Analysis of the performance of recombinant nucleoprotein and mucin-like domain proteins produced from Chlamydomonas reinhardtii in the diagnosis of Crimean-Congo Hemorrhagic Fever Virus

Project no. 30837 · Status: Kapanmış (Yayın bekleniyor) · 2023

Summary

<p>Crimean-Congo&nbsp;hemorrhagic&nbsp;fever&nbsp;(CCHF)&nbsp;is&nbsp;an&nbsp;acute,&nbsp;highly&nbsp;contagious,&nbsp;and&nbsp;life-threatening&nbsp;viral&nbsp;infection&nbsp;with&nbsp;a&nbsp;case&nbsp;fatality&nbsp;rate&nbsp;of&nbsp;up&nbsp;to&nbsp;30%.&nbsp;The&nbsp;causative&nbsp;CCHF&nbsp;virus&nbsp;is&nbsp;an&nbsp;orthonairovirus&nbsp;from&nbsp;the&nbsp;Nairoviridae&nbsp;family.&nbsp;The&nbsp;virus&nbsp;is&nbsp;transmitted&nbsp;to&nbsp;humans&nbsp;through&nbsp;the&nbsp;bite&nbsp;of&nbsp;an&nbsp;infected&nbsp;tick&nbsp;or&nbsp;through&nbsp;direct&nbsp;contact&nbsp;with&nbsp;infected&nbsp;body&nbsp;fluids&nbsp;of&nbsp;patients.&nbsp;The&nbsp;ixodid&nbsp;tick,&nbsp;Hyalomma&nbsp;marginatum&nbsp;marginatum,&nbsp;plays&nbsp;a&nbsp;significant&nbsp;role&nbsp;in&nbsp;the&nbsp;transmission&nbsp;of&nbsp;the&nbsp;disease&nbsp;and&nbsp;is&nbsp;the&nbsp;natural&nbsp;reservoir&nbsp;of&nbsp;the&nbsp;CCHF&nbsp;virus.&nbsp;CCHF&nbsp;is&nbsp;a&nbsp;neglected&nbsp;disease&nbsp;and&nbsp;is&nbsp;re-emerging&nbsp;due&nbsp;to&nbsp;climate&nbsp;change&nbsp;affecting&nbsp;tick&nbsp;distribution&nbsp;worldwide.&nbsp;This&nbsp;is&nbsp;supported&nbsp;by&nbsp;the&nbsp;presence&nbsp;of&nbsp;CCHF&nbsp;in&nbsp;more&nbsp;than&nbsp;30&nbsp;countries&nbsp;in&nbsp;parts&nbsp;of&nbsp;Africa,&nbsp;Asia,&nbsp;Eastern&nbsp;Europe,&nbsp;and&nbsp;the&nbsp;Middle&nbsp;East.&nbsp;Furthermore,&nbsp;there&nbsp;is&nbsp;still&nbsp;a&nbsp;need&nbsp;to&nbsp;develop&nbsp;new&nbsp;diagnostic&nbsp;tests&nbsp;with&nbsp;high&nbsp;sensitivity&nbsp;and&nbsp;specificity&nbsp;for&nbsp;the&nbsp;diagnosis&nbsp;of&nbsp;acute&nbsp;or&nbsp;chronic&nbsp;infection&nbsp;in&nbsp;humans.&nbsp;In&nbsp;this&nbsp;project,&nbsp;compared&nbsp;with&nbsp;the&nbsp;preferred&nbsp;expression&nbsp;systems&nbsp;such&nbsp;as&nbsp;E.&nbsp;coli&nbsp;and&nbsp;S.&nbsp;cerevisiae;&nbsp;The&nbsp;green&nbsp;alga&nbsp;Chlamydomonas&nbsp;reinhardtii&nbsp;was&nbsp;chosen&nbsp;as&nbsp;a&nbsp;protein&nbsp;expression&nbsp;system&nbsp;due&nbsp;to&nbsp;its&nbsp;significant&nbsp;advantages,&nbsp;including&nbsp;possessing&nbsp;cellular&nbsp;organelles&nbsp;capable&nbsp;of&nbsp;performing&nbsp;post-translational&nbsp;modifications&nbsp;crucial&nbsp;for&nbsp;the&nbsp;correct&nbsp;folding&nbsp;of&nbsp;any&nbsp;recombinantly&nbsp;produced&nbsp;protein,&nbsp;exhibiting&nbsp;faster&nbsp;growth&nbsp;compared&nbsp;to&nbsp;mammalian&nbsp;cell&nbsp;systems,&nbsp;and&nbsp;being&nbsp;easy&nbsp;and&nbsp;low-cost&nbsp;to&nbsp;produce.&nbsp;Based&nbsp;on&nbsp;this,&nbsp;the&nbsp;aim&nbsp;was&nbsp;to&nbsp;optimize&nbsp;the&nbsp;recombinant&nbsp;production&nbsp;parameters&nbsp;of&nbsp;CCHF&nbsp;virus&nbsp;nucleoprotein&nbsp;(NP)&nbsp;and&nbsp;mucin-like&nbsp;domain&nbsp;(MLD)&nbsp;proteins&nbsp;in&nbsp;C.&nbsp;reinhardtii&nbsp;and&nbsp;to&nbsp;determine&nbsp;the&nbsp;performance&nbsp;of&nbsp;the&nbsp;optimized&nbsp;recombinant&nbsp;production&nbsp;system&nbsp;in&nbsp;CCHF&nbsp;virus&nbsp;diagnosis&nbsp;using&nbsp;serum&nbsp;samples&nbsp;collected&nbsp;from&nbsp;well-categorized&nbsp;CCHF&nbsp;virus-positive&nbsp;individuals.</p>

View details