Występowanie oraz charakterystyka molekularna oporności na azole wśród klinicznych i środowiskowych izolatów Aspergillus fumigatus - Publication - Bridge of Knowledge

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Występowanie oraz charakterystyka molekularna oporności na azole wśród klinicznych i środowiskowych izolatów Aspergillus fumigatus

Abstract

Celem badań prowadzonych w ramach niniejszej pracy doktorskiej było sprawdzenie występowania oporności na azole grzybów A. fumigatus pochodzących z Polski oraz jej molekularna charakterystyka. W toku badań utworzono kolekcję 200 izolatów A. fumigatus pochodzących z różnych miejsc Polski. W skład kolekcji wchodziły izolaty kliniczne, środowiskowe i weterynaryjne. Przynależność gatunkową badanych grzybów potwierdzono metodami diagnostyki klasycznej oraz molekularnej. Następnie oznaczono lekowrażliwość badanych izolatów na posakonazol itrakonazol i worikonazol. W toku przeprowadzonych badań wykazano obecność izolatów opornych na azole. W ramach pracy nad niniejszą rozprawą przeanalizowano występowanie różnych mechanizmów oporności na azole u badanych izolatów: mutacje punktowe w genie CYP51A, występowanie tandemowych powtórzeń w rejonie promotora genu CYP51A (TR34), ekspresję genów CYP51A, CYP51B, MDR1, MDR2, MDR3, ATRF. W toku przeprowadzonych badań zaobserwowano występowanie różnych mutacji w genie CYP51A w tym jednej, którą udało się powiązać z opornością na azole (mutacja L98H skorelowana z TR34), a także zaobserwowano różnice w poziomach ekspresji badanych genów pomiędzy izolatami opornymi i wrażliwymi na azole. W trakcie badań własnych sprawdzono także wpływ obecności azoli w medium hodowlanym na grzyby z gatunku A. fumigatus. Sprawdzono wpływ obecności worikonazolu, posakonazolu i itrakonazolu w medium hodowlanym na poziomy ekspresji genów i zmiany wartości MIC. W toku przeprowadzonych badań zaobserwowano nabycie oporności na azole w wyniku długotrwałej ekspozycji, zmiany poziomów ekspresji badanych genów oraz zmiany w morfologii grzybów w obecności posakonazolu i itrakonazolu. W toku przeprowadzonych badań oznaczono także wrażliwość izolatów na izawukonazol - nowy związek azolowy dopuszczony do stosowania u ludzi. Zaobserwowano występowanie oporności na izawukonazol zarówno izolatów opornych jak i wrażliwych na pozostałe azole.

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Category:
Thesis, nostrification
Type:
praca doktorska pracowników zatrudnionych w PG oraz studentów studium doktoranckiego
Language:
Polish
Publication year:
2018
Bibliography: test
  1. W Polsce występują zarówno kliniczne, środowiskowe i weterynaryjne izolaty A. fumigatus oporne na azole. Wśród przebadanych 200 izolatów oporność wynosiła 8,5% (w tym wśród klinicznych 3,6%, wśród środowiskowych 2,2% wśród weterynaryjnych 78,6%).
  2. Wśród izolatów A. fumigatus występujących w Polsce istnieją różne mechanizmy oporności na azole. Należą do nich mutacja TR 34 L98H w genie CYP51A oraz nadekspresja genów kodujących pompy błonowe i genów odpowiedzialnych za poprawną budowę ściany i błony komórkowej grzybów. open in new tab
  3. Obecność w medium hodowlanym posakonazolu lub itrakonazolu może powodować zmiany w kolorze grzybni grzybów z gatunku A. fumigatus. open in new tab
  4. Obecność w medium hodowlanym badanych azoli indukuje u grzybów z gatunku A. fumigatus zwiększoną produkcję zarodników. open in new tab
  5. Długotrwała obecność itrakonazolu, posakonazolu albo worikonazolu w środowisku może indukować oporność na azole wśród szczepów A. fumigatus. open in new tab
  6. Obecność w medium hodowlanym itrakonazolu i worikonazolu powoduje wzrost ekspresji genów MDR1, MDR2, MDR3, ATRF, CYP51A, CYP51B grzybów A. fumigatus bez względu na mutacje występujące w genie CYP51A. open in new tab
  7. Obecność w medium hodowlanym posakonazolu powoduje wzrost poziomu ekspresji genów MDR1, MDR2, MDR3, ATRF, CYP51A, CYP51B izolatów A. fumigatus bez mutacji TR 34 L98H, zaś w przypadku izolatów z tą mutacją powoduje spadek poziomu ekspresji genów CYP51B, MDR2, MDR3, ATRF a także nie powoduje istotnych zmian w poziomach ekspresji genów CYP51A i MDR1. open in new tab
  8. Nadekspresja genów kodujących transportery błonowe oraz genów odpowiedzialnych za poprawną budowę ściany i błony komórkowej grzybów z gatunku A. fumigatus może przyczynić się do nabycia oporności na azole. open in new tab
  9. Izolaty A. fumigatus oporne wyłącznie na itrakonazol charakteryzują się nadekspresją genu CYP51A.
  10. Izolaty oporne na worikonazol, itrakonazol i posakonazol nie są wrażliwe na działanie izawukonazolu, a inne izolaty A. fumigatus mogą być oporne na izawukonazol pomimo braku oporności na pozostałe triazole. BIBLIOGRAFIA 1. open in new tab
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  215.  Kurzyk E.: "Budowa biofilmu chorobotwórczych drożdży i jego wpływ na oporność." Dokonania Młodych Naukowców nr 5 s. 266-267, 2014
  216. Prezentacje ustne na konferencjach:  Kurzyk E., "Oporność izolatów Aspergillus fumigatus na izawukonazol" Ogólnopolska Konferencja Biotechnologów i Mikrobiologów BioMillenium 2017, Gdańsk 6-8 wrzesień 2017.
  217.  Kurzyk E.: "Quorum sensing -rola w patogenezie drożdży Candida. Wpływ Młodych Naukowców na osiągnięcia Polskiej Nauk, VI Edycja, Gdańsk 25.04.2014
  218. Plakaty na konferencjach:  Nawrot U., Wieliczko A., Kurzyk E., Brillowska-Dąbrowska A.: "Low frequency of itraconazole resistance fund among avian Aspergillus fumigatus isolates" 8 th Trends in Medical Mycology, Mycoses. Serbia, 06-09.10.2017.
  219.  Kurzyk E., Mroczyńska M., Nawrot U., Włodarczyk K., Brillowska-Dąbrowska A.: " Oporność izolatów Aspergillus fumigatus na azole" XXVII Zjazd Polskiego Towarzystwa Mikrobiologów. Bydgoszcz 25-27.09.2016
  220.  Kurzyk E., Nawrot U., Włodarczyk K., Brillowska-Dąbrowska A.: "Występowanie tandemowych powtórzeń (TR 34 ) w rejonie promotora genu CYP51A oraz mutacji w tym genie jako czynników predysponujących do oporności na azole wśród izolatów Aspergillus fumigatus" II Ogólnopolska Konferencja "Drobnoustroje w świecie człowieka -Drobnoustroje oportunistyczne" Bydgoszcz 21-22.05.2016. -praca wyróżniona I miejscem w sesji plakatowej  Mroczyńska M., Kurzyk E., Brillowska-Dąbrowska A., Gucwa K., Szweda P.: Genotypowanie klinicznych izolatów Candida albicans metodą MP PCR" II Ogólnopolska Konferencja "Drobnoustroje w świecie człowieka -Drobnoustroje oportunistyczne" Bydgoszcz 21-22.05.2016.
  221.  Kurzyk E., Mroczyńska M., Nawrot U., Włodarczyk K., Brillowska-Dąbrowska A.: "Czy istnieje konieczność oznaczania oporności na azole dla szczepów Aspergillus fumigatus izolowanych w Polsce?" BioMedSession, Gdańsk 12.12.2015
  222.  Kurzyk E., Nawrot U., Mroczyńska M., Włodarczyk K., Ussowicz M., Zdziarski P., Arendrup MC., Brillowska-Dąbrowska A.: Detection of clinical Aspergillus fumigatus isolates resistant to triazoles. 7th Trends in Medical Mycology. Vol. 58 Supplement S4. P006. str. 53-54. Lizbona 9-12 październik 2015
  223.  Nawrot U., Kurzyk E., Włodarczyk K., Ussowicz M., Arendrup MC., Brillowska-Dąbrowska A.: The first Polish report of azole resistant clinical Aspergillus fumigatus isolate. 6th International Weigl Conference on Microbiology. Acta Biochemica Polonica. Vol. 62 Supplement 2/2015. V.P.38. str. 104. Gdańsk, 8-10 lipiec 2015
  224.  Kurzyk E., Kadzewicz K., Romanowska E., Dzierżanowska-Fangrat K., Brillowska-Dąbrowska A., Szweda P.: "Genotypowanie klinicznych izolatów Candida krusi. " I Ogólnopolska Konferencja "Drobnoustroje oportunistyczne" Bydgoszcz 18-20 wrzesień 2014
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  • Praca współfinansowana ze środków NCN w ramach projektu OPUS nr 2013/11/B/NZ7/04935:
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