Ukuhlolwa kwe-DNA methylation kuhlanganiswe nama-smartphones ukuze kuhlolwe amathumba kusenesikhathi kanye nokuhlolwa kwe-leukemia ngokunemba okungu-90.0%!

Ukutholwa komdlavuza kusenesikhathi okususelwa ku-liquid biopsy kuyindlela entsha yokutholwa nokuxilongwa komdlavuza ehlongozwa yi-US National Cancer Institute eminyakeni yamuva nje, ngenhloso yokuthola umdlavuza ngaphambi kwesikhathi noma izilonda eziyingozi. Isetshenziswe kabanzi njenge-biomarker yenoveli yokuxilongwa kusenesikhathi kwezifo ezihlukahlukene ezibulalayo, okuhlanganisa umdlavuza wamaphaphu, izimila zesisu, ama-gliomas kanye nezimila ze-gynecological.

Ukuvela kwamapulatifomu okuhlonza ama-biomarker e-methylation landscape (Methylscape) anamandla okuthuthukisa kakhulu ukuhlolelwa umdlavuza okukhona kakade, okubeka iziguli esigabeni esingelaphekayo sokuqala.

Muva nje, abacwaningi bathuthukise inkundla yokuzwa elula neqondile yokutholwa kwe-methylation landscape esekelwe ku-cysteamine ehlotshiswe ngama-nanoparticles egolide (Cyst/AuNPs) ahlanganiswe ne-biosensor esekelwe ku-smartphone eyenza ukuhlolwa kwangaphambi kwesikhathi kwezinhlobo eziningi zezimila. Ukuhlolwa kusenesikhathi kwe-leukemia kungenziwa phakathi nemizuzu engu-15 ngemva kokukhishwa kwe-DNA esithathweni segazi, ngokunemba okungama-90.0%. Isihloko se-athikili sithi Ukutholwa Ngokushesha kwe-DNA yomdlavuza egazini lomuntu kusetshenziswa ama-AuNP ane-cysteamine-capped kanye ne-smartphone enikwe amandla ukufunda ngomshini.

Umfanekiso 1. Inkundla yokuzwa elula nesheshayo yokuhlolwa komdlavuza kusetshenziswa izingxenye ze-Cyst/AuNPs ingafezwa ngezinyathelo ezimbili ezilula.

Lokhu kuboniswa kuMfanekiso 1. Okokuqala, isixazululo esinamanzi sasetshenziselwa ukuhlakaza izingcezu ze-DNA. I-Cyst/AuNPs yabe yengezwa kusixazululo esixubile. I-DNA evamile neyingozi inezinto ezihlukahlukene ze-methylation, okuholela ezingcezu ze-DNA ezinamaphethini ahlukene okuzihlanganisa. I-DNA evamile ihlanganisa ngokukhululekile futhi ekugcineni ihlanganise i-Cyst/AuNPs, okuholela ekushintsheni okubomvu kwemvelo ye-Cyst/AuNPs, ukuze ukuguqulwa kombala osuka kobomvu ukuya kokunsomi kubonwe ngeso lenyama. Ngokuphambene, iphrofayili ye-methylation eyingqayizivele yomdlavuza we-DNA iholela ekukhiqizweni kwamaqoqo amakhulu wezingcezu ze-DNA.

Izithombe zamapuleti anemithombo engu-96 zithathwe kusetshenziswa ikhamera ye-smartphone. ICancer DNA ikalwa nge-smartphone efakwe ukufunda ngomshini uma iqhathaniswa nezindlela ezisekelwe ku-spectroscopy.

Ukuhlolwa komdlavuza kumasampula egazi langempela

Ukuze banwebe ukusetshenziswa kwenkundla yezinzwa, abaphenyi basebenzise inzwa ehlukanise ngempumelelo phakathi kwe-DNA evamile nenomdlavuza kumasampula egazi langempela. amaphethini e-methylation kumasayithi e-CpG alawula nge-epigenetically ukubonakaliswa kofuzo. Cishe kuzo zonke izinhlobo zomdlavuza, izinguquko ku-DNA methylation futhi ngaleyo ndlela ekukhulumeni kwezakhi zofuzo ezikhuthaza i-tumourigenesis kuye kwabonwa ukushintshana.

Njengemodeli yezinye izinhlobo zomdlavuza ezihlotshaniswa ne-DNA methylation, abacwaningi basebenzisa amasampula egazi ezigulini ze-leukemia kanye nezilawuli ezinempilo ukuze baphenye ukusebenza kahle kwe-methylation landscape ekuhlukaniseni umdlavuza we-leukemic. Le biomarker ye-methylation landscape ayiphumeleli nje kuphela izindlela ezikhona zokuhlola i-leukemia esheshayo, kodwa futhi ikhombisa ukuba nokwenzeka kokunwetshwa kokutholwa kusenesikhathi kwezinhlobo eziningi zomdlavuza kusetshenziswa lolu cwaningo olulula noluqondile.

Kwahlaziywa i-DNA evela kumasampula egazi ezigulini ezingama-31 ze-leukemia kanye nabantu abangu-12 abanempilo. njengoba kuboniswe esakhiweni sebhokisi ku-Figure 2a, ukumuncwa okuhlobene kwamasampula omdlavuza (ΔA650/525) kwakungaphansi kwalokho kwe-DNA evela kumasampuli avamile. lokhu kwakungenxa yokuthuthukiswa kwe-hydrophobicity okuholela ekuhlanganisweni okuminyene kwe-DNA yomdlavuza, okuvimbele ukuhlangana kwe-Cyst/AuNPs. Ngenxa yalokho, lawa ma-nanoparticles ahlakazeke ngokuphelele ezingqimbeni zangaphandle ze-aggregate yomdlavuza, okuholele ekuhlakazweni okuhlukile kwe-Cyst/AuNPs ekhangiswe kuma-aggregate we-DNA omdlavuza. Amajika e-ROC abe esekhiqizwa ngokushintsha i-threshold ukusuka kunani elincane le-ΔA650/525 ukuya kunani eliphezulu.

Umfanekiso 2.(a) Amanani okumunca ahlobene ezixazululo ze-cyst/AuNPs ezibonisa ukuba khona kwe-dna evamile (eluhlaza okwesibhakabhaka) kanye nomdlavuza (obomvu) ngaphansi kwezimo ezithuthukisiwe.

(DA650/525) yeziza zamabhokisi; (b) Ukuhlaziywa kwe-ROC nokuhlolwa kokuhlolwa kokuxilonga. (c) I-matrix yokudideka yokuxilonga iziguli ezivamile nezinomdlavuza. (d) Ukuzwela, ukucacisa, inani elihle lokubikezela (PPV), inani elibi lokubikezela (NPV) kanye nokunemba kwendlela ethuthukisiwe.

Njengoba kuboniswe ku-Figure 2b, indawo engaphansi kwejika le-ROC (AUC = 0.9274) etholwe inzwa ethuthukisiwe ibonise ukuzwela okuphezulu nokucacisa. Njengoba kungabonakala kusakhiwo sebhokisi, iphuzu eliphansi kunazo zonke elimelela iqembu le-DNA elivamile alihlukanisiwe kahle nephuzu eliphakeme elimelela iqembu le-DNA yomdlavuza; ngakho-ke, ukuhlehliswa kwezinto ezibonakalayo kwasetshenziselwa ukuhlukanisa phakathi kwamaqembu avamile kanye nomdlavuza. Ngokunikezwa kwesethi yokuhlukahluka okuzimele, ilinganisela amathuba okuba kwenzeke umcimbi, njengomdlavuza noma iqembu elivamile. Ububanzi obuncikile buphakathi kuka-0 no-1. Ngakho-ke umphumela ungamathuba. Sinqume amathuba okuhlonza umdlavuza (P) ngokusekelwe ku-ΔA650/525 kanje.

lapho b=5.3533,w1=-6.965. Ngokuhlukaniswa kwesampula, amathuba angaphansi kuka-0.5 abonisa isampula evamile, kuyilapho amathuba angu-0.5 noma ngaphezulu abonisa isampula yomdlavuza. Umfanekiso 2c ubonisa i-matrix yokudideka ekhiqizwe kusukela ekuqinisekiseni ukushiya yodwa, okusetshenziswe ukuze kuqinisekiswe ukuzinza kwendlela yokuhlukanisa. Umfanekiso we-2d ufingqa ukuhlolwa kokuhlolwa kokuxilonga kwendlela, okuhlanganisa ukuzwela, ukucaciswa, inani elihle lokubikezela (PPV) kanye nevelu yokubikezela engalungile (NPV).

Ama-biosensors asuselwa ku-smartphone

Ukuze kwenziwe lula ukuhlolwa kwesampula ngaphandle kokusebenzisa ama-spectrophotometers, abacwaningi basebenzisa ubuhlakani bokwenziwa (AI) ukuze bahumushe umbala wesixazululo futhi bahlukanise phakathi kwabantu abavamile nabanomdlavuza. Njengoba kunikezwe lokhu, umbono wekhompyutha wasetshenziselwa ukuhumusha umbala wesixazululo se-Cyst/AuNPs ube yi-DNA evamile (onsomi) noma i-DNA enomdlavuza (obomvu) kusetshenziswa izithombe zamapuleti angama-96 athathwe ngekhamera yeselula. Ubuhlakani bokwenziwa bunganciphisa izindleko futhi buthuthukise ukufinyeleleka ekuchazeni umbala wezixazululo ze-nanoparticle, futhi ngaphandle kokusebenzisa noma yiziphi izesekeli ze-smartphone yehadiwe ye-optical. Ekugcineni, amamodeli wokufunda womshini amabili, okuhlanganisa i-Random Forest (RF) kanye ne-Support Vector Machine (SVM) aqeqeshelwa ukwakha amamodeli. womabili amamodeli e-RF kanye ne-SVM ahlukanise ngokufanelekile amasampuli njengalihle nelibi ngokunemba okungu-90.0%. Lokhu kuphakamisa ukuthi ukusetshenziswa kobuhlakani bokwenziwa ekutholeni i-biosensing esekelwe kumakhalekhukhwini kungenzeka.

Umfanekiso 3.(a) Isigaba esiqondisiwe sesixazululo esirekhodwe ngesikhathi sokulungiswa kwesampula yesinyathelo sokutholwa kwesithombe. (b) Isibonelo sesithombe esithathwe phakathi nesinyathelo sokutholwa kwesithombe. (c) Ukuqina kombala kwesixazululo se-cyst/AuNPs emthonjeni ngamunye wepuleti lemithombo engu-96 elikhishwe esithombeni (b).

Besebenzisa i-Cyst/AuNPs, abacwaningi baye bakha ngempumelelo inkundla yokuzwa elula yokutholwa kwe-methylation landscape kanye nenzwa ekwazi ukuhlukanisa i-DNA evamile ku-DNA yomdlavuza lapho kusetshenziswa amasampula egazi langempela ukuze kuhlolwe i-leukemia. Inzwa ethuthukisiwe ibonise ukuthi i-DNA ekhishwe kumasampula egazi langempela ikwazile ukusheshe ibone amanani amancane e-DNA yomdlavuza (3nM) ezigulini ze-leukemia emizuzwini engu-15, futhi yabonisa ukunemba okungu-95.3%. Ukuze kwenziwe lula ukuhlolwa kwesampula ngokuqeda isidingo se-spectrophotometer, ukufundwa komshini kwasetshenziswa ukuze kuhunyushwe umbala wesixazululo futhi kuhlukanise phakathi kwabantu abavamile nabanomdlavuza kusetshenziswa isithombe sikamakhalekhukhwini, futhi ukunemba nakho kwatholakala ku-90.0%.

Inkomba: DOI: 10.1039/d2ra05725e