Abaphandi baseDatshi badibanisa i-CRISPR kunye ne-bioluminescence kuvavanyo lokuvavanyaizifo ezosulelayo

Iprotheyini esandula ukwenziwa yasebusuku inokukhawulezisa kwaye yenza lula ukuxilongwa kwezifo zentsholongwane, ngokutsho kwabaphandi baseNetherlands.
Uphononongo lwabo, olupapashwe ngoLwesithathu kwi-ACS Publications, luchaza indlela enobucayi, enye inyathelo lokuhlalutya ngokukhawuleza i-viral nucleic acids kunye nokubonakala kwabo usebenzisa iiprotheyini ezikhanyayo eziluhlaza okwesibhakabhaka okanye eziluhlaza.
Ukuchongwa kwe-pathogens ngokufumanisa iminwe yabo ye-nucleic acid sisicwangciso esibalulekileyo ekuxilongweni kweklinikhi, uphando lwe-biomedical, kunye nokutya kunye nokujonga ukhuseleko lokusingqongileyo.Uvavanyo olusetyenziswa ngokubanzi lwepolymerase chain reaction (PCR) lunovakalelo oluphezulu, kodwa lufuna ukulungiswa kwesampulu ephucukileyo okanye ukutolikwa kweziphumo, oko kuzenza zingenzeki kwezinye iindawo zokhathalelo lwempilo okanye useto olunyiniweyo lwezibonelelo.
Eli qela elivela eNetherlands liyisiphumo sentsebenziswano phakathi kweenzululwazi ezivela kwiiyunivesithi kunye nezibhedlele ukuphuhlisa indlela yokuxilonga ngokukhawuleza, ephathekayo kunye nelula ukuyisebenzisa i-nucleic acid enokuthi isetyenziswe kwiindawo ezahlukeneyo.
Baphenjelelwa kukukhanya kwekhanyikhanyi, ukukhanya kwenkanyezi, kunye neenkwenkwezi ezincinci ze-aquatic phytoplankton, zonke zixhaswa yinto ebizwa ngokuba yi-bioluminescence.Oku kukhanya-ebumnyameni kubangelwa ukusabela kweekhemikhali ezibandakanya iprotheni ye-luciferase.Izazinzulu zifake iiprotheyini ze-luciferase kwi-sensors ezikhupha ukukhanya ukuququzelela ukujonga xa zifumana itekeni.Ngelixa oku kusenza ukuba ezi zivamvo zilungele ukuchongwa kwindawo yokhathalelo, okwangoku ziswele ubuntununtunu obuphezulu obufunekayo kuvavanyo loxilongo lweklinikhi.Ngelixa indlela yokuhlela i-gene ye-CRISPR inokubonelela ngobu buchule, ifuna amanyathelo amaninzi kunye nezixhobo ezongezelelweyo ezikhethekileyo zokubona umqondiso obuthathaka onokuthi ube khona kwiisampuli ezinzima, ezinomsindo.
Abaphandi baye bafumanisa indlela yokudibanisa iprotheni ehambelana ne-CRISPR kunye nesignali ye-bioluminescent enokuthi ibonwe ngekhamera yedijithali elula.Ukuqinisekisa ukuba kukho i-RNA eyaneleyo okanye isampuli ye-DNA yohlalutyo, abaphandi benza i-recombinase polymerase amplification (RPA), ubuchule obulula obusebenza kwiqondo lokushisa elingaguqukiyo malunga ne-100 ° F.Baye baphuhlisa iqonga elitsha elibizwa ngokuba yiLuminescent Nucleic Acid Sensor (LUNAS), apho iiprotheni ezimbini ze-CRISPR/Cas9 zithe ngqo kwiinxalenye ezidityanisiweyo zentsholongwane ye-genome, nganye ineqhekeza elikhethekileyo le-luciferase elidityaniswe ngasentla.
Xa i-genome ethile yentsholongwane abaphandi bahlola ngayo, iiprotheni ezimbini ze-CRISPR / Cas9 zibophelela kulandelelwano lwe-nucleic acid ekujoliswe kuyo;ziba kufutshane, zivumela iprotheyini ye-luciferase engaguqukiyo ukuba yenze kwaye ikhuphe ukukhanya okuluhlaza okwesibhakabhaka phambi kwe-substrate yekhemikhali..Ukuphendula nge-substrate esetyenzisiweyo kule nkqubo, abaphandi basebenzisa impendulo yokulawula ekhupha ukukhanya okuluhlaza.Ityhubhu etshintsha umbala ukusuka eluhlaza ukuya eluhlaza okwesibhakabhaka ibonisa umphumo omuhle.
Abaphandi bavavanya iqonga labo ngokuphuhlisa uvavanyo lwe-RPA-LUNAS, olubonayoI-SARS-CoV-2 RNAngaphandle kokwahlukaniswa kwe-RNA edinayo, kwaye yabonisa ukusebenza kwayo kokuxilonga kwiisampulu ze-nasopharyngeal swab ezivelaI-COVID 19izigulane.I-RPA-LUNAS ichonge ngempumelelo i-SARS-CoV-2 kwisithuba semizuzu engama-20 kwiisampulu ezinomthamo wentsholongwane egazini we-RNA ophantsi njengeekopi ezingama-200/μL.
Abaphandi bakholelwa ukuba uvavanyo lwabo lunokufumanisa ngokulula nangempumelelo ezinye iintsholongwane ezininzi."I-RPA-LUNAS inomtsalane kuvavanyo lwesifo esosulelayo," babhala.