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Ugotavljanje pojavnosti virusa Sindbis v Sloveniji
ID Pozvek, Patricija (Author), ID Korva, Miša (Mentor) More about this mentor... This link opens in a new window

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Abstract
Arbovirusi so virusi, ki v naravi krožijo med členonožnimi prenašalci, komarji, klopi in peščenimi muhami, in naravnimi gostitelji kot so ptice in mali sesalci. Ljudje se okužijo z arbovirusi ob vbodu okuženega prenašalca, kadar je gostota prenašalcev in gostiteljev v okolju zelo visoka. Med medicinsko pomembne arboviruse sodi tudi virus Sindbis (SINV), ki pri ljudeh povzroča mrzlico z izpuščaji in bolečinami v sklepih. SINV so v komarjih in pticah dokazali v številnih evropskih državah, bolezen pri ljudeh pa je pogostejša na Finskem in Švedskem. Z magistrsko nalogo smo želeli ugotoviti ali SINV kroži tudi med komarji v Sloveniji, katere vrste komarjev so vpletene v enzoonotski krog virusa in kateri genotip SINV prenašajo. Pregledali smo 97.859 komarjev, ki smo jih vzorčili med leti 2017 in 2023 na 226 lokacijah po vsej Sloveniji. Ujete komarje smo morfološko identificirali do vrste, ter združili v 10.997 skupin glede na lokacijo, datum vzorčenja, vrsto in spol. Iz vsake skupine komarjev smo osamili NK in z molekularnimi metodami ugotavljali prisotnost virusne RNA. RNA SINV smo dokazali v dveh skupinah komarjev vrste Culex modestus, ki smo jih ujeli avgusta 2022 ob Ledavskem jezeru na Goričkem. Ugotovili smo, da je minimalna stopnja okužbe v komarjih znašala 0,09. Glede na nukleotidno zaporedje gena za nestrukturno beljakovino 4, se SINV-SLO uvršča v genotip IV, ki so ga opisali v Rusiji, Azerbajdžanu in na Kitajskem. Na podlagi poravnav med različnimi izolati SINV-IV smo načrtovali amplikone v velikosti 400 bp, s katerimi smo uspešno pomnožili celoten genom SINV-SLO in tako prvič opredelili SINV v Sloveniji.

Language:Slovenian
Keywords:arbovirusi, virus Sindbis, komarji, genotip IV, NGS, filogenetska sorodnost
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[P. Pozvek]
Year:2024
PID:20.500.12556/RUL-160256 This link opens in a new window
UDC:578.833:578.2
COBISS.SI-ID:205758211 This link opens in a new window
Publication date in RUL:24.08.2024
Views:255
Downloads:79
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Secondary language

Language:English
Title:Investigation of the occurrence of Sindbis virus in Slovenia
Abstract:
Arboviruses are viruses that circulate in nature between arthropod vectors, mosquitoes, ticks and sandflies, and natural hosts such as birds and small mammals. Humans become infected with arboviruses through the bite of an infected vector when the density of vectors and hosts in the environment is very high. Medically important arboviruses include the Sindbis virus (SINV), which causes fever, rashes and joint pain in humans. SINV has been demonstrated in mosquitoes and birds in many European countries, but the disease in humans is more common in Finland and Sweden. With the master's thesis, we wanted to find out whether SINV also circulates among mosquitoes in Slovenia, which types of mosquitoes are involved in the enzoonotic cycle of the virus and which genotype of SINV they transmit. We examined 97,859 mosquitoes that were sampled between 2017 and 2023 at 226 locations throughout Slovenia. Mosquitoes were morphologically identified to species and grouped into 10,997 groups based on location, sampling date, species and sex. We isolated RNA from each group of mosquitoes and determined the presence of viral RNA using molecular methods. SINV RNA was detected in two groups of Culex modestus mosquitoes, which we caught in August 2022 by the lake Ledava in Goričko. We calculated that the minimum level of infection in mosquitoes was 0.09. Based on the nucleotide sequence of the nonstructural protein 4 gene, SINV-SLO is classified as genotype IV, which has been described in Russia, Azerbaijan, and China. Based on alignments between different SINV-IV isolates, we designed 400 bp amplicons with which we successfully amplified the entire SINV-SLO genome and thus identified SINV in Slovenia for the first time.

Keywords:arboviruses, Sindbis virus, mosquitoes, genotype IV, NGS, phylogenetic analysis

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