Određivanje virulentnosti i molekularna tipizacija izolata Bacillus anthracis

master's thesis

• Title (english): Determination of virulence and molecular typing of Bacillus anthracis isolates
• Author: Ivana Steiner
• Mentor: Jadranka Frece (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for General Microbiology and Food Microbiology); Zagreb
• Defense date and country: 2011-06-06, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Bacillus anthracis je sporogena i virulentna bakterija, patogen koji uzrokuje ozbiljnu i često smrtonosnu bolest bedrenicu (antraks). Antraks je zoonoza koja se može prenijeti na čovjeka i uzrokovati smrtonosnu ili lokalnu infekciju. Bakterija tvori spore koje preživljavaju desetljecima, a mogu se upotrijebiti i kao biološko oružje. B. anthracis, kao infektivni biološki agens, predstavlja veliki rizik i potencijalnu opasnost za čovjeka i određene životinje, stoga postoji velika potreba za brzom i detaljnom identifikacijom i molekularnom tipizacijom izolata za potrebe epidemioloških istraživanja. B. anthracis je izrazito genetički monomorfna vrsta te se tipizacija temelji na analizi varijabilnih regija poput VNTR lokusa. Glavni faktori virulencije B. anthracis, toksini i kapsula, kodirani su na dva plazmida virulencije pXO1 odnosno pXO2. U ovom radu analizirano je 14 sojeva B. anthracis izdvojenih tjekom infekcija ovaca i goveda od kojih je 13 sa područja Hrvatske i jedan iz Mostara (BiH). Ispitali smo prisutnost gena pagA (pXO1) i gena cap-B/C (pXO2) PCR metodom i pokazali da je devet izolata potpuno virulentno (pXO1+, pXO2+), tri su djelomicno virulentni (pXO1-, pXO2+) dok su dva nevirulentni soj Sterne (pXO1+, pXO2-). Provedena je molekularna tipizacija sojeva metodom analize varijabilnog broja uzastopnih ponavljanja na šest lokusa. Određen je genotip za 12 virulentnih sojeva. Utvrđeno je da su na području Hrvatske i Mostara prisutna dva različita genotipa B. anthracis, A1 i B2.

Abstract (english)

Bacillus anthracis is a sporeforming, virulent bacteria, a pathogen and the causative agent of the serious and often lethal disease anthrax. Anthrax is a zoonosis that can be transmitted to humans causing a lethal or local infection. A bacterium produces spores that can exist in the soil for decades, and can be used as a biological weapon. B. anthracis, as an infectious biological agent poses a threat to humans, domesticated animals and wildlife, and necessitates the need for a rapid and highly specific identification and typing assay in epidemiological studies. The genome of Bacillus anthracis is extremely monomorphic, therefore molecular typing is based on analysis of variable regions like VNTR loci. Main virulence factors of B. anthracis, toxins and capsule are encoded on two virulence plasmids pXO1 and pXO2, respectively. In this diploma thesis 14 isolates, isolated from sheep and cattle, were analysed, 13 were from Croatia and one from Mostar (BiH). The presence of pagA (pXO1) and B/C (pXO2) genes was determined using PCR method. Nine of the strains are fully virulent (pXO1+, pXO2+), three are virulent (pXO1-, pXO2+) and two are avirulent vaccine strain, Sterne (pXO1+, pXO2-). Molecular typing system based on variable-number tandem repeat analysis at six loci was used. Genotype was determined for 12 virulent strains. Analysis identified two distinct genotypes in Croatia and Mostar.

• Keywords: Bacillus anthracis; faktori virulencije; molekularna tipizacija; MLVA
• Keywords (english): Bacillus anthracis; virulence factors; molecular typing; MLVA
• Language: croatian
• URN:NBN: urn:nbn:hr:159:919995
• Study programme: Title: Biotechnology; specializations in: Biochemical-Microbiology; Course: Biochemical-Microbiology; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki (diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-30 08:01:52