Abacwaningi baseDashi bahlanganisa i-CRISPR kanye ne-bioluminescence ekuhlolweni kokuhlolaizifo ezithathelwanayo

Iphrotheni esanda kwakhiwa yasebusuku ingasheshisa futhi yenze kube lula ukutholakala kwezifo ezibangelwa amagciwane, ngokusho kwabacwaningi baseNetherlands.
Ucwaningo lwabo, olushicilelwe ngoLwesithathu ku-ACS Publications, luchaza indlela ebucayi, enesinyathelo esisodwa sokuhlaziya ngokushesha i-viral nucleic acid kanye nokubukeka kwayo kusetshenziswa amaprotheni akhanyayo aluhlaza okwesibhakabhaka noma aluhlaza.
Ukuhlonzwa kwamagciwane ngokuthola izigxivizo zeminwe ze-nucleic acid kuyisu elibalulekile ekuxilongweni komtholampilo, ucwaningo lwe-biomedical, kanye nokuqapha kokudla kanye nokuphepha kwemvelo.Ukuhlola okusetshenziswa kakhulu kwe-quantitative polymerase chain reaction (PCR) kuzwela kakhulu, kodwa kudinga ukulungiswa kwesampula okuyinkimbinkimbi noma ukuchazwa kwemiphumela, okuyenza ingasebenzi kwezinye izilungiselelo zokunakekelwa kwezempilo noma izilungiselelo ezikhawulelwe yinsiza.
Leli qembu elivela eNetherlands liwumphumela wokubambisana phakathi kososayensi abavela emanyuvesi nasezibhedlela ukuthuthukisa indlela yokuxilonga esheshayo, ephathekayo nesebenziseka kalula ye-nucleic acid engasetshenziswa ezindaweni ezihlukahlukene.
Zagqugquzelwa ukukhanya kokhanyikhanyi, ukukhanya kokhanyikhanyi, nezinkanyezi ezincane ze-aquatic phytoplankton, zonke zisekelwa into ebizwa ngokuthi i-bioluminescence.Lo mphumela wokukhanya-ebumnyameni ubangelwa ukusabela kwamakhemikhali okubandakanya iphrotheni ye-luciferase.Ososayensi bahlanganisa amaprotheni e-luciferase kuzinzwa ezikhipha ukukhanya ukuze kube lula ukubhekwa lapho zithola okuhlosiwe.Nakuba lokhu kwenza lezi zinzwa zilungele ukutholwa kwendawo yokunakekelwa, okwamanje azinakho ukuzwela okuphezulu okudingekayo ekuhlolweni kokuxilonga komtholampilo.Nakuba indlela yokuhlela yofuzo ye-CRISPR inganikeza leli khono, idinga izinyathelo eziningi kanye nezinto zokusebenza ezikhethekile ezengeziwe ukuze kutholwe isignali ebuthakathaka engaba khona kumasampuli ayinkimbinkimbi, anomsindo.
Abacwaningi bathole indlela yokuhlanganisa iphrotheni ehlobene ne-CRISPR nesignali ye-bioluminescent engatholwa ngekhamera yedijithali elula.Ukuqinisekisa ukuthi kunesampula eyanele ye-RNA noma ye-DNA ukuze ihlaziywe, abacwaningi benze i-recombinase polymerase amplification (RPA), indlela elula esebenza ezingeni lokushisa elingaguquki elingu-100°F.Bakha inkundla entsha ebizwa ngokuthi i-Luminescent Nucleic Acid Sensor (LUNAS), lapho amaprotheni amabili e-CRISPR/Cas9 eqondene ngqo nezingxenye ezihlukene ze-genome yegciwane, ngayinye inesiqephu esiyingqayizivele se-luciferase esinamathiselwe kuzo ngenhla.
Uma i-genome ethile yegciwane abaphenyi abahlolayo ikhona, amaprotheni amabili e-CRISPR/Cas9 abopha ngokulandelana okuhlosiwe kwe-nucleic acid;ziba seduze, zivumela iphrotheni ye-luciferase engaguquki ukuba yakhe futhi ikhiphe ukukhanya okuluhlaza okwesibhakabhaka phambi kwe-substrate yamakhemikhali..Ukulandisa nge-substrate edliwe kule nqubo, abacwaningi basebenzisa ukusabela kokulawula okukhipha ukukhanya okuluhlaza.Ishubhu elishintsha umbala ukusuka koluhlaza kuya kokuluhlaza okwesibhakabhaka libonisa umphumela omuhle.
Abacwaningi bahlole inkundla yabo ngokwakha i-RPA-LUNAS assay, etholaI-SARS-CoV-2 RNAngaphandle kokuhlukaniswa kwe-RNA eyisicefe, futhi yabonisa ukusebenza kwayo kokuxilonga kumasampula e-nasopharyngeal swab kusukaI-COVID 19iziguli.I-RPA-LUNAS ithole ngempumelelo i-SARS-CoV-2 phakathi nemizuzu engama-20 kumasampula anenani legciwane le-RNA eliphansi njengamakhophi angu-200/μL.
Abacwaningi bakholelwa ukuthi ukuhlola kwabo kungathola kalula nangempumelelo amanye amagciwane amaningi."I-RPA-LUNAS iyathandeka ekuhlolweni kwezifo ezithathelwanayo," babhala.