Uvavanyo lwe-DNA methylation ludityaniswe nee-smartphones zokuhlolwa kwangaphambili kwe-tumor kunye nokuhlolwa kwe-leukemia ngokuchaneka kwe-90.0%!

Ukufunyaniswa kwangoko komhlaza ngokusekwe kwi-biopsy yolwelo yindlela entsha yokufunyaniswa komhlaza kunye noxilongo olucetywe liZiko loMhlaza leSizwe lase-US kwiminyaka yakutshanje, ngenjongo yokufumanisa umhlaza kwangethuba okanye izilonda ezingaphambi komhlaza. Isetyenziswe ngokubanzi njengenoveli biomarker yokuxilongwa kwangoko kwezifo ezahlukeneyo, kubandakanya umhlaza wemiphunga, amathumba emathunjini, igliomas kunye namathumba e-gynecological.

Ukuvela kwamaqonga okuchonga i-methylation landscape (Methylscape) biomarkers inamandla okuphucula kakhulu uhlolo olukhoyo lwakwangoko lomhlaza, ukubeka izigulane kwinqanaba elinyangekayo.

Kutshanje, abaphandi baye baphuhlisa iqonga lokuva elilula nelithe ngqo lokufunyanwa komhlaba we-methylation esekwe kwi-cysteamine ehonjisiweyo ye-nanoparticles yegolide (Cyst/AuNPs) edityaniswe ne-biosensor esekwe kwi-smartphone eyenza ukuba kuhlolwe ngokukhawuleza kwangethuba uluhlu olubanzi lwamathumba. Ukuhlolwa kwangaphambili kwe-leukemia kunokwenziwa ngaphakathi kwemizuzu eyi-15 emva kokutsalwa kwe-DNA kwisampulu yegazi, ngokuchaneka kwe-90.0%. Isihloko senqaku kukufunyaniswa ngokukhawuleza komhlaza we-DNA egazini lomntu usebenzisa i-cysteamine-capped AuNPs kunye ne-smartphone yokufunda ngomatshini.

Umzobo 1. Iqonga elilula nelikhawulezayo lokubona umhlaza ngokusetyenziswa kweCyst/AuNPs components inokufezekiswa ngamanyathelo amabini alula.

Oku kuboniswe kuMzobo 1. Okokuqala, isisombululo esinamanzi sasisetyenziselwa ukutshabalalisa iziqwenga zeDNA. I-Cyst / AuNPs zongezwa kwisisombululo esixubileyo. I-DNA eqhelekileyo kunye ne-malignant ineempawu ezahlukeneyo ze-methylation, ezikhokelela kwiinqununu ze-DNA ezineendlela ezahlukeneyo zokuzihlanganisa. I-DNA eqhelekileyo idibanisa ngokukhululekileyo kwaye ekugqibeleni idibanisa i-Cyst / AuNPs, ebangela ukuba i-red-shifted nature of Cyst / AuNPs, ukwenzela ukuba utshintsho kumbala obomvu ukuya kumfusa lunokubonwa ngeso lenyama. Ngokwahlukileyo, iprofayili ekhethekileyo ye-methylation yomhlaza weDNA ikhokelela ekuvelisweni kwamaqela amakhulu amaqhekeza eDNA.

Imifanekiso yeepleyiti ezinamaqula angama-96 zithathwe kusetyenziswa ikhamera ye-smartphone. ICancer DNA yalinganiswa nge-smartphone exhotyiswe ngoomatshini bokufunda xa kuthelekiswa neendlela ezisekwe kwi-spectroscopy.

Ukuhlolwa komhlaza kwiisampuli zegazi lokwenyani

Ukwandisa ukusetyenziswa kweqonga lokuva, abaphandi basebenzise inzwa eyahlula ngempumelelo phakathi kweDNA eqhelekileyo kunye nomhlaza kwiisampuli zegazi zangempela. iipateni ze-methylation kwiindawo ze-CpG zilawula i-gene expression. Phantse kuzo zonke iintlobo zomhlaza, utshintsho kwi-DNA methylation kwaye ngaloo ndlela ekubonakalisweni kwezakhi zofuzo ezikhuthaza i-tumourigenesis ziye zabonwa ukuba zitshintshe.

Njengomzekelo wezinye ii-cancer ezinxulumene ne-DNA methylation, abaphandi basebenzise iisampulu zegazi kwizigulane ze-leukemia kunye nolawulo olunempilo ukuphanda ukusebenza kwe-methylation landscape ekwahluleni i-leukemic cancers. Le biomarker ye-methylation landscape ayigqithisi kuphela iindlela ezikhoyo zokuhlola i-leukemia ekhawulezayo, kodwa ikwabonisa ukuba nokwenzeka kokwandisa ekubhaqweni kwangaphambili kweentlobo ezininzi zomhlaza kusetyenziswa olu vavanyo lulula noluthe ngqo.

I-DNA evela kwiisampulu zegazi kwizigulane ze-leukemia ezingama-31 kunye nabantu abasempilweni abali-12 bahlalutywa. njengoko kuboniswe kwibhokisi yebhokisi kwi-Figure 2a, i-absorbection ehambelanayo yeesampuli zomhlaza (ΔA650 / 525) yayingaphantsi kune-DNA evela kwiisampuli eziqhelekileyo. oku bekungenxa yophuculo lwe-hydrophobicity ekhokelela ekudibaneni komhlaza weDNA, ethintele ukudityaniswa kweCyst/AuNPs. Ngenxa yoko, ezi nanoparticles zahlakazwa ngokupheleleyo kwiingqimba zangaphandle ze-aggregates zomhlaza, ezibangele ukusabalalisa okwahlukileyo kwe-Cyst / AuNPs adsorbed kwizinto eziqhelekileyo kunye nomhlaza we-DNA aggregates. Iijika ze-ROC zaye zenziwa ngokutshintsha umda ukusuka kwixabiso elincinci le-ΔA650 / 525 ukuya kwixabiso eliphezulu.

Umzobo 2.(a) Amaxabiso anxulumene ne-cyst / AuNPs abonisa ubukho besiqhelo (blue) kunye nomhlaza (obomvu) we-dna phantsi kweemeko eziphuculweyo.

(DA650/525) yeebhokisi zebhokisi; (b) Uhlalutyo lwe-ROC kunye novavanyo lweemvavanyo zokuxilonga. (c) I-matrix yokudideka yokuxilongwa kwezigulane eziqhelekileyo kunye nomhlaza. (d) Uvakalelo, ukuchaneka, ixabiso elihle lokuxela kwangaphambili (PPV), ixabiso elibi lokuxela kwangaphambili (NPV) kunye nokuchaneka kwendlela yokuphuhliswa.

Njengoko kuboniswe kwi-Figure 2b, indawo ephantsi kwe-ROC curve (AUC = 0.9274) efunyenwe kwi-sensor ephuhlisiweyo ibonise uvakalelo oluphezulu kunye neenkcukacha. Njengoko kunokubonwa kwibhokisi yebhokisi, indawo ephantsi emele iqela le-DNA eqhelekileyo ayihlulwe kakuhle kwindawo ephakamileyo emele iqela le-DNA yomhlaza; ngoko ke, ukuguqulwa kwezinto kusetyenziswe ukwahlula phakathi kwamaqela aqhelekileyo kunye nomhlaza. Ngokunikezelwa kweenguqu ezizimeleyo, iqikelela ukuba kunokwenzeka ukuba kwenzeke isiganeko, esifana nomhlaza okanye iqela eliqhelekileyo. Uluhlu oluxhomekeke kuluhlu phakathi kwe-0 kunye ne-1. Isiphumo ke ngoko yinto enokwenzeka. Sinqume ukuba nokwenzeka kokuchongwa komhlaza (P) ngokusekelwe kwi-ΔA650/525 ngolu hlobo lulandelayo.

apho b=5.3533,w1=-6.965. Ukwahlula isampuli, ithuba elingaphantsi kwe-0.5 libonisa isampuli eqhelekileyo, ngelixa i-0.5 okanye ngaphezulu ibonisa isampuli yomhlaza. Umzobo we-2c ubonisa i-matrix yokudideka eyenziwe kwi-level-yodwa-cross-validation, eyayisetyenziselwa ukuqinisekisa ukuzinza kwendlela yokwahlula. Umzobo we-2d ushwankathela uvavanyo lokuxilonga uvavanyo lwendlela, kubandakanywa uvakalelo, ukuchaneka, ixabiso eliqikelelweyo eliqinisekileyo (PPV) kunye nexabiso elibi lokuxela kwangaphambili (NPV).

I-biosensors esekwe kwi-smartphone

Ukwenza lula ngakumbi uvavanyo lwesampulu ngaphandle kokusebenzisa i-spectrophotometers, abaphandi basebenzisa ubukrelekrele bokwenziwa (AI) ukutolika umbala wesisombululo kunye nokwahlula phakathi kwabantu abaqhelekileyo kunye nomhlaza. Ukunikezelwa oku, umbono wekhompyutheni wawusetyenziselwa ukuguqulela umbala wesisombululo seCyst / AuNPs kwi-DNA eqhelekileyo (obomvu) okanye i-DNA enomhlaza (obomvu) usebenzisa imifanekiso ye-96-well plates ethathwe ngekhamera yeselula. Ubukrelekrele bokwenziwa bunokunciphisa iindleko kunye nokuphucula ukufikeleleka ekutolikeni umbala wezisombululo ze-nanoparticle, kwaye ngaphandle kokusetyenziswa kwezixhobo ze-smartphone ze-optical hardware. Ekugqibeleni, iimodeli ezimbini zokufunda koomatshini, kubandakanywa iHlathi eliHlangeneyo (RF) kunye ne-Support Vector Machine (SVM) baqeqeshelwa ukwakha iimodeli. zombini iimodeli ze-RF kunye ne-SVM zihlele ngokufanelekileyo iisampuli njenge-positive kunye ne-negative ngokuchaneka kwe-90.0%. Oku kuphakamisa ukuba ukusetyenziswa kobukrelekrele bokwenziwa kwifowuni esekwe kwi-biosensing kunokwenzeka.

Umzobo 3. (a) Iklasi ekujoliswe kuyo yesisombululo esirekhodiweyo ngexesha lokulungiswa kwesampuli yesinyathelo sokufumana umfanekiso. (b) Umzekelo womfanekiso othathwe ngexesha lenyathelo lokufumana umfanekiso. (c) Ukuqina kombala we-cyst / AuNPs isisombululo kwiqula ngalinye le-96-well plate ekhutshwe kumfanekiso (b).

Ukusebenzisa i-Cyst / AuNPs, abaphandi baye baphuhlisa ngempumelelo i-platform ye-sensing elula yokubona indawo ye-methylation kunye ne-sensor ekwazi ukwahlula i-DNA eqhelekileyo kwi-DNA yomhlaza xa usebenzisa iisampuli zegazi zangempela zokuhlola i-leukemia. I-sensor ephuhlisiwe ibonise ukuba i-DNA ekhutshwe kwiisampuli zegazi zangempela yakwazi ukufumanisa ngokukhawuleza kunye neendleko-ngexabiso ixabiso elincinci lomhlaza we-DNA (3nM) kwizigulane ze-leukemia kwimizuzu ye-15, kwaye ibonise ukuchaneka kwe-95.3%. Ukwenza lula ngakumbi uvavanyo lwesampulu ngokuphelisa imfuno ye-spectrophotometer, ukufundwa komatshini kwasetyenziselwa ukutolika umbala wesisombululo kunye nokwahlula phakathi kwabantu abaqhelekileyo kunye nomhlaza usebenzisa ifoto yefowuni ephathwayo, kwaye ukuchaneka kuye kwafikelelwa kwi-90.0%.

Isalathiso: DOI: 10.1039/d2ra05725e