Author/Editor		Poljšak-Prijatelj, Mateja; Frelih, Tatjana; Zimšek, Janet; Berce, Ingrid; Barlič-Maganja, Darja
Title		Kalicivirusi - povzročitelji epidemičnega gastroenteritisa pri otrocih v vrtcu - izkušnje v Sloveniji
Type		članek
Source		Zdrav Vestn
Vol. and No.		Letnik 70, št. 11
Publication year		2001
Volume		str. 619-22
Language		slo
Abstract		Background. Human caliciviruses represent a genetically and antigenically diverse group of single stranded RNA viruses associated with acute gastroenteritis in humans. In last two years the number of notified gastroenteric cases in Slovenia isncreasing. Samples were analysed by direct electron microscopy (EM). For increased sensitivity RT-PCR assay was performed. Using molecular methods for the detection of caliciviruses we tried to detect causative agent in outbreaks where no viruses were found by direct EM. Methods. After onset of outbreak an epidemiologic questionnaire was performed, hygienic testing and determination of pathogenic bacteria. Latex agglutination for determination of rota-and adenoviruses 40/41 were performed. Additionally ELISA test for determination of astroviruses was used. Stool samples were examined by direct electron microscopy. RNA was isolated from IS positive samples in which small roundstructured viruses (SRSVs) were observed. RNA was amplified by RT-PCR using specific primers and the obtained products were confirmed in hybridisation test. Results. 20 children from 97 and 2 adults form 20 became ill. Symptoms included diarrhoea, vomiting and fever lastingfor 1 to 3 days. None of patients sought medical help neither were hospitalized. Obtained food samples were not contaminated. From five hygienic smears coagulase positive staphylococci were isolated. All stool samples tested for pathogenic bacteria were negative, as well as for rota-and adenoviruses. Electron microscopy revealed 15 stool samples containing SRSVs. 11 of them were positive for caliciviruses, corresponding to genogroup II (GG 11), as determined by RT-PCR and hybridisation assay. Conclusions. If an outbreak in the community is reported early or at least soon enough caliciviruses can be identified by RTPCR and hybridisation assay and the outbreak could be restricted to limited area.
Summary		Izhodišča. Človeški kalicivirusi, povzročitelji črevesnih okužb, predstavljajo genetsko in antigensko različno skupino virusov s pozitivno usmerjeno enovijačno RNK. Po podatkih Inštituta za varovanje zdravja v Sloveniji v zadnjih dveh letih narašča število prijavljenih epidemij virusnega gastroenteritisa, povzročenega s kalicivirusi. Do sedaj so bili vsi kalicivirusi dokazani z metodami elektronske mikroskopije. Z izboljšanimi molekularnimi diagnostičnimi metodami želimo dopolniti elektronskomikroskopske metode in tako pojasniti epidemije, pri katerih povzročitelja ne odkrijemo. Metode. Po izbruhu epidemije v vrtcu smo opravili epidemiološko poizvedovanje, odredili pregled brisov na snažnost, preiskavo na patogene črevesne bakterije in lateksno aglutinacijo za določitev rotavirusov in adenovirusov 40/41. Uporabili smo tudi encimskoimunsko metodo za dokaz astrovirusov. Iztrebke bolnikov smo pregledali z metodo neposredne elektronske mikroskopije, izveddi verižno reakcijo s polimerazo s poprejž njo reverzno transkripcijo (RT-PCR) s specifičnimi začetnimi oligonukleotidi in produkte RT-PCR potrdili s hibridizacijo. Rezultati. Z analizo podatkov iz vprašalnikov smo ugotovili, da je po izbruhu epidemije v novogoriškem vrtcu zbolelo z gastroenteritisom 20 od 97 otrok in 2 od 12 zaposlenih. Bolezenski znaki z drisko, bruhanjem in povišano telesno temperaturo so trajali 1 do 3 dni. Nihče od bolnikov ni iskal zdravniške pomoči, niti ni bil hospitaliziran. Odvzeti vzorci hrane so bili neoporečni. Iz brisov na snažnost z delovnih površin smo iz petih brisov izolirali koagulaza pozitivni stafilokok. Iztrebki, ki smo jih preiskali na patogene črevesne bakterije, so bili negativni. vseh 39 iztrebkov, ki smo jih preiskali na rota- in adenoviruse, je bilo negativnih. Z elektronskomikroskopskim pregledom IS iztrebkov smo določili male okrogle strukturtrane viruse. (Izvleček skrajšan pri 2000 znakih).
Descriptors		GASTROENTERITIS CALICIVIRUS CHILD DAY CARE CENTERS DISEASE OUTBREAKS CHILD, PRESCHOOL MICROSCOPY, ELECTRON POLYMERASE CHAIN REACTION