Amakuru

Ingamba gakondo zo kwisuzumisha mugutahura indwara zandura zisaba gukoresha ibikoresho byintebe bidakwiriye kwipimisha ingingo (POCT).Microfluidics ivuka ni tekinoroji ya miniaturike, yikora, kandi ihuriweho hamwe nuburyo bushobora gukoreshwa muburyo bwa gakondo bwo kwihuta, bidahenze, byukuri kwisuzumisha kurubuga.Uburyo bwo gusuzuma indwara ya molekulari bukoreshwa cyane mubikoresho bya microfluidic nkuburyo bwiza cyane bwo kumenya indwara.Iri suzuma ryerekana muri make iterambere ryagezweho muri microfluidic ishingiye kuri molekuline yo gusuzuma indwara zandura haba mu myigire no mu nganda.Ubwa mbere, turasobanura uburyo busanzwe bwo gutunganya acide nucleic, harimo kwigana icyitegererezo, kwongera imbaraga, no gusoma ibimenyetso.Ibiranga, ibyiza nibibi byubwoko bune bwa microfluidic noneho bigereranywa.Ibikurikira, tuzaganira ku ikoreshwa rya sisitemu yo kubara kwinshi kwa acide nucleic.Byombi bya kera na vuba byubucuruzi microfluidic ishingiye kubikoresho byo gusuzuma bya molekile byapimwe muri make nkibimenyetso byerekana uko isoko rihagaze.Hanyuma, turasaba icyerekezo kizaza cyo gusuzuma microfluidic yo gusuzuma indwara zanduza.
Indwara zandura ziterwa na virusi, harimo bagiteri, virusi, na parasite, zikwirakwizwa ku isi.Bitandukanye n’izindi ndwara, virusi zandura vuba kandi zikwirakwira hagati y’abantu n’inyamaswa zakira binyuze mu gukingira, itangazamakuru ry’ikirere n’amazi [1].Kwirinda indwara zandura ni ngombwa nkigipimo cyubuzima rusange.Ingamba eshatu zingenzi zo kurwanya indwara zanduza: (1) kugenzura inkomoko yanduye;(2) guhagarika inzira yo kohereza;(3) kurinda abaturage byoroshye.Mu ngamba zingenzi, kugenzura inkomoko yanduye bifatwa nkingamba zingenzi bitewe nuburyo bworoshye nigiciro gito.Gusuzuma byihuse, kwigunga, no kuvura abantu banduye ni ingenzi, bisaba ingamba zihuse zo kwisuzumisha [2].Kugeza ubu isuzuma ry'indwara zandura ubusanzwe rihuza ibizamini byo kwa muganga bishingiye ku bimenyetso n'ibimenyetso ndetse n'ubushakashatsi bwa laboratoire nk'umuco w'akagari ndetse no gusuzuma indwara ya molekile, bisaba abakozi bahuguwe, uburyo bukoreshwa cyane n'umurimo, n'ibikoresho bihenze byo gupima [3, 4].Kwirinda icyorezo cyandura bisaba kwisuzumisha byihuse, bidahenze, kandi neza, cyane cyane mu turere dufite amikoro make aho indwara zandura zikunze kugaragara kandi zikomeye [5], ndetse no kuvurwa mu butayu cyangwa ku rugamba, aho ibintu byihutirwa bitateganijwe..ubuvuzi bufite aho bugarukira [6].Ni muri urwo rwego, microfluidics ni ikoranabuhanga rihuza tekinoroji ya mikorobe ya elegitoroniki, nanotehnologiya, cyangwa ibikoresho bya siyansi yo gukoresha neza amazi [7,8,9,10], bitanga uburyo bushya bwo kumenya ingingo-yitaweho (POCT).) imiti yandura hanze yibitaro na laboratoire.Ugereranije no kwisuzumisha bisanzwe bitwara igihe, tekinoroji ya microfluidic itanga icyitegererezo hamwe no kuzigama amafaranga yo gusuzuma molekile mugihe cyo kwandura indwara.Ikwirakwizwa ry’indwara ya coronavirus ku isi hose (COVID-19) riterwa na syndrome ikabije y’ubuhumekero coronavirus 2 (SARS-CoV-2), bityo akamaro ka microfluidics mu gukumira no kurwanya icyorezo ku gihe cyongeye gushimangirwa [11, 12 , 13].Bitandukanye no kwisuzumisha gakondo, microfluidic POCT ikoresha ibikoresho bito byimukanwa kuva kubisesengura intebe kugeza ku bice bito byo kwipimisha kugirango bipimishe hafi y'icyitegererezo [14].Ibi bizamini biranga imyiteguro yoroshye cyangwa ntanicyitegererezo, kwihutisha ibimenyetso byihuse, hamwe no gusoma ibimenyetso byoroshye bivamo igihe gito nibisubizo nyabyo muminota mike.Kuboneka no gutanga umusaruro mwinshi wibikoresho byubuvuzi bishingiye kuri microfluidic byaguye uburyo bwabo bwo kwisuzumisha buhendutse kandi butaziguye hanze yibitaro, hafi yumurwayi, ndetse no murugo.
Mu ngamba zihari zo gusuzuma indwara zandura, gusuzuma molekile ni imwe mu zumva cyane [15, 16].Byongeye kandi, kwisuzumisha kuri molekuline akenshi bikoreshwa nkurwego rwa zahabu mugukomeza gutahura COVID-19, bigatuma habaho kumenya mu buryo butaziguye uturere twihariye twa virusi ya RNA cyangwa ADN mbere yuko igisubizo cy’ubudahangarwa [17, 18].Mu isubiramo ryubu, turerekana iterambere rigezweho muri microfluidics ishingiye kuri molekuline yo gusuzuma indwara zandura, duhereye ku myigire kugeza ku nganda zizaza (Ishusho 1).Tuzatangirana nintambwe eshatu zingenzi mugushakisha acide nucleic: kuri chip chip sample prereatment, amplification acide nucleic, no gusoma ibimenyetso.Hanyuma twagereranije ubwoko butandukanye bwa microfluidic platform nuburyo n'imikorere yabyo, twerekana ibiranga bidasanzwe (imbaraga nintege nke).Kumenyekanisha aside nucleic aside irasuzumwa kandi igatangwa nkurugero rwikoranabuhanga rya gatatu kugirango habe umubare wuzuye wa molekile zanduza.Mubyongeyeho, ibikoresho byinshi bisanzwe kandi bigezweho byubucuruzi POCT bizerekanwa kugirango bigaragaze uko isoko rya microfluidic POCT ihagaze kugirango isuzume molekile.Tuzaganira kandi tunasobanure icyerekezo cyacu kubisabwa ejo hazaza.
Module ya microfluidic chip yo gutahura aside nucleic irashobora kugabanywamo ibyiciro bitatu (icyitegererezo, kumenyekana, no gutangaza) ukurikije imirimo yabo [19].Muri ubu buryo, icyitegererezo cyerekana cyane cyane urugero rwa lysis no gukuramo aside nucleic.Moderi ya sensor igenzura cyane cyane guhindura no kongera ibimenyetso bya acide nucleic.Module yerekana ibimenyetso byerekana ibimenyetso byahinduwe kandi bigakorwa na module yo kumva.Dushingiye ku nzira yo kumenya acide nucleic acide kuri chip, tuzavuga muri make chip zitandukanye zishobora kumenya imikorere "yinjiza nibisohoka".
Intambwe yambere mugushakisha aside nucleic ni ugukuramo aside nucleic, ni ukuvuga gutandukanya intego ya nucleic acide nicyitegererezo cyambere.Gukuramo aside nucleique bikorwa kugirango isukure acide nucleic iyindi myanda ihumanya, itume uburinganire bwimiterere yibanze ya molekile ya acide nucleique, kandi bunoze ibisubizo.Gukuramo aside nucleique bisaba urugero rwa lysis hamwe no gufata aside nucleic, ubwiza nubushobozi bwabyo bigira ingaruka zikomeye kubushakashatsi nibisubizo byo gusuzuma.Ingaruka mbi zose mugihe cyo gukuramo zirashobora kugabanya izindi gutahura.Kurugero, uburyo bwa polymerase reaction (PCR) hamwe nuburyo bwa loop isothermal amplification (LAMP) burahagarikwa nuburyo bumwebumwe busigaye bwumusemburo nka Ethanol na isopropanol muri reagent ya acide nucleic [20].Kuvoma amazi-kuvoma no gukuramo ibyiciro bikomeye nuburyo bukunzwe cyane bwo gutandukanya acide nucleic [21], ariko, kuvoma amazi-yamazi kuri chip ni bike cyane, kubera ko reagent zikoreshwa mugukuramo amazi-amazi bitera kwangirika kwa mikorobe nyinshi. .Hano, turagaragaza microarray ishingiye kuburyo bukomeye bwo kuvoma no kugereranya ibyiza byabo nibibi.
Silicon ni substrate yibikoresho ihujwe na acide nucleic bitewe na biocompatibilité, ituze, kandi byoroshye guhinduka [22].Icy'ingenzi, iyo ihinduwe na silika cyangwa ibindi bikoresho, iyi compte yerekana ibintu kuri adsorb acide nucleic acide nabi munsi ya pH nkeya, imiterere yumunyu mwinshi mugihe ikoresheje pH nyinshi, ibisubizo byumunyu muke.Ukurikije iki kintu, birashoboka kweza aside nucleic.
Ubwoko butandukanye bwibikoresho bishingiye kuri silika byakoreshejwe mugukuramo aside nucleic muri microfluidics, nk'amasaro ya silika, ifu, filtri ya microfibre, hamwe na silika [23, 24, 25, 26].Ukurikije imiterere yibikoresho, ibikoresho bishingiye kuri silicon birashobora gukoreshwa muri microcircuits muburyo butandukanye.Kurugero, granules ya silika, ifu, hamwe na nanofilteri yubucuruzi irashobora gushyirwa gusa mu byobo cyangwa mikorobe ya microfluidic chip kandi bigafasha gukuramo aside nucleic mu byitegererezo [27, 28, 29].Ubuso bwahinduwe na silika ya silika irashobora kandi gukoreshwa mugusukura byihuse ADN ya virusi ku giciro gito.Kurugero, Wang n'abandi.[30] Muguhuza ibyiyumvo byerekana imbaraga hamwe no guhererekanya urunigi rwa viticle-medrated membrane hamwe na silika membrane yashizwemo na chitosan oligosaccharide, hashyizweho uburyo butandukanye bwo gutwara ibintu bwerekanaga neza ibice 102-108 byakoronijwe.(CFU) / ml Vibrio parahaemolyticus., kandi kuba hari virusi byagaragaye byoroshye.Powell n'abandi.[31] Microarray zishingiye kuri Silicon zaje gukoreshwa kugirango hamenyekane virusi ya hepatite C (HCV), virusi ya immunodeficiency ya muntu (VIH), virusi ya Zika, na papillomavirus yumuntu no gukwirakwizwa mu buryo bwikora, aho hakozwe mikorobe 1,3 μl itotezwa kugira ngo ifate virusi ya RNA.hanyuma ukore muburyo bwo kwagura.Usibye ubwo buryo, microcolumns ya silika yahinduwe hejuru nayo igira uruhare runini mugukuramo aside nucleic, kuko geometrie nibintu byahinduye ibikoresho byongera cyane umusaruro.Chen n'abandi.32Iki gikoresho cya microfluidic gihuza umurongo wa 0.25 cm2 micropillars kuri substrate ya silicon kugirango ugere ku buryo bunoze bwo kuvoma binyuze mu buso bunini kugera ku gipimo cy’imibare.Ibyiza byiki gishushanyo nuko microfluidic igikoresho gishobora kugera kuri 95% yo gukuramo aside nucleic.Izi ngamba zishingiye kuri silicon zerekana agaciro ko gutandukanya acide nucleic byihuse ku giciro gito.Hamwe na microfluidic chip, ingamba zo gukuramo silikoni ntizishobora gusa kongera imikorere yo kumenya aside nucleique gusa, ahubwo inorohereza miniaturizasi no guhuza ibikoresho byisesengura [20].
Uburyo bwo gutandukanya magnetiki bukoresha ibice bya magnetique kugirango bitandukanya acide nucleic imbere yumurima wa magneti wo hanze.Ibice bya magnetiki bikoreshwa cyane birimo Fe3O4 cyangwa γ-Fe2O3 ibice bya magnetiki bisize silika, amino na carboxyl [33,34,35,36].Itandukanyirizo ryibice bya magnetiki ugereranije na silicon ishingiye kuri SPE nuburyo bworoshye bwo gukoresha no kugenzura hamwe na magneti yo hanze.
Ukoresheje imikoranire ya electrostatike hagati ya acide nucleic na silika, mugihe cyumunyu mwinshi hamwe na pH nkeya, acide nucleic irashirwa hejuru yubutaka bwa magnetiki yubatswe na silika, mugihe mubihe byumunyu muke na pH nyinshi, molekile zirashobora gukaraba na none..Amasaro ya magnetiki yubatswe na silika atuma bishoboka gukuramo ADN mubipimo binini (400 μL) ukoresheje moteri igenzurwa na rukuruzi.Nkimyiyerekano, Rodriguez-Mateos n'abandi.[38] yakoresheje magneti ahinduka kugirango agenzure ihererekanyabubasha ryamasaro mubyumba bitandukanye.Ukurikije ibice bya magnetiki bisizwe na silika, kopi 470 / mL ya SARS-CoV-2 genomic RNA irashobora gukurwa muburugero rwamazi yanduye kugirango LAMP ihindurwe (RT-LAMP) kandi igisubizo gishobora gusomwa mugihe cyisaha 1.ijisho ryambaye ubusa (Ishusho 2a).
Ibikoresho bishingiye kubikoresho bya magnetiki kandi byoroshye.Igishushanyo mbonera cya IFAST RT-LAMP microfluidic igikoresho cya SARS-CoV-2 RNA gutahura (yakuwe kuri [38]).b Igikoresho gito cya Centrifugal ya dSPE ya buccal swab nucleic aside (yakuwe muri [39]).c Yubatswe-yifashisha icyitegererezo cyibanze ukoresheje ikarita ya FTA® (yakuwe kuri [50]).d Guhuza impapuro 5 zungurura zahinduwe hamwe na chitosan (byahinduwe kuva [51]).SARS-CoV-2 syndrome ikaze yubuhumekero coronavirus 2, RT-LAMP revers transcription loop medrated isothermal amplification, abafatanyabikorwa ba tekinoroji ya FTA, NA nucleic aside
Ibice bya magnetiki byuzuye neza nibyiza guhuza umugongo wa fosifate ya acide nucleique.Mugihe cyumunyu mwinshi, amatsinda ya fosifate yuzuye ya acide nucleic arashobora kwishyurwa neza hejuru yubutaka bwa magnetique.Kubwibyo, nanoparticles ya magnetiki ifite ubuso butagaragara hamwe nubucucike bwinshi bwamatsinda ya amino byakozwe kugirango hakurwe aside nucleique.Nyuma yo gutandukana kwa magneti no guhagarika, nanoparticles ya magnetique hamwe na ADN bigizwe na ADN birashobora gukoreshwa muburyo butaziguye muri PCR, bikuraho ibikenewe byo kweza no gufata igihe kandi bitwara igihe [35].Magnetic nanoparticles yashizwe hamwe nitsinda rito rya carboxyl nayo yakoreshejwe mugutandukanya acide nucleic acide yanditswe hejuru yubuso bwa polyethylene glycol hamwe na sodium ya chloride ya sodium [36].Hamwe naya masaro yahinduwe ya magnetiki, gukuramo ADN birahujwe no gukwirakwizwa nyuma.Dignan n'abandi.39Byongeye kandi, ubwuzuzanye bwa ADN yitaruye hamwe na LAMP, uburyo bukwiranye no gusesengura ingingo-yo-kwita kuri acide nucleic aside, irerekana kandi ibikoresho bike bisabwa kandi bikwiranye no gupima amabara (Ishusho 2b).
Uburyo bw'isaro rya Magnetique butanga amahirwe yo gukuramo mu buryo bwikora, bumwe muri bwo bukaba buri mu bucuruzi bwa acide nucleic acide yubucuruzi [KingFisher;ThermoFisher (Waltham, MA, USA), QIAcube® HT;CapitalBio (Beijing, Ubushinwa) na Biomek®;Beckman (Miami, Amerika).), Florida, Amerika)].Ibyiza byo guhuza amasaro ya magnetiki na microfluidics birashobora gukoreshwa mugukuramo neza acide nucleic acide, bishobora guteza imbere iterambere ryisuzumabumenyi;icyakora, guhuza amasaro ya magneti na microfluidics biracyashingira cyane kuri sisitemu igoye yo kugenzura neza uburyo bwo gukoresha neza amasaro ya magneti, ibyo bikaba bisobanura ko ibicuruzwa by’ubucuruzi ari byinshi kandi bihenze, ibyo bikaba bigabanya ikoreshwa ry’amasaro ya magneti muri POCT.
Ibikoresho byinshi nka filozofiya ya nitrocellulose yahinduwe, Ikarita ya Finders Technology Associates (FTA), amakarita ya filteri ya polyethersulfone, hamwe nibikoresho bya glycan nabyo byakoreshejwe mugushakisha aside nucleique [40, 41, 42, 43, 44].Ibikoresho byinshi bya fibrous nkimpapuro za fibrous byakoreshejwe bwa mbere mu gutandukanya ADN mu guhuza umubiri wa molekile ya ADN imaze igihe kirekire hamwe na fibre.Uturemangingo duto tuganisha ku kugabanuka gukomeye kwa molekile ya ADN, bigira ingaruka nziza mu gukuramo ADN.Bitewe nubunini butandukanye bwimpapuro za fibrous, uburyo bwo kuyikuramo ntibushobora guhaza ibyifuzo bya ADN byiyongera [45, 46].Ikarita ya FTA ni impapuro zungurura ubucuruzi zikoreshwa mubijyanye nubuvuzi bwubucamanza kandi zikoreshwa cyane mubindi bice byo gusuzuma molekile.Binyuze mu gukoresha akayunguruzo ka selile yatewe mu miti itandukanye kugira ngo yandike uturemangingo twa selile, ADN yarekuwe irindwa kwangirika kugeza ku myaka 2.Vuba aha, impapuro za selile zatewe mu nda zakozwe kugira ngo hamenyekane molekile zitandukanye ziterwa na virusi nka SARS-CoV-2, leishmaniyasi, na malariya [47,48,49].Virusi itera sida muri plasma yitaruye iraterwa mu buryo butaziguye, kandi aside nucleique ya virusi ikungahaye muri membrane ya FTA® yubatswe mu cyerekezo, ibyo bigatuma umusaruro wa aside nucleique ukora neza [50] (Ishusho 2c).Ikibazo nyamukuru cyo kumenya aside nucleic ukoresheje amakarita ya FTA nuko imiti nka guanidine na isopropanol ibuza reaction ya amplification nyuma.Kugira ngo iki kibazo gikemuke, twateje imbere impapuro zo mu bwoko bwa Fusion 5 chitosan zahinduwe, zihuza ibyiza byo guhuza umubiri kwa molekile ya ADN hamwe nimpapuro zungurura fibrous, hamwe na electrostatike adsorption ya ADN kuri chitosan yahinduwe kugirango tugere kuri acide nucleic acide cyane. ..gushungura fibre [51] (Ishusho 2d).Mu buryo nk'ubwo, Zhu n'abandi.[52] yerekanye uburyo bwa chitosan bwahinduwe bwa PCR bushingiye kuri sisitemu ya microfluidic ya capillary yo kwigunga no kumenya virusi ya Zika RNA.Nucleic acide irashobora kwamamazwa / gutondekwa muburyo bwa lysate / PCR ivanze, ukurikije imitungo ya chitosan.kuri no kuzimya ”, yitabira pH.
Nkuko byavuzwe haruguru, izi ngamba zihuza ibyiza byibikoresho bitandukanye byingenzi kandi byongera imikorere yo gukuramo aside nucleic muri microfluidics.Mubikorwa bifatika, gukoresha ibyo bikoresho kubwinshi ntabwo ari ubukungu, kandi kuvura neza hejuru cyangwa guhindura ubuso bwibikoresho bisanzwe hamwe nibi bikoresho nabyo birashobora kubungabunga imikorere yabyo.Kubwibyo, byizerwa ko ishyirwa mubikorwa ryizi ngamba nyuma yubushakashatsi bwikigereranyo rishobora kugabanya ibiciro.
Kwipimisha aside nucleique kuri platifomu ya microfluidic ikunze gukoresha urugero ruto ruto (<100 µl), bityo rero bisaba ko hongerwaho aside nucleic acide igenewe hamwe na probe yihariye yo guhinduranya ikimenyetso cyorohereza kumenya neza (optique, amashanyarazi, na magnetique) [53, 54]. Kwipimisha aside nucleique kuri platifomu ya microfluidic ikunze gukoresha urugero ruto ruto (<100 µl), bityo rero bisaba ko hongerwaho aside nucleic acide igenewe hamwe na probe yihariye yo guhinduranya ikimenyetso cyorohereza kumenya neza (optique, amashanyarazi, na magnetique) [53, 54]. При тестировании нуклеиновых кислот на микрожидкостных платформах часто используются небольшие объемы образцов (< 100 мкл), поэтому требуется амплификация целевых нуклеиновых кислот с помощью специальных зондов для преобразования в сигнал, удобный для последующего обнаружения (оптического, электрического и магнитного) [53, 54]. Iyo ugerageza acide nucleic kuri platifomu ya microfluidic, hakoreshwa urugero ruto ruto (<100 µL), bityo rero amplification ya acide nucleic acide hamwe na probe idasanzwe irasabwa kuyihindura mubimenyetso byoroshye kugirango bimenyekane nyuma (optique, amashanyarazi, na magnetique) [53, 54].53 54 控 平台 上 的 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 ]。微 流 控 平台 上 的 核酸 100 <100 µl） ， 因此 需要 特定 扩增 扩增 53 53 53 53 ]。 Обнаружение нуклеиновых кислот на микрожидкостных платформах обычно использует небольшие объемы образцов (<100 мкл), что требует амплификации целевых нуклеиновых кислот с помощью специальных зондов для преобразования в сигналы для последующего обнаружения (оптического, электрического и магнитного) [53, 54]]. Kumenya acide nucleic kuri platifomu ya microfluidic mubusanzwe ikoresha urugero ruto rw'icyitegererezo (<100 μl), bisaba ko hongerwaho aside nucleic acide hamwe na probe zidasanzwe kugirango zihindurwe mubimenyetso kugirango bimenyekane nyuma (optique, amashanyarazi, na magnetique) [53, 54]] .Kwiyongera kwa acide nucleique muri microfluidics birashobora kandi kwihutisha ibisubizo, kugabanya imipaka yo gutahura, kugabanya ibyitegererezo, no kunoza neza kumenya neza [55, 56].Mu myaka yashize, hamwe no kumenya vuba na bwangu kumenya, uburyo butandukanye bwo kongera aside nucleic aside bwakoreshejwe muri microfluidics, harimo PCR hamwe na reaction ya isothermal amplification reaction.Iki gice kizavuga muri make uburyo bwo kumenya aside nucleique ishingiye kuri sisitemu ya microfluidic.
PCR niyigana uburyo bwo kwigana ADN yibinyabuzima, theorie yayo isobanurwa muburyo burambuye ahandi kandi ntabwo izaganirwaho hano.PCR irashobora kwongerera umubare muto cyane intego ya ADN / RNA ku gipimo cyerekana, bigatuma PCR igikoresho gikomeye cyo kumenya vuba acide nucleic.Mu myaka ya vuba aha, ibikoresho byinshi byimukanwa bya microfluidic bifite ibikoresho bya sisitemu yo gusiganwa ku magare ya PCR byashyizweho kugira ngo bisuzume ibikenewe byo kwisuzumisha [57, 58].Kuri chip PCR irashobora kugabanywamo amoko ane (asanzwe, akomeza gutemba, guhinduranya ahantu hamwe, hamwe na PCR ya convective PCR) ukurikije uburyo butandukanye bwo kugenzura ubushyuhe [59].Kurugero, Gee n'abandi.[60] yashyizeho uburyo butaziguye bwo kwandukura umubare wa PCR (RT-qPCR) ku mbuga zabo bwite za microfluidic kugira ngo hamenyekane Multlex yo kumenya SARS-CoV-2, virusi ya grippe A na B mu ngero zo mu muhogo (Ishusho 3a).Parike n'abandi.[61] yubatse chip yoroheje yisesengura yindwara ihuza firime yoroheje PCR, electrode, hamwe na polydimethylsiloxane ikoreshwa nintoki ya microfluidic module.Ariko, imirimo yombi ikubiyemo amakosa asanzwe ya PCR isanzwe.PCR isaba gusiganwa ku magare yumuriro, igabanya ibikoresho bya miniaturizasi kandi bigabanya igihe cyo kugerageza.
Iterambere ryikomeza rishingiye kuri microfluidic hamwe na PCR ihinduranya umwanya ni ngombwa kugirango iki kibazo gikemuke.Ukoresheje umuyoboro muremure winzoka cyangwa umuyoboro mugufi ugororotse, guhora utemba PCR irashobora gutanga amplifisation yihuse ikwirakwiza reagent muri zone eshatu zishyushye hamwe na pompe idafite chip.Iki gikorwa kirinda neza icyiciro cyinzibacyuho hagati yubushyuhe butandukanye bityo bigabanya cyane igihe cyo kwipimisha [62] (Ishusho 3b).Mu bundi bushakashatsi bwakozwe na Jung n'abandi.[63] yatanze icyifuzo gishya cyo gusesengura PCR ikomatanya ibiranga PCR ihamye kandi itemba PCR kuri ultrafast na multiplex revers transcription PCR (Ishusho 3c).Kwiyongera kwa acide nucleic, microchip ya PCR izunguruka binyuze mubice bitatu bishyushya mubushyuhe butandukanye: 1. Guhagarika ibice 94 ° C, 2. Guhagarika Annealing kuri 58 ° C, 3. Inzira yo kwaguka kuri 72 ° C.
Gukoresha PCR muri microfluidics.Igishushanyo cyerekana dirRT-qPCR kuri microfluidic platform (yakuwe kuri [60]).b Igishushanyo mbonera cyerekana microarray ikomeza PCR ishingiye kumuyoboro winzoka (yakuwe kuri [62]).c Igishushanyo mbonera cyerekana isesengura rya genoside ya PCR, igizwe na microchip, ibyuma bitatu bishyushya hamwe na moteri ikandagira (yakuwe kuri [63]).d Igishushanyo cya thermoconvection PCR hamwe na centrifugation no gushiraho (byahinduwe kuva [64]).DirRT-qPCR, itaziguye igereranya reaction ya polymerase urunigi
Ukoresheje capillaries hamwe nudusimba cyangwa isahani yoroheje, convection PCR irashobora kwongerera vuba acide nucleic acide kubisanzwe byubushyuhe bwumuriro bidakenewe pompe yo hanze.Kurugero, urubuga rwa cyclic olefin polymer microfluidic rwatejwe imbere murwego rwo guhimba ruzunguruka rukoresha igare ryumuriro hamwe na centrifugation muri microcran ya PCR [64] (Ishusho 3d).Igisubizo cyibisubizo biterwa na convection yumuriro, ikomeza guhanahana ubushyuhe bwo hejuru kandi buke muri microchannel ifite imiterere yumwaka.Igikorwa cyose cyo kwongera imbaraga kirashobora kurangira muminota 10 hamwe na 70.5 pg / umuyoboro.
Nkuko byari byitezwe, PCR yihuta nigikoresho gikomeye cyo guhuza byuzuye-ibisubizo bya molekuline yo gusuzuma no gusesengura ibintu byinshi.PCR yihuta igabanya cyane igihe gisabwa kugirango tumenye SARS-CoV-2, igira uruhare mu kurwanya neza icyorezo cya COVID-19.
PCR isaba igare ryumuriro utoroshye ridakwiriye POCT.Vuba aha, tekinoroji yo kongera ingufu za isothermal yakoreshejwe kuri microfluidics, harimo ariko ntabwo igarukira kuri LAMP, amplification ya recombinase polymerase (RPA), hamwe na amplification ishingiye kuri acide nucleique ikurikirana [65,66,67,68].Hamwe nubu buhanga, acide nucleic yongerewe ubushyuhe buri gihe, byorohereza kurema igiciro gito, cyoroshye cyane cyimodoka ya POCT yifashishwa mugupima molekile.
Microfluidics ishingiye cyane kuri LAMP ivuga ko ituma abantu benshi bamenya indwara zanduza [42, 69, 70, 71].Hamwe na sisitemu ya microfluidic ya centrifugal, LAMP irashobora kurushaho koroshya uburyo bwo kumenya aside nucleique [69, 72, 73, 74, 75].SlipChip izunguruka-yerekana uburyo bwakozwe kugirango hamenyekane bagiteri nyinshi zibangikanye hakoreshejwe LAMP [76] (Ishusho 4a).Iyo ukoresheje LAMP nziza cyane mubisubizo, igipimo cya fluorescence cyerekana urusaku rwikubye inshuro 5, kandi igipimo cyo gutahura cyageze kuri kopi 7.2 / μl za ADN genomic. Byongeye kandi, kubaho kwa bagiteri eshanu zisanzwe ziterwa na bagiteri, zirimo Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis na Vibrio parahaemolyticus, zerekanwe hashingiwe ku buryo muri <60 min. Byongeye kandi, kubaho kwa bagiteri eshanu zisanzwe ziterwa na bagiteri, zirimo Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis na Vibrio parahaemolyticus, zerekanwe hashingiwe ku buryo muri <60 min.Byongeye kandi, kuba hari bagiteri eshanu zitera indwara zifata igifu, zirimo Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis na Vibrio parahaemolyticus, byagaragaye hakoreshejwe ubu buryo mu minota itarenze 60.60 ， 基于 该 方法 60 <60 分钟 内 可视 化 了 五种 常见 消化道 细菌 芽孢 的 、 ， 、 芽孢 杆菌60 ， 基于 该 方法 60 <60 分钟 内 视 化 了 五 种 常见 细菌 芽孢 杆菌 、 ， 肠杆菌弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 HIPByongeye kandi, kuba hari bagiteri eshanu zisanzwe ziterwa na gastrointestinal, harimo nka Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvius, na Vibrio parahaemolyticus, byagaragaye hakoreshejwe ubu buryo mu minota itarenze 60.
Ibyiza bya LAMP muri microfluidics harimo, mubindi, igisubizo cyihuse no gutahura miniaturized.Ariko, kubera ubushyuhe bwa reaction (hafi 70 ° C), byanze bikunze aerosole ikorwa mugihe cya LAMP, bikavamo igipimo cyiza cyibinyoma.Suzuma umwihariko, igishushanyo mbonera, hamwe no kugenzura ubushyuhe nabyo bigomba kuba byiza kuri LAMP.Mubyongeyeho, ibishushanyo mbonera bishyira mubikorwa intego nyinshi kuri chip imwe bifite agaciro gakomeye kandi bigomba gutezwa imbere.Mubyongeyeho, LAMP ikwiranye nintego nyinshi zo gutahura zinjijwe muri chip imwe, ifite akamaro kanini, ariko haracyari ibyumba byinshi byiterambere.
Igipimo kinini cyibinyoma cya LAMP kirashobora kugabanuka igice hamwe na RPA, kuko ubushyuhe buke ugereranije (~ 37 ° C) bivamo ibibazo bike byo guhumeka [77].Muri sisitemu ya RPA, primers ebyiri zinyuranye zitangiza synthesis ya ADN ihuza recombinase kandi amplification irashobora kurangira muminota 10 [78,79,80,81].Kubwibyo, inzira yose ya RPA irihuta cyane kuruta PCR cyangwa LAMP.Mu myaka yashize, tekinoroji ya microfluidic yerekanwe kurushaho kunoza umuvuduko nukuri kwa RPA [82.83.84].Kurugero, Liu n'abandi.[85] yateje imbere microfluidic ihuriweho na polymerase recombinase amplification assay kugirango igaragaze byihuse kandi byoroshye SARS-CoV-2 ihuza inyandiko-mvugo ya RPA (RT-RPA) hamwe na sisitemu yo kwipimisha impande zose.muri sisitemu imwe ya microfluidic.Igicapo 4b).Umupaka wo gutahura ni kopi 1 / µl cyangwa 30 kopi / icyitegererezo, kandi gutahura birashobora kurangira muminota 30.Kong n'abandi.bakoze ibikoresho byenda kwambara microfluidic.[86] yakoresheje ubushyuhe bwumubiri hamwe na terefone igendanwa ishingiye kuri fluorescence yo gutahura kugirango yihute kandi itahure ADN ya VIH-1 ikoresheje RPA (Ishusho 4c).Ikizamini gishobora kwambarwa RPA cyerekana kopi 100 / mL yikurikiranya ryateganijwe mugihe cyiminota 24, byerekana imbaraga nyinshi zo gusuzuma vuba vuba abana banduye virusi itera SIDA-1 ahantu hadafite amikoro make.
Kwiyongera kwa Isothermal muri point-of-care (POCT).Gutezimbere no kubyara spin na reaction SlipChip.Nyuma yo gusudira plasma, chip yo hejuru no hepfo yakusanyirijwe hamwe hamwe nimbuto kugirango ikore chip ya nyuma (yakuwe kuri [76]).b Igishushanyo cya MI-IF-RPA sisitemu yo kumenya COVID-19 (yakuwe kuri [85]).c Igishushanyo cyikizamini cya RPA cyambara kugirango hamenyekane vuba ADN ya VIH-1 (yakuwe kuri [86]).SE Salmonella enterica, VF Vibrio fluvius, VP Vibrio parahaemolyticus, BC Bacillus cereus, EC Escherichia coli, FAM carboxyfluorescein, virusi ya immunodeficiency virusi itera SIDA, RPA recombinase polymerase amplification, LED itanga diode, MI-IF-RPA Microfluidics Integrated L Amplification
Microfluidic ishingiye kuri RPA iratera imbere byihuse, ariko, ikiguzi cyo guhimba chip no gukoresha reaction ni kinini cyane kandi kigomba kugabanuka kugirango hongerwe kuboneka iryo koranabuhanga.Byongeye kandi, ibyiyumvo byinshi bya RPA birashobora kugira ingaruka ku kongera ibicuruzwa bidasanzwe, cyane cyane iyo hari umwanda.Izi mbogamizi zishobora kugira ingaruka kumikoreshereze ya RPA muri sisitemu ya microfluidic kandi bikwiye kurushaho kunozwa.Byateguwe neza na primers hamwe nubushakashatsi ku ntego zitandukanye nazo zirakenewe kugirango tunonosore ingamba za microfluidic zishingiye kuri RPA muri POCT.
Cas13 na Cas12a bafite ubushobozi bwo gukuramo acide nucleic acide bityo birashobora gutezwa imbere nkibikoresho byo kumenya no gusuzuma.Cas13 na Cas12a zikora mugihe cyo guhuza ADN cyangwa RNA.Iyo poroteyine imaze gukora, itangira gukuramo andi acide nucleic iri hafi, hanyuma ikayobora RNAs yibasira aside nucleic acide yihariye ishobora gukuramo florescent yazimye ikarekura fluorescence.Ukurikije iki gitekerezo, Kellner n'abandi.][88] yashyizeho ubundi buryo bushingiye kuri Cas12a [CRISPR Trans Reporter yibasira ADN endonuclease (DTECR)].
Mu myaka yashize, hagaragaye uburyo butandukanye bwo kumenya aside nucleique ishingiye kuri CRISPR [89, 90].Uburyo busanzwe bwa CRISPR akenshi butwara igihe kandi bukora cyane kubera inzira nyinshi zirimo gukuramo aside nucleic, amplification no kumenya CRISPR.Guhura n'amazi mu kirere bishobora kongera amahirwe y'ibisubizo byiza.Urebye ibyavuzwe haruguru, sisitemu ishingiye kuri CRISPR ikeneye cyane optimizas.
Porogaramu ya microfluidic igenzurwa na pneumatike ishobora gukora isesengura 24 murwego rumwe yateguwe kubikorwa bya CRISPR-Cas12a na CRISPR-Cas13a yo gushakisha [91].Sisitemu ifite ibikoresho byo gutahura fluorescence irenga aside nucleic aside kandi igahita imenya ADN femtomolar na RNA.Chen n'abandi.[92] guhuza imbaraga za recombinase hamwe na sisitemu ya CRISPR-Cas12a muri microfluidics ya centrifugal (Ishusho 5a).Aka kazi katsinze ingorane zo guhuza izi nzira zombi kuko Cas12a ishobora gusya ADN yintumwa kandi ikabuza inzira yo kongera imbaraga.Byongeye kandi, Chen n'abandi.[92] wongeyeho kubanza kubika reaction reagent muri microfluidic igenzura kugirango ihite irangiza inzira yose.Mu kindi gikorwa, Silva n'abandi.[93] yashyizeho uburyo bwo kwisuzumisha nta CRISPR / Cas12a yongerewe imbaraga na terefone yo kumenya SARS-CoV-2 (Ishusho 5b).Ubu bushakashatsi, buzwi nka sisitemu ya terefone igendanwa ishingiye kuri amplification idafite sisitemu, ikubiyemo enzyme ya CRISPR / Cas ishingiye kuri terefone ishingiye ku iyerekwa rya terefone ryerekana ibimenyetso bya catalase byakozwe na bubble mu miyoboro ya microfluidic.Kumenya neza kopi ziri munsi ya 50 / µl ya acide nucleic nta pre-amplification, inzira yose kuva inshinge zicyitegererezo kugeza gusoma ibimenyetso bifata iminota 71 gusa.
Nucleic aside yo kumenya uburyo bushingiye kuri CRISPR.PIFT ya Centrifugal yo kwisuzumisha ya molekuline ishingiye kuri CRISPR (yakuwe kuri [92]).b Gutezimbere ikizamini cya CASCADE kubisesengura bishingiye kuri terefone ya SARS-CoV-2 (yakuwe kuri [93]).RAA recombinase amplification, PAM yegeranye na protospacer motif, CRISPR yakusanyije palindromic ngufi isubiramo mugihe gisanzwe, sisitemu ya CASCADE idafite terefone igendanwa hamwe na enzymes zishingiye kuri CRISPR / CAS, 1-Ethyl-3- [3-dimethylaminopropyl] karbodiimide hydrochloride EDC
Nintambwe yanyuma mugushakisha aside nucleic, kumenya ibimenyetso byerekana neza ibisubizo byo kwisuzumisha kandi nikintu gikomeye mugutezimbere POCT ikora neza, yunvikana, kandi nyayo.Ibimenyetso birashobora gusomwa ukoresheje uburyo butandukanye nka fluorescent, electrochemic, colorimetric na magnetique.Muri iki gice, turasobanura impamvu zifatika kuri buri buryo kandi tugereranya gupima molekuline yindwara zandura muri microfluidics.
Ingamba zishingiye kuri Fluorescence zikoreshwa cyane mugupima POCT gusuzuma indwara zandura kubera ibyiza byazo byo kumva neza, kugiciro gito, koroshya imikorere, no gusesengura ingingo-yitaweho [94, 95].Izi ngamba zikoresha fluorophores yanditseho amarangi ya fluorescent hamwe na nanomateriali kugirango habeho ikimenyetso kiboneka (kuzamura fluorescence cyangwa kuzimya).Ubu bushakashatsi bwerekana ko ingamba zishingiye kuri fluorescence zishobora kugabanywa mu buryo butaziguye ibimenyetso bya fluorescent, ibimenyetso-byerekana, hamwe no kumenya ibimenyetso bya fluorescent [96].Ikimenyetso cya fluorescent itaziguye ikoresha ibirango byihariye bya fluorescente kugirango yandike ligande yihariye itanga urugero rwihariye rwa fluorescence mugihe ihujwe nintego.Kumenyekanisha ibimenyetso bishingiye kuri fluorescence, ubwiza bwikimenyetso cya fluorescent bifitanye isano neza nubunini bwinyungu.Imbaraga za Fluorescence ntizihagije mugihe habuze intego kandi ziramenyekana mugihe intego ihagije ihari.Ibinyuranye, ubukana bwa fluorescence bwagaragajwe na "signal-off" fluorescence ihwanye neza nubunini bwintego, ubanza igera ku giciro kinini kandi igenda igabanuka buhoro buhoro uko intego yagutse.Kurugero, ukoresheje CRISPR-Cas13a intego-iterwa na trans-cleavage uburyo, Tian nibindi.[97] yashyizeho ingamba zo kumenyekanisha udushya kugira ngo tumenye RNAs zinyura mu buryo butaziguye (Ishusho 6a).Iyo uhujwe no kuzuza intego za RNAs, uruganda rwa CRISPR-Cas13-RNA rushobora gukora, bigatuma habaho gutandukana hagati yumunyamakuru RNAs udasanzwe.Florescent yanditseho umunyamakuru [fluorophore (F)] azimya na quencher (Q) idahwitse na fluoresces iyo ikozwe nikigo gikora.
Ibyiza byo kumenya amashanyarazi ni umuvuduko mwinshi wo gutahura, umusaruro woroshye, igiciro gito, byoroshye gutwara no kugenzura byikora.Nuburyo bukomeye bwo gusesengura kubikorwa bya POCT.Ukurikije graphene umurima-ngaruka transistors Gao nabandi.[98] yashyizeho nanobiosensor kugirango igaragaze antigene yindwara ya Lyme ikomoka kuri bagiteri ya Borrelia burgdorferi ifite igipimo cya 2 pg / mL (Ishusho 6b).
Ibara ryibara ryakoreshejwe mubisabwa POCT, byungukirwa nibyiza byo gutwara, igiciro gito, koroshya imyiteguro, no gusoma neza.Gutahura amabara birashobora gukoresha okiside ya peroxidase cyangwa peroxidase imeze nka nanomateriali, kwegeranya nanomateriali, hamwe no kongeramo amarangi yerekana ibimenyetso kugirango uhindure amakuru ajyanye no kuba acide nucleic acide ihinduka mumabara agaragara [99, 100, 101].Ikigaragara ni uko nanoparticles ya zahabu ikoreshwa cyane mugutezimbere ingamba zamabara, kandi bitewe nubushobozi bwabo bwo guhindura amabara yihuse kandi akomeye, hagenda hagaragara ubushake bwo guteza imbere urubuga rwa POCT rwerekana uburyo bwo gusuzuma indwara zanduye [102].Hamwe na microfluidic igizwe na microfluidic igizwe na 103, indwara ziterwa na virusi ziterwa n'amata yanduye zirashobora guhita zimenyekana kurwego rwa selile 10 za bagiteri, kandi ibisubizo birashobora gusomwa mumashusho muminota 65 (Ishusho 6c).
Tekinike ya magnetiki irashobora kumenya neza analyite ukoresheje ibikoresho bya magneti, kandi hari inyungu zikomeye mubikorwa bya POCT mumyaka mirongo ishize.Tekiniki ya magnetiki tekinike ifite ibyiza byihariye nkibikoresho bya magneti bihenze kuruta ibikoresho bihenze bya optique.Ariko, gukoresha umurima wa magneti bitezimbere imikorere yo gutahura kandi bigabanya igihe cyo gutegura icyitegererezo [104].Byongeye kandi, ibisubizo byubushakashatsi bwa magneti byerekana umwihariko, ibyiyumvo, hamwe n’ikigereranyo kinini cyerekana urusaku bitewe n’ikimenyetso kidasanzwe cya magnetiki cyerekana ibimenyetso by’ibinyabuzima [105].Sharma n'abandi.yahujije magnetiki tunnel ihuza biosensor muburyo bworoshye bwa microchip.[106] kugirango hamenyekane ibintu byinshi bitera indwara (Ishusho 6d).Biosensor itahura neza acanike nucleic acide itandukanijwe na virusi.
Uburyo busanzwe bwo kumenya ibimenyetso.Igitekerezo cyo kumenya hyperlocalized detection ya Cas13a (yakuwe muri [97]).b Graphene nanobiosensor FET ifatanije na Lyme GroES scFv (yakuwe muri [98]).c Ibimenyetso byerekana amabara menshi yo gutahura indwara ziterwa na mikorobe ya microfluidic chip: No 1 na No 3 icyitegererezo hamwe na virusi zanduye, na No 2, No 4 na No 5 ntangarugero zidafite virusi (zahinduwe kuva [103]) .d Biosensor ishingiye ku ihuriro rya rukuruzi ya rukuruzi, harimo urubuga, rwubatswe rwuzuza ibyuma byongera imbaraga, ishami rishinzwe kugenzura, hamwe n’amashanyarazi yo gutanga ibimenyetso / kugura (byahinduwe kuva [106]).GFET Graphene FET, Escherichia coli, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus, Vibrio parahaemolyticus, Listeria monocytogene, PC PC, PDMS Dimethicone, PMMA polymethyl methacrylate
Nubwo ibintu byiza biranga uburyo bwo gutahura hejuru, baracyafite ibibi.Ubu buryo bugereranijwe (imbonerahamwe 1), harimo porogaramu zimwe zifite ibisobanuro (ibyiza n'ibibi).
Hamwe niterambere rya microfluidics, sisitemu ya microelectromechanique, nanotehnologiya nibikoresho bya siyansi, ikoreshwa rya microfluidic chip kugirango hamenyekane indwara zandura rihora ritera imbere [55,96,107,108].Gukoresha neza ibikoresho bito na fluide bigira uruhare mugusuzuma neza no gukoresha neza.Kubwibyo, kugirango turusheho gutera imbere, hashyizweho ingufu zo kunoza no kuzamura chip, bivamo microfluidic chip zitandukanye hamwe nimirimo itandukanye.Hano turamenyekanisha muri make ubwoko butandukanye bwa microfluidic kandi tugereranya ibiranga (ibyiza n'ibibi).Mubyongeyeho, ingero nyinshi zavuzwe hepfo zibanda cyane cyane kurwanya SARS-CoV-2.
LOCCs ni sisitemu yo gusesengura cyane ya miniaturizasi kandi ibikorwa byayo ni miniaturike cyane, ihuriweho, ikora kandi igereranywa no guterwa icyitegererezo no gutegura, kugenzura imigezi no gutahura amazi [109, 110].Amazi akoreshwa binyuze muri geometrie yateguwe neza no gukorana kwingaruka nyinshi zumubiri nka gradients yumuvuduko, ibikorwa bya capillary, electrodynamic, imirima ya magnetique hamwe numuraba wa acoustic [111].LOCC yerekana ibyiza bihebuje mugusuzuma byinshi-byinjira no gutahura byinshi, hamwe nisesengura ryihuse, ingano ntoya, gukoresha ingufu nke, hamwe nubuyobozi bukora neza kandi bukora neza;icyakora, ibikoresho bya LOCC biroroshye cyane, kandi gukora, gupakira, no guhuza.Ariko, kugwiza no gukoresha byongeye guhura ningorane zikomeye [96].Ugereranije nandi ma platform, LOCC ifite ibyiza byihariye mubijyanye nuburyo butandukanye bwo gukoresha no guhuza ikoranabuhanga ryiza, ariko ibibi byayo nabyo biragaragara, aribyo bigoye cyane kandi bidasubirwaho.Biterwa na pompe zo hanze, akenshi usanga ari nini kandi zihenze, bikomeza kugabanya imikoreshereze yabo muri POCT.
Mugihe cya COVID-19, LOCC yitabiriwe cyane.Mugihe kimwe, hariho chip nyinshi nshya zihuza tekinoroji nyinshi.Kurugero, terefone zigendanwa zikoreshwa cyane nkibikoresho byisesengura byoroshye kandi bifite amahirwe menshi yo guhuza LOCC.Izuba n'abandi.[21] yahimbye chip ya microfluidic yemerera kugwiza aside nucleic yihariye ikurikirana ya virusi eshanu, harimo na SARS-CoV-2, ikoresheje LAMP ikanabisesengura ukoresheje terefone mugihe cyisaha 1 nyuma yuko reaction irangiye.Urundi rugero, Sundah n'abandi.112 CATCH irahuzwa na LOCC igendanwa kandi igera kubikorwa byiza (hafi 8 kopi ya RNA / μl; <1 h mubushyuhe bwicyumba) [112]. CATCH irahuzwa na LOCC igendanwa kandi igera kubikorwa byiza (hafi 8 kopi ya RNA / μl; <1 h mubushyuhe bwicyumba) [112]. CATCH совместим с портныным LOCC и обеспечивает превосходную про проититнность (примно 8 копий РНК / мкл; <1 ч при комнатной гее) [112]. CATCH irashobora guhuzwa na LOCC igendanwa kandi itanga ibicuruzwa byiza (hafi 8 RNA kopi / µl; <1 h mubushyuhe bwicyumba) [112]. FATA 与 便携式 LOCC 兼容 并 具有 卓越 的 性能 （大约 8 RNA 拷贝 / μl ； 室温 下 <1 小时） [112]。 FATA 与 便携式 LOCC 兼容 并 具有 卓越 的 性能 （大约 8 RNA 拷贝 / μl ； 室温 下 <1 小时） [112]。 CATCH совместим с портныными LOCC и обладает превосходной про проитительносту (примерно 8 копий РНК / мкл; <1 часа при комнатной баре) [112]. CATCH ihujwe na LOCCs igendanwa kandi ifite imikorere myiza (hafi 8 kopi ya RNA / µl; <isaha 1 mubushyuhe bwicyumba) [112].Mubyongeyeho, ibikoresho bya LOCC byo gusuzuma molekuline nabyo bikoresha imbaraga zimwe zo gutwara nka vacuum, kurambura, hamwe namashanyarazi.Kang n'abandi.]Li n'abandi.[114] nyuma yaje gukora chip-microfluidic chip irambuye ifasha gusuzuma COVID-19.Ihuriro rikoresha sisitemu yo kongera ingufu za RT-LAMP kugirango hamenyekane niba icyitegererezo ari cyiza cyangwa cyiza.Nyuma, Ramachandran n'abandi.[115] yageze kumashanyarazi akwiye akoresheje isotachophoresis (ITP), tekinike yatoranijwe yibanda kuri microfluidics.Hamwe na ITP, intego ya RNA kuva nasopharyngeal swab ntangarugero irashobora guhita isukurwa.Hanyuma Ramachandran n'abandi.[115] Gukomatanya uku kweza kwa ITP hamwe na ITP yongerewe imbaraga na LAMP na CRISPR ivuga ko byagaragaye SARS-CoV-2 muri nasopharyngeal swab yumuntu hamwe nubuvuzi bwa clinique muminota igera kuri 35.Byongeye kandi, ibitekerezo bishya bigenda bigaragara.Jadhav n'abandi.]Membrane-ikora-yubatswe muyungurura microchannel irashobora gukoreshwa.Igikoresho cyamamaza virusi ziva mumazi atandukanye yumubiri / gusohora nka macandwe, nasofarynx n'amarira.Rero, titer ya virusi ni myinshi kandi virusi irashobora kumenyekana neza numukono wa Raman.
UMUYOBOZI ni microfluidic ya centrifugal aho inzira zose zigenzurwa na protocole yumurongo uzunguruka microstructures substrate [110].Igikoresho LOAD kirangwa no gukoresha imbaraga za centrifugal nkimbaraga zikomeye zo gutwara.Amazi nayo akoreshwa na capillary, Euler na Coriolis.Ukoresheje igikoresho cya centrifuge, isesengura rikorwa mugikorwa cyamazi gikomeza kuva mumirasire yimbere kugeza kumwanya winyuma, bikuraho ibikenerwa byongeweho hanze, pompe, moteri, na valve ikora.Muri make, uburyo bumwe bwo kugenzura bworoshya imikorere.Imbaraga zikora kumazi mumurongo umwe wa microfluidic kumurongo umwe uvuye hagati yikigo kiremereye, ibyo bigatuma bishoboka gusubiramo imiterere yumurongo.Rero, ibikoresho LOAD biroroshye kandi byubukungu gushushanya no gukora kuruta ibikoresho bisanzwe bya LOCC, mugihe ibisubizo ahanini byigenga kandi birasa;icyakora, kubera imbaraga nyinshi zubukanishi bwibikoresho bya centrifugal, ibikoresho bya chip biboneka ni bike kandi ingano nto biragoye.ku modoka.Muri icyo gihe, ibikoresho byinshi BIKURIKIRA bigenewe gukoreshwa rimwe gusa, bihenze kubushakashatsi bunini [96, 117, 118, 119].
Mu myaka ya vuba aha, LOAD, ifatwa nk'imwe mu bikoresho bitanga icyizere cya microfluidic, yitabiriwe n'abashakashatsi n'ababikora.Rero, LOAD imaze kwemerwa cyane kandi yakoreshejwe mugupima molekuline yanduye virusi [120, 121, 122, 123, 124], cyane cyane mugihe cya COVID-19.Kurugero, mu mpera za 2020, Ji n'abandi.[60] yerekanye uburyo butaziguye RT-qPCR kugirango hamenyekane byihuse kandi byikora mu buryo bwihuse bwo kumenya SARS-CoV-2 hamwe na grippe A na B zanduye mu muhogo.Hanyuma Xiong n'abandi.[74] yerekanye itara rya microfluidic ya LAMP ihuriweho na microfluidic kugirango hamenyekane byihuse, byukuri, kandi icyarimwe icyarimwe cyo kumenya coronavirus zirindwi zubuhumekero zabantu, harimo na SARS-CoV-2, muminota 40.Mu ntangiriro za 2021, de Oliveira n'abandi.73Nyuma, Dignan n'abandi.39Medved n'abandi.53Suarez n'abandi.75Izi ngero zerekana inyungu nini nisezerano RY'UMUYOBOZI mugupima molekuline ya COVID-19.
Mu 1945 Muller na Clegg [125] berekanye bwa mbere imiyoboro ya microfluidic ku mpapuro bakoresheje impapuro zungurura na paraffine.Mu 2007, itsinda rya Whitesides [126] ryashizeho urubuga rwa mbere rukora impapuro zo gupima poroteyine na glucose.Impapuro zahindutse substrate nziza ya microfluidics.Urupapuro rufite ibintu byihariye nka hydrophilicity nuburyo bubi, biocompatibilité nziza, uburemere bworoshye, guhinduka, guhindagurika, igiciro gito, koroshya gukoresha no korohereza.Ibisanzwe µPADs bigizwe na hydrophilique / hydrophobique yubatswe ku mpapuro.Ukurikije imiterere-yuburyo butatu, μPADs irashobora kugabanywamo ibice bibiri (2D) na bitatu-bya (3D) μPADs.2D µPADs ikorwa mugukora imipaka ya hydrophobique kugirango ikore imiyoboro ya microfluidic, mugihe 3D µPAD isanzwe ikorwa mubice byimpapuro za microfluidic 2D, rimwe na rimwe ikabikwa nimpapuro, tekinike zo kunyerera, imiyoboro ifunguye, hamwe no gucapa 3D [96].Amazi yo mu mazi cyangwa ibinyabuzima kuri μPAD agenzurwa cyane cyane nimbaraga za capillary zidafite ingufu zituruka hanze, byorohereza kubikwa mbere ya reagent, gufata ibyitegererezo, no gutahura multiplex.Nyamara, kugenzura neza imigendekere no gutahura ibintu byinshi bibangamiwe numuvuduko udahagije wo gutahura, kumva, no kongera gukoreshwa [96, 127, 128, 129, 130].
Nka porogaramu ya microfluidic idasanzwe, μPAD yazamuwe cyane kandi itezwa imbere kugirango isuzume molekile yindwara zandura nka HCV, VIH, na SARS-CoV-2 [131, 132].Kubihitamo no kumva neza HCV, Tengam nibindi.[133] yakoze biosensor nshyashya ishingiye ku mpapuro za fluorescent ikoresheje probe yihariye ya nucleic aside ishingiye kuri peptide ya pyrrolidinyl.Acide nucleique ihindagurika ku mpapuro za selile ya okisile igice hamwe na alkylation igabanya amatsinda ya amino na aldehyde, kandi gutahura bishingiye kuri fluorescence.Ibi bimenyetso birashobora gusomwa nibikoresho byabugenewe byabugenewe hamwe na kamera ya fluorescent igendanwa hamwe na kamera ya terefone ngendanwa.Nyuma, Lu n'abandi.[134] yateguye impapuro zishingiye kuri electrode yoroheje ishingiye kuri nikel / zahabu nanoparticles / carbone nanotubes / polyvinyl alcool organometallic igizwe no kwandura virusi itera sida hakoreshejwe ADN ivangwa hakoreshejwe methylene ubururu nk'ikimenyetso cya redox ya ADN.Vuba aha, Chowdury n'abandi..
Igeragezwa ryuruhande ruyobora ibiyobora imbaraga za capillary no kugenzura umuvuduko wamazi ukoresheje ubushuhe nibiranga insimburangingo cyangwa microstructures.Ibikoresho bitemba byuruhande bigizwe nicyitegererezo, conjugate, incubator no gutahura, hamwe nudupapuro twinjiza.Molekile ya nucleic acide muri LFA imenya binders zabitswe mbere yabitswe aho zihuza kandi zihuza nkibigo.Mugihe amazi anyuze muri plaque ya incubation na detection, ibigo bifatwa na molekile zifata ziri kumurongo wikizamini no kugenzura, byerekana ibisubizo bishobora gusomwa mumaso.Mubisanzwe, LFA irashobora kurangira muminota 2-15, byihuse kuruta kuvumbura gakondo.Bitewe nuburyo bwihariye, LFA isaba ibikorwa bike kandi ntisaba ibikoresho byinyongera, bigatuma ikoreshwa neza cyane.Nibyoroshye gukora no kugabanya miniaturize, kandi igiciro cyimpapuro zishingiye kubutaka ni gito.Nyamara, ikoreshwa gusa mubisesengura ryujuje ubuziranenge, kandi kumenya umubare biragoye cyane, kandi ubushobozi bwo kugwiza no kwinjiza ni bike cyane, kandi aside imwe gusa ihagije irashobora kuboneka icyarimwe [96.110,127].
Nubwo porogaramu nyinshi za LFA zibanda ku gukingira indwara, gukoresha LFA mugupima molekuline muri chip ya microfluidic nayo irakora kandi irakunzwe [136].Ku bijyanye na virusi ya hepatite B, VIH na SARS-CoV-2 LFA Gong n'abandi.[137] yatanze igitekerezo cyo guhinduranya nanoparticle ya LFA kandi yerekana uburyo butandukanye bwurubuga ruto kandi rwimurwa hifashishijwe uburyo bworoshye bwo kumenya intego nyinshi nka aside HBV nucleic.Mubyongeyeho, Fu n'abandi.]Kugirango byihuse kandi byoroshye kumenya SARS-CoV-2, Liu n'abandi.85
Porogaramu ya microfluidic itandukanye iratandukanye bitewe nubushakashatsi bwihariye, ukoresheje neza ubushobozi nibyiza byurubuga.Hamwe na valve ihendutse, pompe nuyoboro, LOCC ni urubuga rwuzuye rwo gukoresha ibintu bitandukanye no gukorana hamwe nicyumba kinini cyiterambere.Kubwibyo, turizera kandi turasaba ko ubushakashatsi bushya bwakorerwa muri LOCC nkigerageza ryambere kandi ko ibintu byagenda neza.Mubyongeyeho, uburyo bunoze kandi bunoze buteganijwe kuvumburwa no gukoreshwa muri sisitemu.UMUYOBOZI urusha abandi kugenzura neza ibintu biva mu bikoresho bya LOCC kandi byerekana ibyiza byihariye muri drives imwe n'imbaraga za centrifugal bidakenewe drives zo hanze, mugihe ibisubizo bisa bishobora gutandukana kandi bigahuzwa.Rero, mugihe kizaza, UMUYOBOZI uzaba urubuga nyamukuru rwa microfluidic hamwe nibikorwa bike byintoki hamwe nikoranabuhanga rikuze kandi ryikora.PlatformPAD ihuza inyungu za LOCC nibikoresho bishingiye ku mpapuro zihenze, gukoresha rimwe gusa kwisuzumisha.Kubwibyo, iterambere rizaza rigomba kwibanda ku ikoranabuhanga ryoroshye kandi ryashizweho neza.Byongeye kandi, LFA ikwiranye no kumenya amaso yambaye ubusa, isezeranya kugabanya ikoreshwa ryikitegererezo no kwihutisha gutahura.Kugereranya birambuye birambuye bigaragara mu mbonerahamwe ya 2.
Isesengura rya sisitemu rigabanya icyitegererezo muri microreactors nyinshi, buri kimwekimwe kirimo umubare utandukanye wa molekile zigenewe [139, 140].Isuzuma rya Digitale ritanga inyungu zingenzi zo gukora umubare wuzuye mugukora ibihumbi n'ibihumbi bigereranywa na biohimiki icyarimwe kandi kugiti cyayo mugice cya micron aho kuba mugice gikomeza.Ugereranije na microfluidics gakondo, reaction yibice irashobora kugabanya urugero rwicyitegererezo, kongera imikorere yuburyo bwiza, kandi igahuzwa byoroshye nubundi buryo bwo gusesengura bidakenewe imiyoboro, pompe, indangagaciro, hamwe nigishushanyo mbonera [141, 142, 143, 144, 145, 146, 147].Uburyo bubiri bukurikira bukoreshwa muburyo bwa digitale kugirango ugere kubutandukaniro bumwe kandi nyabwo bwibisubizo, harimo reagent hamwe nintangarugero nka selile, acide nucleique, nibindi bice cyangwa molekile: (1) emulisiyo itonyanga ikoresha imiyoboro idahwitse;(2) kugabana umurongo bikorwa nimbogamizi ya geometrike yibikoresho.Muburyo bwa mbere, ibitonyanga birimo reagent hamwe nicyitegererezo muri microchannel birashobora gushirwaho muburyo bworoshye nka co-current, crossflow, flow focus, stade emulisation, emulisation ya microchannel, hamwe na membrane binyuze mumashanyarazi yogosha hamwe na emulisation hamwe no guhindura imiyoboro.kwimenyekanisha [143, 145, 146, 148, 149] cyangwa gukoresha uburyo bukora [150, 151], butangiza ingufu zinyongera binyuze mumashanyarazi, magnetiki, ubushyuhe nubukanishi.Muburyo bwa nyuma, ubunini bwiza bwamazi meza mubyumba bya microfluidic bisangirwa mugukomeza imiterere yubunini bungana, nka micropits hamwe nubuso bwubuso [152,153,154].Ikigaragara ni uko ibitonyanga ari ibice byingenzi bitemba bishobora no kubyara no gukoreshwa kuri electrode ya array ishingiye kuri microfluidics ya digitale (DMF).Electrowetting ya dielectrics ni imwe mu nyigisho za DMF zize neza, kubera ko amashanyarazi ya dielectrics yemerera gukoresha neza ibitonyanga ku giti cye, kugenzura imiterere y’ibimenyetso by’amashanyarazi bitamenyerewe binyura mu mpande zitandukanye [141, 144].Ibikorwa nyamukuru hamwe nigitonyanga muri DMF harimo gutondeka, gucamo ibice, no guhuza [151, 155, 156], bishobora gukoreshwa mubice bitandukanye byisesengura, cyane cyane mugushakisha molekile [157, 158, 159].
Kugaragaza aside nucleic aside ni igisekuru cya gatatu cya tekinoroji yo kwisuzumisha ikurikira PCR isanzwe hamwe numubare nyawo wa PCR (qPCR), ugereranije no kwinjiza ibintu byinshi hamwe na biopsy y'amazi.Mu myaka 20 ishize, acide nucleic acide yateye imbere byihuse mubijyanye no gusuzuma molekuline yanduye virusi yanduye [160, 161, 162].Umubare wuzuye wa acide nucleic aside igaragara utangirana no gupakira ingero hamwe na reagent mubice bitandukanye kugirango barebe ko buri cyiciro gikurikirana gifite amahirwe amwe yo kwinjira muri buri gice.Mubyukuri, buri gice gishobora guhabwa intego nyinshi zikurikirana, cyangwa ntihashobora kubaho sisitemu yigenga ya microreaction.Binyuze muburyo butandukanye bwo kwiyumvisha ibintu byasobanuwe haruguru, ibice bifite intego ya mikorobe bikurikirana bitanga ibimenyetso hejuru yurwego runaka birashobora kugaragazwa nijisho ryonyine cyangwa imashini kandi byanditseho ko ari byiza, mugihe ibindi bice bitanga ibimenyetso munsi yurugero byanditseho ko ari byiza. .ibibi, bituma ibimenyetso kuri buri gice biba.Rero, mukubara umubare wibice byakozwe nigipimo cyibisubizo byiza nyuma yo kubyitwaramo, kopi yumwimerere yikigereranyo cyibizamini irashobora guhuzwa hakoreshejwe formula yo gukwirakwiza Poisson bitabaye ngombwa ko hajyaho umurongo usanzwe, usabwa kubisesengura bisanzwe. nka qPCR.[163] Ugereranije nuburyo bwa gakondo bwo gupima molekuline, gutahura aside nucleic aside ifite urwego rwo hejuru rwo kwikora, umuvuduko mwinshi wo gusesengura no kwiyumvisha ibintu, reagent nkeya, kwanduza gake, no gushushanya no gukora byoroshye.Kubera izo mpamvu, ikoreshwa ryibisubizo bya digitale, cyane cyane uburyo bushingiye ku guta, kubisuzuma bya molekile, guhuza amplification hamwe nubuhanga bwo gusoma ibimenyetso, byizwe neza mugihe gikomeye cya SARS-CoV-2.Kurugero, Yin n'abandi.[164] ikomatanya uburyo bwa digitale nuburyo bwihuse bwa PCR kugirango tumenye genes ya ORF1ab, N, na RNase P muri SARS-CoV-2 muri chip ya microfluidic.Ikigaragara ni uko sisitemu yashoboye kumenya ikimenyetso cyiza mumasegonda 115, yihuta kurusha PCR isanzwe, byerekana akamaro kayo mugushakisha aho bita (Ishusho 7a).Dong n'abandi.[165], Kubiba n'abandi.[157], Chen n'abandi.[166] na Alteri n'abandi.[167] yakoresheje kandi igitonyanga cya digitale PCR (ddPCR) kugirango amenye SARS-CoV-2 muri sisitemu ya microfluidic hamwe nibisubizo bitangaje.Kugirango turusheho kunoza igipimo cyo gutahura, Shen n'abandi.88Ntabwo uburyo bwo gukwirakwiza amashyuza gusa nka PCR bukoreshwa, ariko nuburyo bukoreshwa bwa isothermal amplification bwakoreshejwe kugirango byorohereze imiterere nigisubizo cyihuse.Lu n'abandi.[71] yateje imbere SlipChip yo gusesengura ibitonyanga, ishoboye kubyara ibitonyanga bifite ubunini butandukanye ku ntera ndende mu ntambwe imwe no kugereranya acide nucleic ya SARS-CoV-2 ikoresheje LAMP (Ishusho 7b).Nka tekinoroji yihuta cyane, CRISPR irashobora kandi kugira uruhare runini mugutahura aside nucleic acide binyuze mumashusho yoroshye ya colimetric idakeneye andi moko ya acide nucleic.Ackerman n'abandi.yateje imbere matrix reaction ya multiplex yo gusuzuma acide nucleic.[158] yatahuye virusi 169 zifitanye isano n’abantu, harimo na SARS-CoV-2, mu bitonyanga birimo CRISPR-Cas13 ishingiye kuri nucleic aside yo mu bwoko bwa nucleic acide mu bushakashatsi bwa microwell (Ishusho 7c).Mubyongeyeho, isothermal amplification hamwe na tekinoroji ya CRISPR irashobora gukoreshwa muri sisitemu imwe kugirango ihuze inyungu zombi.Parike n'abandi.169 Ikigereranyo cyigihe., intera nini yagutse hamwe no kumva neza (Ishusho 7d).Ibisobanuro bimwe byizo ngero byatanzwe mu mbonerahamwe ya 3.
Ubusanzwe sisitemu ya digitale yo kumenya aside nucleic.a Byihuta byakazi bya PCR bigizwe nintambwe enye zingenzi: gutegura icyitegererezo, gukwirakwiza imvange ya reaction, uburyo bwo kongera imbaraga, no kugereranya intego (byakuwe kuri [164]).b Igishushanyo cyerekana isesengura ryibitonyanga bya SlipChip kugirango bibe bitonyanga hejuru (byahinduwe kuva [71]).c CARMEN-Cas igishushanyo mbonera cyakazi13 (cyahinduwe kuva [158]).d Incamake yerekana virusi itera imbere igezweho hamwe na CRISPR / Cas mu nkono imwe (yakuwe muri [169]).W / O amazi-y-amavuta, polydimethylsiloxane PDMS, reaction ya PCR polymerase reaction, ikusanyamakuru rya DAQ, inkomoko ya PID igereranya intangiriro, CARMEN ikomatanya matrix reaction yo gusuzuma aside irike ya nucleic, SARS-CoV-2, syndrome ikabije yubuhumekero bukabije, coronavirus 2, RT Kwiyongera kwa transcript transcriptase recombinase polymerase-RPA, S / B ikimenyetso inyuma