longer time periods and with a larger population size will provide data for the epidemiological properties of agents of viral gastroenteritis.
authors.forename: Shamim
authors.initials: S
authors.lastname: Akhter
authors.forename: Buse
authors.initials: B
authors.lastname: Turegun
authors.forename: Mehmet
authors.initials: M
authors.lastname: Kiyan
authors.forename: Devran
authors.initials: D
authors.lastname: Gerceker
authors.forename: Haluk
authors.initials: H
authors.lastname: Guriz
authors.affiliation: Ankara University Faculty of Medicine, Department of Medical Microbiology, Ankara, Turkey. fsahin29@hotmail.com.
authors.forename: Fikret
authors.initials: F
authors.lastname: Sahin
mesh_terms: D002675:Child, Preschool; D018076:DNA, Complementary / Q000737:chemistry; D003967:Diarrhea / Q000821:virology*; D005243:Feces / Q000821:virology*; D005759:Gastroenteritis / Q000453:epidemiology / Q000821:virology*; D006801:Humans; D007223:Infant; D012327:RNA Virus Infections / Q000453:epidemiology / Q000821:virology*; D012328:RNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014421:Turkey / Q000453:epidemiology
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral