News

Nzira dzechinyakare dzekuongorora zvirwere zvekuona zvirwere zvinotapukira dzinoda kushandiswa kwezviridzwa zvebhenji zvisina kukodzera kuongororwa-kutarisira (POCT).Emerging microfluidics yakanyanya miniaturized, otomatiki, uye yakabatanidzwa tekinoroji inova imwe nzira yemaitiro echinyakare yekukurumidza, yakaderera-mutengo, yakarurama pa-saiti diagnostics.Molecular diagnostic nzira dzinoshandiswa zvakanyanya mumidziyo yemicrofluidic senzira dzinoshanda dzekuona pathogen.Ongororo iyi inopfupikisa kufambira mberi kwazvino mu microfluidic-based molecular diagnostics yezvirwere zvinotapukira kubva kune ese ezvidzidzo uye maindasitiri maonero.Kutanga, isu tinotsanangura yakajairwa pa-chip processing ye nucleic acids, kusanganisira sampuli pretreatment, amplification, uye chiratidzo chekuverenga.Hunhu, mabhenefiti uye kuipa kwemhando ina dzemicrofluidic mapuratifomu anobva aenzaniswa.Tevere, isu tichakurukura mashandisirwo edhijitari assays yeabsolute quantification ye nucleic acids.Ese ari maviri echinyakare uye achangoburwa ekutengesa microfluidic-yakavakirwa mamorekuru diagnostic zvishandiso anopfupikiswa sehumbowo hwemamiriro azvino emusika.Chekupedzisira, isu tinopa mazano emangwana eiyo microfluidic kuongororwa kwezvirwere zvinotapukira.
Zvirwere zvinotapukira zvinokonzerwa nezvirwere, kusanganisira mabhakitiriya, mavhairasi, uye utachiona, izvo zvinoparadzirwa pasi rose.Kusiyana nezvimwe zvirwere, utachiona hunokurumidza kutapukirwa uye huchipararira pakati pevanhu uye mhuka dzinogamuchira kuburikidza neinoculation, mhepo uye mvura media [1].Kudzivirirwa kwechirwere chinotapukira kwakakosha sedanho rehutano hweveruzhinji.Nzira huru nhatu dzokurwisa zvirwere zvinotapukira: (1) kudzora kunobva utachiona;(2) kukanganiswa kwenzira yekufambisa;(3) kuchengetedzwa kwehuwandu hwevanhu.Pakati pemazano makuru, kutonga kwenzvimbo yehutachiona kunoonekwa seyakanyanya kukosha zano nekuda kwekureruka kwayo uye mutengo wakaderera.Kukurumidza kuongororwa, kuzviparadzanisa nevamwe, uye kurapwa kwevanhu vane hutachiona kwakakosha, kunoda kukurumidza, kunzwira tsitsi, uye nzira dzekuongorora dzakakwana [2].Ikozvino kuongororwa kwezvirwere zvinotapukira kunowanzo kusanganisa kuongororwa kwekiriniki kunoenderana nezviratidzo uye zviratidzo uye zvidzidzo zvemurabhoritari senge sero tsika uye mamorekuru diagnostics, izvo zvinoda vashandi vakadzidziswa, maitiro akanyanya kushanda, uye inodhura midziyo yekuyedza [3, 4].Kudzivirirwa kwezvirwere zvinotapukira kunoda kukurumidza, kusingadhuri, uye kwakarurama kuongororwa kwenzvimbo, kunyanya munzvimbo dzisina zviwanikwa umo zvirwere zvinotapukira zvakajairika uye zvakakomba [5], pamwe nekurapa murenje kana munhandare yehondo, uko njodzi dzisingatarisirwi..kurapwa kunogumira [6].Muchirevo chechinyorwa chino, microfluidics inyanzvi inobatanidza microelectromechanical systems tekinoroji, nanotechnology, kana sainzi yemidziyo yekunyatso dhiza manipulation [7,8,9,10], ichipa mikana mitsva yekuonekwa-kwe-kutarisira (POCT).) zvirwere zvinotapukira kunze kwezvipatara nemarabhoritari.Kuenzaniswa neyechinyakare-inopedza nguva yekuongorora, microfluidic tekinoroji inopa sampuli uye mari yekuchengetedza yekuongorora mamorekuru panguva yekubuda kwechirwere.Kupararira kwepasi rose kwechirwere chekoronavirus 2019 (COVID-19) kunokonzerwa neyakaoma acute kufema chirwere coronavirus 2 (SARS-CoV-2), saka kukosha kweiyo microfluidics yekudzivirira nenguva uye kutonga kwedenda kunosimbiswa zvakare [11, 12. , 13].Kusiyana nekuongororwa kwechinyakare, microfluidic POCT inoshandisa zvidiki zvinotakurika kubva kubhenji analyzer kusvika kudiki sidestream test mitsetse yekuyedza padhuze nesampling point [14].Iyi bvunzo inoratidzira yakapfava kana isina gadziriro yemuenzaniso, kukurumidza kukwidziridzwa kwechiratidzo, uye inonzwisa masaini kuverenga zvichikonzera kupfupika uye kwakaringana mhinduro mukati memaminitsi.Kuvepo uye kugadzirwa kwakawanda kwezvishandiso zvehutano zvemicrofluidic-based zvakawedzera mari yavo-inoshanda uye yakananga mashandisirwo ekuongorora kunze kwechipatara, pedyo nemurwere, uye kunyangwe kumba.
Pakati pemazano aripo ekuongorora zvirwere zvinotapukira, kuongororwa kwema molecular ndiyo imwe yeanonyanya kunetseka [15, 16].Pamusoro pezvo, kuongororwa kwemamorekuru kunowanzo shandiswa seye goridhe chiyero chekuenderera mberi kwekuonekwa kweCCIDID-19, ichibvumira kutariswa kwakananga kwehutachiona-chaiyo matunhu eRNA kana DNA isati yatanga mhinduro yekudzivirira muviri [17, 18].Muchidzidzo chezvino, tinopa kufambira mberi kwezvino mu microfluidics-based molecular diagnostic process yezvirwere zvinotapukira, kubva pakuona kwezvidzidzo kusvika kune ramangwana remaindasitiri maonero (Fig. 1).Tichatanga nematanho matatu akakosha mukuonekwa kweiyo nucleic acid: pa-chip sample pretreatment, nucleic acid amplification, uye chiratidzo chekuverenga.Isu takazofananidza mhando dzakasiyana dzemicrofluidic mapuratifomu nechimiro chavo uye basa, zvichiratidza hunhu hwakasiyana (simba uye kushaya simba).Digital nucleic acid yekuona inokurukurwa zvakare uye kupihwa semuenzaniso wechizvarwa chechitatu tekinoroji yehuwandu hwehuwandu hwehutachiona hwehutachiona hwemamorekuru.Uye zvakare, akati wandei akajairwa uye achangoburwa ekutengesa POCT zvishandiso zvichaunzwa kuratidza mamiriro azvino eiyo microfluidic POCT musika wekuongororwa kwema molecular.Tichakurukurawo nekutsanangura chiono chedu chezvikumbiro zvemangwana.
Modules ye microfluidic chips ye nucleic acid yekuona inogona kukamurwa kuita zvikamu zvitatu (sampling, recognition, uye signing) zvinoenderana nemabasa avo [19].Pakati pemamodule aya, iyo sampling module inonyanya kuona sampuli lysis uye nucleic acid kubviswa.Iyo sensor module inonyanya kudzora kutendeuka uye kukwidziridzwa kweiyo nucleic acid masaini.Iyo yekusaina module inoona iyo siginecha yakashandurwa uye kugadziriswa neiyo inonzwa module.Zvichienderana nemaitiro ekuona nucleic acids pane chip, isu tichapfupisa akasiyana machipi anogona kuona "yekupinda nekubuda" basa.
Danho rekutanga mukuona nucleic acid kutorwa nucleic acid, kureva kupatsanura yakananga nucleic acid kubva kune yekutanga sampuli.Nucleic acid kubviswa kunoitwa kuchenesa nucleic acids kubva kune mamwe mamorekuru anosvibisa, kuve nechokwadi chekutendeseka kweiyo yekutanga chimiro che nucleic acid mamorekuru, uye kugadzirisa mhedzisiro.Nucleic acid kudhirowa kunoda inodiwa sampuli lysis uye nucleic acid kubatwa, hunhu uye kushanda kwayo zvine simba guru pakutsvagisa nemhedzisiro yekuongorora.Chero ipi neipi yakavanzika mhedzisiro panguva yekubvisa inogona kudzikamisa kuwedzera kuonekwa.Semuyenzaniso, polymerase chain reaction (PCR) uye loop isothermal amplification (LAMP) nzira dzinodzivirirwa nemamwe masara organic solvents akadai se ethanol uye isopropanol mu nucleic acid isolation reagents [20].Liquid-liquid extraction uye solid-phase extraction ndiyo nzira dzinonyanya kufarirwa dzekuparadzanisa nucleic acids [21], zvisinei, mvura-yemvura yakawedzera pane chip yakanyanya kushomeka, sezvo mareagents anoshandiswa mumvura-yemvura kudhirowa anokonzera kukora kweakawanda microfluidic chips. .Pano, isu tinosimbisa microarray-yakavakirwa yakasimba chikamu chekubvisa nzira uye kuenzanisa zvakanakira nekuipira kwavo.
Silicon is substrate material inoenderana nucleic acids nekuda kweiyo biocompatibility, kugadzikana, uye nyore kugadzirisa [22].Zvakakosha, kana yakagadziridzwa nesilika kana zvimwe zvinhu, musanganiswa uyu unoratidza zvivakwa zvekushambadza zvisina kunaka nucleic acids pasi pepH yakaderera, mamiriro emunyu wakanyanya uku uchibuda nepH yakakwirira, yakaderera munyu mhinduro.Kubva pane ichi chiitiko, zvinokwanisika kuchenesa iyo nucleic acid.
Mhando dzakasiyana-siyana dzesilica-based materials dzakashandiswa kunyucleic acid kubudiswa mu microfluidics, zvakadai sesilica beads, powders, microfiber filters, uye silica membranes [23, 24, 25, 26].Zvichienderana nehupfumi hwezvinhu, silicon-based materials inogona kushandiswa mu microcircuits nenzira dzakasiyana.Semuenzaniso, silica granules, powders, uye nanofilters dzekutengesa zvinogona kungoiswa mumapores kana microchannels ye microfluidic chips uye kubatsira kubvisa nucleic acids kubva kusampuli [27, 28, 29].Surface-modified silica membranes inogona zvakare kushandiswa kukurumidza kuchenesa DNA kubva kuhutachiona nemutengo wakaderera.Somuenzaniso, Wang et al.[30] Nekubatanidza denaturing amplification reactions ne vesicle-mediated chain exchange ne silica membranes yakavharwa ne chitosan oligosaccharides, inoshandiswa zvakasiyana-siyana inotakurika sisitimu yakaunzwa iyo yakabudirira kuona 102-108 colony inogadzira zvikamu.(CFU)/ml Vibrio parahaemolyticus., uye kuvapo kweutachiona kwaioneka nyore.Powell et al.[31] Silicon-based microarrays yakabva yashandiswa kuona hepatitis C virus (HCV), human immunodeficiency virus (HIV), Zika virus, uye human papillomavirus uye otomatiki kuparadzira, umo 1.3 μl tortuous microreactor yakagadzirwa kutora RNA mavhairasi.uye kuita in situ amplification.Pamusoro peidzi nzira, pamusoro-yakagadziridzwa silica microcolumns inoitawo basa rakakosha mukubvisa nucleic acid, sezvo geometry uye zvimiro zvechigadziriso zvinowedzera zvakanyanya kudhirowa kushanda.Chen et al.[32] yakakurudzira chikuva chemicrofluidic chekuzviparadzanisa kweRNA yakaderera-yakavakirwa pane amino-yakavharidzirwa silicon microcolumns.Ichi chishandiso chemicrofluidic chinosanganisa mutsara we 0.25 cm2 micropillars pane silicon substrate kuti iwane yakakwira yekubvisa hunyanzvi kuburikidza nepamusoro penzvimbo kusvika kuvhoriyamu reshiyo dhizaini.Chakanakira dhizaini iyi ndechekuti iyo microfluidic mudziyo inogona kusvika kusvika 95% nucleic acid yekubvisa kunyatsoshanda.Aya maitiro esilicon-based anoratidza kukosha kwekukurumidza kuparadzanisa nucleic acids pamutengo wakaderera.Mukubatana ne microfluidic chips, silicon-based extraction strategy haingogone kuwedzera kugona kweiyo nucleic acid yekuona, asi zvakare kufambisa miniaturization uye kubatanidzwa kweanalytical zvishandiso [20].
Magnetic kupatsanura nzira dzinoshandisa magineti zvimedu kuparadzanisa nucleic acids pamberi pekunze kwemagineti.Anowanzo shandiswa magineti particles anosanganisira Fe3O4 kana γ-Fe2O3 magineti particles akaputirwa nesilika, amino uye carboxyl [33,34,35,36].Iyo inosiyanisa magineti zvimedu zvichienzaniswa nesilicon-based SPE nzira ndiko kureruka kwekubata uye kutonga nemagineti ekunze.
Uchishandisa kupindirana kwemagetsi pakati pe nucleic acids uye silica, pasi pemamiriro emunyu wakanyanya uye yakaderera pH, nucleic acids inoshambadzirwa pamusoro pesilica-yakavharidzirwa magineti particles, nepo pasi pemamiriro emunyu wakaderera uye yakakwira pH, mamorekuru anogona kugezwa. zvakare..Silika-yakavharwa nemagineti mabedhi anoita kuti zvibvire kuburitsa DNA kubva muhombe vhoriyamu samples (400 μL) uchishandisa magineti inodzorwa kufamba [37].Sechiratidziro, Rodriguez-Mateos et al.[38] akashandisa tunable magineti kudzora kutamiswa kwemagineti bead kumakamuri akasiyana.Kubva pane silica-yakavharwa magineti particles, 470 makopi/mL eSARS-CoV-2 genomic RNA inogona kutorwa kubva mumarara emvura samples yeLAMP reverse transcription yekuona (RT-LAMP) uye mhinduro inogona kuverengwa mukati meawa imwe.ziso rakashama (Fig. 2a).
Zvishandiso zvinoenderana nemagineti uye porous zvinhu.Conceptual dhizaini yeIFAST RT-LAMP microfluidic mudziyo weSARS-CoV-2 RNA yekuona (yakagadziridzwa kubva [38]).b Centrifugal micro device ye dSPE ye buccal swab nucleic acid (yakashandurwa kubva [39]).c Yakavakirwa-mukati-yega-powered sampuli concentrator uchishandisa FTA® kadhi (yakatorwa kubva [50]).d Fusion 5 sefa bepa rakagadziridzwa nechitosan (yakashandurwa kubva [51]).SARS-CoV-2 yakanyanya acute kufema chirwere coronavirus 2, RT-LAMP reverse transcription loop mediated isothermal amplification, FTA vanotsvaga tekinoroji vanobatana, NA nucleic acid.
Positively charged magnetic particles akanakira kubatanidza phosphate musana we nucleic acid.Pane imwe nzvimbo yemunyu, mapoka asina kunaka phosphate mapoka e nucleic acids anogona kupomerwa zvakanaka pamusoro pemagineti anoumba zvikamu.Naizvozvo, magineti nanoparticles ane hutsinye hwepamusoro uye huwandu hwepamusoro hwemapoka emamino akagadzirwa kuti abvise nucleic acids.Mushure mekuparadzaniswa kwemagineti uye kuvhara, magineti nanoparticles uye DNA complexes inogona kushandiswa zvakananga muPCR, iyo inobvisa kudiwa kwekunatswa uye kunotora nguva kucheneswa uye elution mashandiro [35].Magnetic nanoparticles akaputirwa neasina kunaka carboxyl mapoka akashandiswawo kuparadzanisa nucleic acids adsorbed pamusoro pepamusoro-soro polyethylene glycol uye sodium chloride mhinduro [36].Neaya epamusoro-akagadziridzwa magineti mabedhi, kutorwa kweDNA kunoenderana nekuzotevera kukwidziridzwa.Dignan et al.[39] yakatsanangura otomatiki uye inotakurika centrifugal microfluidic chikuva che nucleic acid pretreatment, ichibvumira vasiri tekinoroji kuti vashandise panzvimbo.Mukuwedzera, kuwirirana kweDNA yakaparadzana neLAMP, nzira yakanyatsokodzera nzvimbo-ye-kutarisira nucleic acid kuongororwa, inowedzera kuratidza zvishoma zvinodiwa zvemichina uye kukodzera kwemavara ekuenzanisa (Fig. 2b).
Magnetic bead nzira dzinopa mukana weiyo otomatiki kudhirowa, mamwe acho aripo mune zvekutengesa otomatiki nucleic acid extractors [KingFisher;ThermoFisher (Waltham, MA, USA), QIAcube® HT;CapitalBio (Beijing, China) uye Biomek®;Beckman (Miami, USA).), Florida, USA)].Zvakanakira kusanganisa magineti mabedhi ane microfluidics anogona kushandiswa kuita otomatiki kudhirowa kwe nucleic acids, izvo zvinogona kufambisira mberi kuvandudzwa kwema molecular diagnostics;zvisinei, musanganiswa wemagineti bead ane microfluidics ichiri kutsamira zvakanyanya pane dzakaoma kudzora masisitimu ekugadzirisa chaiko magineti bead, iyo inotsanangura kufarirwa kwezvigadzirwa zvekutengesa kuve zvakakura uye zvinodhura, izvo zvinomisa kumwe kushandiswa kwemagineti bead muPOCT.
Zvinhu zvakati wandei zvine porous senge yakagadziridzwa nitrocellulose mafirita, Finders Technology Associates (FTA) makadhi, polyethersulfone-based sefa mapepa, uye glycan-yakavharwa zvinhu zvakashandiswawo pakuona nucleic acid [40, 41, 42, 43, 44].Porous fibrous materials senge bepa rine fibrous zvakatanga kushandiswa kupatsanura DNA nekupinza muviri mamorekuru eDNA ane tambo refu ane fiber.Madiki pores anotungamira kune yakasimba yemuviri kurambidzwa kwemamorekuru eDNA, ayo anokanganisa kutorwa kweDNA.Nekuda kwehukuru hwakasiyana hwepore yemapepa ane fibrous, kudhirowa kwakanaka hakugone kusangana nezvinodiwa zveDNA amplification [45, 46].Iyo FTA kadhi ibhizimusi refirita bepa rinoshandiswa mumunda wemushonga wemushonga uye rinoshandiswa zvakanyanya mune dzimwe nzvimbo dzekuongorora mamorekuru.Kuburikidza nekushandiswa kweselulosi sefa bepa rakamisikidzwa nemakemikari akasiyana siyana lyse maseru membranes mumuenzaniso, iyo yakaburitswa DNA inodzivirirwa kubva mukuora kweanosvika makore maviri.Nguva pfupi yadarika, bepa re cellulose rakaiswa rakagadzirwa kuti rionekwe mamorekuru ezvirwere zvakasiyana siyana, kusanganisira SARS-CoV-2, leishmaniasis, uye marariya [47,48,49].HIV mu plasma yakasarudzika yakanyungudutswa zvakananga, uye viral nucleic acid inowedzerwa muFTA® flow membrane yakavakwa mukati me concentrator, iyo inobvumira kugadzirwa kwakanaka kweiyo nucleic acid [50] (Fig. 2c).Dambudziko guru rekuona nucleic acid uchishandisa FTA makadhi nderekuti makemikari akadai seguanidine uye isopropanol anodzivisa kunotevera amplification reactions.Kugadzirisa dambudziko iri, takagadzira Fusion 5 chitosan-modified filter paper, iyo inosanganisa zvakanakira zvese zviri zviviri interlacing eDNA mamorekuru uye fibrous sefa bepa, uye electrostatic adsorption yeDNA pane chitosan-akagadziridzwa makomisheni kuti iwane yakanyatso shanda nucleic acid kutorwa. ..sefa fibers [51] (Fig. 2d).Saizvozvowo, Zhu et al.[52] yakaratidza chitosan-modified PCR nzira inobva pane in situ capillary microfluidic system yekukurumidza kuparadzaniswa uye kuonekwa kweZika virus RNA.Nucleic acids inogona kudsorbed / kunyungudutswa mune yakasanganiswa lysate/PCR svikiro, zvichiteerana, zvichibva pane / off switch pfuma ye chitosan.on and off”, inopindura pH.
Sezvambotaurwa pamusoro apa, mazano aya anosanganisa zvakanakira akasiyana-siyana akasimba chikamu zvinhu uye anowedzera kugona kweiyo nucleic acid kudhirowa mune microfluidics.Mumashandisirwo anoshanda, kushandiswa kwezvinhu izvi muhuwandu hukuru hakuna hupfumi, uye kurapwa kwakakodzera kwepamusoro kana kugadziridzwa kwepamusoro kwezvinhu zvakajairika nezvinhu izvi zvinogonawo kuchengetedza basa razvo.Nokudaro, zvinotendwa kuti kushandiswa kwezvirongwa izvi mushure mekuongorora kwekuongorora kunogona kuderedza mari.
Nucleic acid kuyedza pamapuratifomu emicrofluidic kazhinji inoshandisa mavhoriyamu madiki emuenzaniso (<100 µl), saka inoda kukwidziridzwa kweinonangwa nucleic acids ine maprobes chaiwo ekushandurwa kuita chiratidzo chinokodzera kudzika kudzika (optical, magetsi, uye magineti) [53, 54]. Nucleic acid kuyedza pamapuratifomu emicrofluidic kazhinji inoshandisa mavhoriyamu madiki emuenzaniso (<100 µl), saka inoda kukwidziridzwa kweinonangwa nucleic acids ine maprobes chaiwo ekushandurwa kuita chiratidzo chinokodzera kudzika kudzika (optical, magetsi, uye magineti) [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]. Kana uchiyedza nucleic acids pamapuratifomu emicrofluidic, mavhoriyamu madiki emuenzaniso (<100 µL) anowanzo shandiswa, saka kukwidziridzwa kwechinangwa chenyucleic acids ine maprobe akakosha kunodiwa kuti ishandurwe kuita chiratidzo chakanakira kunotevera kucherechedzwa (Optical, magetsi, uye magineti) [53, 54].微流控 平台 上 的 的 通常 使用 使用 使用 使用 (<100 使用 特定 探针 探针 目标 核酸, 以, 电学 电学 便于 下游 检测 信号 信号 磁学 信号].微流控 平台 上 的 检测 使用 使用 ((<100 特定 ((()., ]. 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]]. Kuonekwa kwe nucleic acids pamapuratifomu emicrofluidic kunowanzo shandisa mavhoriyamu madiki emuenzaniso (<100 μl), izvo zvinoda kukwidziridzwa kwechinangwa chenyucleic acids ine maprobes akakosha kuti ashandure kuita masaini kuti aonekwe (optical, magetsi, uye magineti) [53, 54]] .Nucleic acid amplification mu microfluidics inogonawo kukurumidzira maitiro, kugadzirisa miganhu yekuona, kuderedza sampuli zvinodiwa, uye kunatsiridza kuonekwa kwechokwadi [55, 56].Mumakore achangopfuura, nekuona kwekukurumidza uye kwakaringana kuona, akasiyana nucleic acid amplification nzira dzakashandiswa mune microfluidics, kusanganisira PCR uye kumwe isothermal amplification reactions.Ichi chikamu chichapfupisa nzira dzekuona nucleic acid kubva pane microfluidic masisitimu.
PCR kuenzanisa kweDNA kudzokorodza maitiro echinhu chipenyu, dzidziso yacho inotsanangurwa zvakadzama kumwe kunhu uye haizokurukurwa pano.PCR inogona kukwidziridza huwandu hushoma hwechinangwa cheDNA/RNA pamwero weexponential, zvichiita kuti PCR ive chishandiso chine simba chekuona nekukurumidza nucleic acids.Mumakumi emakore achangopfuura, akawanda anotakurika microfluidic madivayiri akashongedzerwa nePCR ekupisa mabhasikoro masisitimu akagadzirwa kuti asangane nezvinodiwa zvekunongedza-kwe-kutarisira diagnostics [57, 58].On-chip PCR inogona kukamurwa kuita mhando ina (yakajairika, inoenderera kuyerera, spatially switched, uye convective PCR) zvinoenderana neyakasiyana tembiricha nzira dzekudzora [59].Somuenzaniso, Gee et al.[60] vakagadzira yakananga reverse transcription quantitative PCR (RT-qPCR) nzira yavo pachavo microfluidic papuratifomu ye multiplex kuonekwa kweSARS-CoV-2, furuwenza A uye B mavhairasi pahuro swab samples (Fig. 3a) .Park et al.[61] yakavaka chip yakareruka pathogen yekuongorora nekubatanidza firimu rakaonda PCR, electrodes, uye chigunwe-chinoshandiswa polydimethylsiloxane-based microfluidic module.Nekudaro, ese ari maviri mabasa anosanganisira izvo zvakajairika zvikanganiso zveyakajairika PCR.PCR inoda kuchovha bhasikoro, iyo inodzikamisa imwe mudziyo miniaturization uye kuderedzwa nguva yekuyedza.
Iko kuvandudzwa kwekuenderera mberi kuyerera kwakavakirwa microfluidic uye nzvimbo-yakachinja PCR yakakosha kugadzirisa nyaya iyi.Uchishandisa chiteshi chenyoka refu kana kuti pfupi yakatwasuka chiteshi, inoenderera ichiyerera PCR inogona kupa kukurumidza kukwidziridzwa nekushingairira kutenderera reagents munzvimbo nhatu dzekutanga kupisa ine off-chip pombi.Uku kuvhiya kunobudirira kudzivirira chikamu chekuchinja pakati pekuchinja kwakasiyana kwekushisa uye nokudaro kunoderedza zvakanyanya nguva yekuedza [62] (Fig. 3b).Mune imwe ongororo yakaitwa naJung et al.[63] yakaronga itsva rotary PCR genetic analyzer iyo inobatanidza maitiro ekugadzirisa uye kuyerera PCR ye ultrafast uye multiplex reverse transcription PCR (Fig. 3c).Nokuda kweiyo nucleic acid amplification, PCR microchip ichatenderedzwa kuburikidza nematombo matatu ekupisa pakupisa kwakasiyana: 1. Denaturation block 94 ° C, 2. Annealing block pa 58 ° C, 3. Chivharo chekuwedzera pa 72 ° C.
Kushandiswa kwePCR mune microfluidics.Schematic inomiririra ye dirRT-qPCR pane microfluidic papuratifomu (yakagadziriswa kubva [60]).b Schematic inomiririra yeinoenderera ichiyerera PCR microarray yakavakirwa pane nyoka chiteshi (yakagadziriswa kubva [62]).c Schematic inomiririra ye rotary PCR genetic analyzer, inosanganisira microchip, matatu ekudziya mabhuroki uye stepper mota (yakashandurwa kubva [63]).d Dhiagiramu ye thermoconvection PCR ine centrifugation uye setup (yakagadziriswa kubva [64]).DirRT-qPCR, yakananga kuwanda reverse transcription polymerase chain reaction
Uchishandisa macapillaries uye zvishwe kana kunyange mahwendefa matete, convection PCR inogona kukurumidza kukwidziridza nucleic acids neakasikwa emahara thermal convection pasina kudiwa kwepombi yekunze.Semuyenzaniso, cyclic olefin polymer microfluidic platform yakagadzirwa payakagadzirwa inotenderera inopisa nhanho inoshandisa kupisa bhasikoro ne centrifugation muPCR loop microchannel [64] (Fig. 3d).Mhinduro yemhinduro inofambiswa neiyo thermal convection, iyo inoramba ichitsinhana yakakwira uye yakaderera tembiricha mune microchannel ine annular chimiro.Iyo yese amplification process inogona kupedzwa mumaminitsi gumi ine muganho wekuona we70.5 pg/channel.
Sezvinotarisirwa, kukurumidza PCR chishandiso chine simba chakazara chakabatanidzwa-sample-response molecular diagnostic and multiplex analysis systems.Rapid PCR inoderedza zvakanyanya nguva inodiwa kuona SARS-CoV-2, iyo inobatsira kudzora kunoshanda kweCOVID-19 denda.
PCR inoda yakaoma thermal cycler iyo isina kukodzera POCT.Munguva pfupi yapfuura, isothermal amplification maitiro akashandiswa kune microfluidics, kusanganisira asi isina kuganhurirwa kuLAMP, recombinase polymerase amplification (RPA), uye amplification yakavakirwa nucleic acid sequences [65,66,67,68].Nematanho aya, nucleic acids inokwidziridzwa patembiricha isingaperi, ichigonesa kugadzirwa kwemutengo wakaderera, wakanyanya kutaridzika anotakurika POCT zvishandiso zvemamolecular diagnostics.
High-throughput microfluidics-based LAMP assays inobvumira kuonekwa kwakawanda kwezvirwere zvinotapukira [42, 69, 70, 71].Mukubatana ne centrifugal microfluidic system, LAMP inogona kuenderera mberi nekufambisa otomatiki ye nucleic acid yekuona [69, 72, 73, 74, 75].I-spin-and-react SlipChip yakagadzirwa nokuda kwekuona kuonekwa kwemabhakitiriya akawanda akafanana achishandisa LAMP [76] (Fig. 4a).Paunenge uchishandisa yakagadziridzwa LAMP muyedzo, iyo fluorescence siginecha-kune-ruzha chiyero yaive ingangoita 5-kupeta, uye muganho wekuona wakasvika 7.2 makopi / μl egenomic DNA. Uyezve, kuvepo kwezvishanu zvinowanzoitika hutachiona hwehutachiona hwekudya, hunosanganisira Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis uye Vibrio parahaemolyticus, zvakaonekwa zvichibva pane iyo nzira mu <60 min. Uyezve, kuvepo kwezvishanu zvinowanzoitika hutachiona hwehutachiona hwekudya, hunosanganisira Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis uye Vibrio parahaemolyticus, zvakaonekwa zvichibva pane iyo nzira mu <60 min.Uyezve, kuvapo kwezvirwere zvishanu zvakajairika zvebhakitiriya zvegumbo rekudya, kusanganisira Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis uye Vibrio parahaemolyticus, yakaonekwa pachishandiswa nzira iyi isingasviki maminetsi makumi matanhatu.此外, 基于 该 方法 方法 <60 分钟 内 可 视化 五 五 种 常见 常见 原体 原体 原体 原体 原体 原体 原体此外, 基于 该 方法 方法 <60 分钟 内 视化 了 五 种 常见 常见 的 的 存在, 包括, 包括,弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 HIPPamusoro pezvo, kuvepo kwezvirwere zvishanu zvakajairika zvemabhakitiriya emudumbu, kusanganisira Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvius, uye Vibrio parahaemolyticus, yakaonekwa pachishandiswa nzira iyi isingasviki maminetsi makumi matanhatu.
Zvakanakira LAMP mune microfluidics zvinosanganisira, pakati pezvimwe, kukurumidza kupindura uye miniaturized kuona.Nekudaro, nekuda kwekuita tembiricha (yakatenderedza 70 ° C), maaerosols anogadzirwa panguva yeLAMP, zvichikonzera chiyero chepamusoro chenhema.Assay chaiyo, dhizaini yekutanga, uye tembiricha yekudzora inodawo kugadziridzwa yeLAMP.Pamusoro pezvo, madhizaini echipi anoisa akawanda anotariswa kuona pane imwechete chip akakosha zvikuru uye anofanirwa kuvandudzwa.Pamusoro pezvo, LAMP inokodzera kucherechedzwa kwezvinangwa zvakawanda zvakabatanidzwa mune imwe chip, iyo yakakosha zvikuru, asi kuchine nzvimbo yakawanda yebudiriro.
Iyo yakakwirira yenhema yakanaka mwero weLAMP inogona kuderedzwa pamwe neRPA, sezvo yakaderera kuita tembiricha (~ 37 ° C) inoguma mune mashoma mashoma evaporation matambudziko [77].Muchirongwa cheRPA, maviri akatarisana ekutanga anotanga DNA synthesis nekusunga kune recombinase uye kukwidziridzwa kunogona kupedzwa mukati memaminitsi gumi [78,79,80,81].Naizvozvo, iyo yese RPA maitiro inokurumidza kupfuura PCR kana LAMP.Mumakore achangopfuura, tekinoroji yemicrofluidic yakaratidzwa kuwedzera kukurumidza uye kurongeka kweRPA [82,83,84].Somuenzaniso, Liu et al.[85] yakagadzira microfluidic yakabatanidzwa lateral flow polymerase recombinase amplification assay yekukurumidza uye inonzwisisika yeSARS-CoV-2 nekubatanidza reverse transcription RPA (RT-RPA) uye universal lateral flow test strip yekuona system.mune imwechete microfluidic system.Mufananidzo 4b).Muganho wekuonekwa i1 kopi/µl kana makumi matatu kopi/sample, uye kuona kunogona kupera mukati memaminitsi makumi matatu.Kong et al.vakagadzira mudziyo unopfekwa wemicrofluidic.[86] akashandisa tembiricha yemuviri uye nharembozha-yakavakirwa fluorescence yekuongorora system kuti ikurumidze kuona zvakananga HIV-1 DNA uchishandisa RPA (Mufananidzo 4c).Iyo inopfekwa yeRPA assay inoona zana remakopi / mL yechinangwa chakatevedzana mukati memaminitsi makumi maviri nemana, zvichiratidza kugona kukuru kwekukurumidza kuongororwa kwevacheche vane HIV-1 mune zvigadziriso-zvishoma.
Isothermal amplification mu-point-of-care test (POCT).Kuvandudza uye kugadzirwa kwe spin uye kuita SlipChip.Mushure mekuputika kweplasma, machipisi epamusoro uye epasi akaunganidzwa neseti yenzungu kuti agadzire chip yekupedzisira (yakagadziriswa kubva [76]).b Schematic yeMI-IF-RPA system yeCOVID-19 yekuona (yakagadziridzwa kubva [85]).c Schematic yekupfekeka RPA bvunzo yekukurumidza kuona yeHIV-1 DNA (yakagadziriswa kubva [86]).SE Salmonella enterica, VF Vibrio fluvius, VP Vibrio parahaemolyticus, BC Bacillus cereus, EC Escherichia coli, FAM carboxyfluorescein, human immunodeficiency virus HIV, RPA recombinase polymerase amplification, LED mwenje emitting diode, MI-Flumeraro reMicrofluoreg-IF-IF-FImeral Diode Mukati meMicrofluoreg-IF-IF-IF-FImerase. Amplification
Microfluidic-based RPA iri kukura nekukurumidza, zvisinei, mutengo wekugadzira chip uye mashandisiro ekuita wakanyanya uye unofanirwa kuderedzwa kuti uwedzere kuwanikwa kweiyi tekinoroji.Uye zvakare, iyo yakanyanya senitivity yeRPA inogona kukanganisa kukwidziridzwa kwezvinhu zvisiri-chaiyo zvigadzirwa, kunyanya pamberi pekusvibiswa.Izvi zvinogumira zvinogona kukanganisa mashandisirwo eRPA mune microfluidic masisitimu uye akakodzera kuwedzera optimization.Yakagadzirwa zvakanaka maprimers uye probes ezvinangwa zvakasiyana anodiwa zvakare kuvandudza kugona kweRPA-yakavakirwa microfluidic mazano muPOCT.
Cas13 neCas12a vane hunyanzvi hwekutsemura nucleic acids uye nekudaro inogona kugadzirwa sematurusi ekuona uye ekuongorora.Cas13 neCas12a zvinobatiswa pakusunga kunanga DNA kana RNA, zvichiteerana.Kana yangoitwa, puroteni inotanga kutsemura mamwe ari padyo nucleic acids, mushure meiyo inotungamira maRNA anonangana pathogen-chaiwo nucleic acids anogona kubaya akadzimwa efluorescent probes uye kuburitsa fluorescence.Kubva pane iyi dzidziso, Kellner et al.[87] yakagadzira Cas13-based method [Specific High-sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK)], uye Broughton et al.[88] yakagadzira imwe nzira yakavakirwa paCas12a [CRISPR Trans Reporter yakanangana neDNA endonuclease (DTECR)].
Mumakore achangopfuura, nzira dzakasiyana dzekuona nucleic acids dzakavakirwa paCRISPR dzakaonekwa [89, 90].Yakajairika CRISPR yakavakirwa nzira dzinowanzo kutora nguva uye kushanda nesimba nekuda kwemaitiro akawanda anosanganisira nucleic acid kudhirowa, amplification uye CRISPR kuona.Kuratidzwa kwezvinwiwa kumhepo kunogona kuwedzera mukana wekunyepa kwemhedzisiro.Tichifunga zviri pamusoro, CRISPR-yakavakirwa masisitimu ari kuda zvakanyanya optimization.
Iyo pneumatically inodzorwa microfluidic chikuva inogona kuita 24 ongororo yakafanana yakagadziridzwa yeCRISPR-Cas12a uye CRISPR-Cas13a yekuongorora maapplication [91].Iyo sisitimu ine fluorescence yekuona mudziyo iyo inonzvenga nucleic acid amplification uye inongoerekana yaona femtomolar DNA neRNA samples.Chen et al.[92] yakasanganiswa recombinase amplification neCRISPR-Cas12a system mu centrifugal microfluidics (Fig. 5a).Iri basa rinokunda kuoma kwekubatanidza maitiro maviri aya nekuti Cas12a inogona kugaya mutumwa DNA uye inhibisa maitiro ekusimudzira.Mukuwedzera, Chen et al.[92] nekuwedzera pre-yakachengetedza maitiro reagents mune centrifugal microfluidic control kuti ipedze otomatiki maitiro ese.Mune rimwe basa, Silva et al.[93] yakagadzira nzira yekuongorora pasina CRISPR / Cas12a amplification uye smartphone kuti ione SARS-CoV-2 (Fig. 5b).Iyi assay, inozivikanwa senhare-yakavakirwa amplification-yemahara sisitimu, inosanganisira CRISPR/Cas-inotsamira enzyme yakavakirwa pa smartphone kuona kwecatalase-yakagadzirwa bubble masaini mumakrofluidic chiteshi.Sensitive yekuona isingasviki makumi mashanu makopi/µl ye nucleic acid isina pre-amplification, iyo yese maitiro kubva kumuenzaniso jekiseni kuenda kuchiratidzo kuverenga inotora chete 71 maminetsi.
Nucleic acid yekuona nzira dzakavakirwa paCRSPR.Centrifugal POCT yekubatanidzwa kwema molecular diagnostics yakavakirwa paCRISPR (yakashandurwa kubva [92]).b Kukudziridzwa kweCASCADE bvunzo ye-smartphone-based ongororo yeSARS-CoV-2 (yakagadziriswa kubva [93]).RAA recombinase amplification, PAM iri pedyo neprotospacer motif, CRISPR yakabatanidzwa pfupi palindromic inodzokorora nguva nenguva, CASCADE system isina kukwidziridzwa kwefoni neCRISPR / CAS-inotsamira enzymes, 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride EDC
Seyekupedzisira nhanho yekuona nucleic acid, kuona chiratidzo kunoratidza zvakananga mhedzisiro yekuongorora uye chinhu chakakosha mukugadzirwa kwePOCT inoshanda, inonzwisisika, uye yakarurama.Zviratidzo zvinogona kuverengwa uchishandisa nzira dzakasiyana senge fluorescent, electrochemical, colorimetric uye magineti mazano.Muchikamu chino, tinotsanangura chikonzero chemaitiro ega ega uye kuenzanisa kuongororwa kwema molecular ezvirwere zvinotapukira mu microfluidics.
Fluorescence-based mazano anoshandiswa zvakanyanya kuPOCT kuongororwa kwezvirwere zvinotapukira nekuda kwezvakanaka zvakanakisa zvekunzwa kwakanyanya, mutengo wakaderera, kusununguka kwekushanda, uye ongororo yekutarisisa [94, 95].Aya marongero anoshandisa akanyorwa fluorophores senge fluorescent dhayi uye nanomaterials kugadzira chiratidzo chinooneka (fluorescence kuwedzera kana kudzima).Kuwana uku kunoratidza kuti fluorescence-based strategy inogona kukamurwa kuita yakananga fluorescent labeling, chiratidzo-pa, uye chiratidzo-kudzima fluorescent yekuona [96].Direct fluorescent label yekuona inoshandisa akakosha mavara efluorescent kuti aise chaiwo ligands anoburitsa huwandu hwakati hwefluorescence kana zvakasarudzwa zvakasungwa kune chinangwa.Nokuda kwechiratidzo-based fluorescence kuonekwa, kunaka kwechiratidzo chefluorescent kune hukama hwakanaka nehukuru hwekufarira.Fluorescence intensity haigoneki mukushayikwa kwechinangwa uye inoonekwa kana huwandu hwakakwana hwechinangwa huripo.Sezvineiwo, kusimba kwefluorescence kunoonekwa ne "signal-off" fluorescence inversely proportional to the amount of target, kutanga kusvika pakukosha kwepamusoro uye zvishoma nezvishoma kuderera sezvo chinangwa chinowedzera.Semuyenzaniso, kushandisa CRISPR-Cas13a chinangwa-inotsamira trans-cleavage michina, Tian et al.[97] yakagadzira hurongwa hwekucherechedzwa hutsva hwekuona maRNA anonzvenga nyora reverse zvakananga (Fig. 6a).Pakusunga kune inowirirana chinangwa RNAs, iyo CRISPR-Cas13-RNA yakaoma inogona kuvhurwa, ichikonzera transcollateral cleavage neasina-chaiyo mutori wenhau RNAs.Iyo fluorescent yakanyorwa reporter [fluorophore (F)] inodzimwa ne quencher (Q) yakasimba uye fluoresces kana yakatsemurwa neyakagadziriswa yakaoma.
Iyo mukana wekuona electrochemical kukurumidza kuona, kugadzirwa kuri nyore, mutengo wakaderera, nyore kutakura uye otomatiki kudzora.Iyo inzira ine simba yekuongorora yePOCT application.Kubva pane graphene munda-effect transistors Gao et al.[98] yakagadzira nanobiosensor ye multiplex yekuona maantigen echirwere cheLyme kubva kuBorrelia burgdorferi mabhakitiriya ane muganhu wekuona we2 pg / mL (Fig. 6b).
Colorimetric assays yakashandiswa muPOCT application, ichibatsirwa kubva kune zvakanakira kutakura, mutengo wakaderera, nyore kugadzirira, uye kuverenga kwekuona.Kuonekwa kwemavara kunogona kushandisa oxidation ye peroxidase kana peroxidase-like nanomaterials, kuunganidzwa kwemananomaterials, uye kuwedzerwa kwedhayi dzinoratidza kushandura ruzivo nezvekuvapo kwechinangwa nucleic acids kuita shanduko yemavara inooneka [99, 100, 101].Zvinonyanya kukosha, nanoparticles yegoridhe inoshandiswa zvakanyanya mukuvandudza nzira dzemavara, uye nekuda kwekugona kwavo kukurumidza uye kwakakosha shanduko yeruvara, pane kuwedzera kufarira mukuvandudzwa kwePOCT colorimetric mapuratifomu e in situ kuongororwa kwezvirwere zvinotapukira [102].Nechigadzirwa che centrifugal microfluidic device [103], zvirwere zvinokonzerwa nezvokudya mumasampuli emukaka wakasvibiswa zvinogona kuonekwa pakarepo pamwero wemasero ebhakitiriya e10, uye zvigumisiro zvinogona kuverengwa nekuona mukati memaminitsi e65 (Fig. 6c).
Magnetic sensing matekiniki anogona kunyatsoona vanalyte vachishandisa magineti zvinhu, uye pakave nekufarira kwakanyanya muPOCT application mumakumi emakore achangopfuura.Magnetic sensing matekiniki ane mamwe mabhenefiti akasarudzika senge mutengo wakaderera wemagineti pane kudhura zvinhu zvemaziso.Zvisinei, kushandiswa kwemagineti kunovandudza kuonekwa uye kunoderedza nguva yekugadzirira sampuli [104].Mukuwedzera, migumisiro yemagnetic probing inoratidza hukuru hwepamusoro, manzwiro, uye yakakwirira-signal-to-noise ratio nekuda kwekusakosha kwemagineti kumashure kwezviratidzo zvehupenyu [105].Sharma et al.yakabatanidza magnetic tunnel junction yakavakirwa biosensor mune inotakurika microchip chikuva.[106] yekuongorora kwakawanda kwezvirwere (Fig. 6d).Biosensors inocherekedza subnanomolar nucleic acids yakaparadzaniswa nehutachiona.
Yakajairika nzira yekuona masaini.Pfungwa yekuona hyperlocalized yeCas13a (yakashandurwa kubva [97]).b Graphene nanobiosensor FET pamwe chete neLyme GroES scFv (yakagadziridzwa kubva [98]).c Colorimetric zviratidzo zvekuonekwa kwe multiplex yezvipembenene zvezvokudya mu centrifugal microfluidic chip: No. 1 uye No. .d Biosensor yakavakirwa pane magineti tunnel junction, kusanganisira chikuva, yakavakirwa-mukati inovharira amplifier, control unit, uye magetsi ekugadzira chiratidzo / kutora (yakagadziriswa kubva [106]).GFET Graphene FET, Escherichia coli, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus, Vibrio parahaemolyticus, Listeria monocytogenes, PC PC, PDMS Dimethicone, PMMA polymethyl methacrylate
Pasinei neunhu hwakanakisa hwenzira dzekuona dziri pamusoro, dzichiri nekuipa.Idzi nzira dzinofananidzwa (tafura 1), kusanganisira mamwe maapplication ane ruzivo (zvakanakira nezvayakaipira).
Nekuvandudzwa kwe microfluidics, microelectromechanical systems, nanotechnology uye zvinhu zvesayenzi, kushandiswa kwe microfluidic chips pakuona zvirwere zvinotapukira kunoramba kuchifambira mberi [55,96,107,108].Kunyatso shandiswa kwemidziyo midiki uye zvinwiwa zvinobatsira mukurongeka kwekuongorora uye kuita mari.Naizvozvo, kune imwe budiriro, kuedza kwakaitwa kukwirisa nekusimudzira machipisi, zvichikonzera akasiyana microfluidic machipi ane akasiyana zvimiro uye mabasa.Pano isu tinosuma muchidimbu akati wandei akajairwa mhando dzemicrofluidic mapuratifomu uye tichienzanisa maitiro avo (zvakanakira nezvakashata).Pamusoro pezvo, mizhinji yemienzaniso yakanyorwa pazasi inotarisa zvakanyanya pakurwisa SARS-CoV-2.
LOCCs ndiyo yakajairika miniaturized yakaoma analytical masisitimu uye mashandiro avo akanyanya miniaturized, akabatanidzwa, otomatiki uye akafananidzwa kubva kumuenzaniso jekiseni uye kugadzirira, kuyerera kwekudzora uye kuona mvura [109, 110].Zvinwiwa zvinoshandiswa kuburikidza nekunyatsogadzirwa geometry uye kupindirana kwemaitiro mazhinji emuviri senge pressure gradients, capillary action, electrodynamics, magnetic fields uye acoustic waves [111].LOCC inoratidza mabhenefiti akanakisa mukutarisisa kwepamusoro uye kutariswa kwakawanda, nekukurumidza kuongororwa kumhanya, saizi diki yemuenzaniso, kuderera kwesimba rekushandisa, uye kukwirira kwepamusoro uye kushanda zvakanaka;zvisinei, midziyo yeLOCC yakanyanya kutetepa, uye kugadzira, kurongedza, uye kupindirana.Nekudaro, kuwanda uye kushandisazve kutarisana nematambudziko makuru [96].Kuenzaniswa nemamwe mapuratifomu, LOCC ine mabhenefiti akasiyana maererano nehukuru hwemashandisirwo akasiyana uye yakanyanya kunaka tekinoroji kuenderana, asi zvayakaipira zviri pachena zvakare, zvinoti kuomarara kwepamusoro uye kusadzokororwa zvakanaka.Kutsamira pamapombi ekunze, ayo anowanzo kuwanda uye anodhura, anowedzera kumisa kushandiswa kwawo muPOCT.
Munguva yeCOVID-19 kubuda, LOCC yakagamuchira kutariswa kwakawanda.Panguva imwecheteyo, kune akati wandei machipisi anosanganisa akati wandei matekinoroji.Semuenzaniso, ma smartphones ave kushandiswa zvakanyanya seanotakurika analytics zvishandiso uye ane mukana mukuru wekubatanidzwa kweLOCC.Zuva et al.[21] akagadzira microfluidic chip iyo inobvumira kuwanda kwakananga nucleic acid kutevedzana kwezvirwere zvishanu, kusanganisira SARS-CoV-2, vachishandisa LAMP uye vakazviongorora vachishandisa smartphone mukati meawa 1 mushure mekupera kwekuita.Semumwe muenzaniso, Sundah et al.[112] yakagadzira molecular switch [catalytic amplification by molecular transition state switch (CATCH)] yekuona yakananga uye nekunzwa kweSARS-CoV-2 RNA zvinangwa uchishandisa mafoni. CATCH inoenderana neinotakurika LOCC uye inowana hukuru hwekuita (inenge 8 RNA makopi/μl; <1 h pakamuri tembiricha) [112]. CATCH inoenderana neinotakurika LOCC uye inowana hukuru hwekuita (inenge 8 RNA makopi/μl; <1 h pakamuri tembiricha) [112]. CATCH совместим с портативным LOCC и обеспечивает превосходную производительность (примерно 8 копий РНК/мкл; < 1 ч при компение 12]. CATCH inoenderana neinotakurika LOCC uye inopa yakanakisa throughput (inenge 8 RNA makopi/µl; <1 h pakamuri tembiricha) [112]. CATCH 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 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 портативными LOCC и обладает превосходной производительностью (примерно 8 копий РНК/мкл; < 1 часа при комнат1т12 тем].) CATCH inowirirana neinotakurika LOCCs uye ine mashandiro akanaka (anenge 8 RNA makopi/µl; <awa 1 patembiricha yekamuri) [112].Pamusoro pezvo, midziyo yeLOCC yekuongorora mamorekuru inoshandisawo mamwe masimba ekutyaira senge vacuum, kutambanudza, uye minda yemagetsi.Kang et al.[113] yakaratidza chaiyo-nguva, yekupedzisira-inokurumidza nanoplasma-on-a-chip PCR yekukurumidza uye kuwanda kuongororwa kweCCIDID-19 mumunda uchishandisa vacuum plasmonic liquid PCR chip.Li et al.[114] yakazogadzira yakatambanudzwa-inotyairwa microfluidic chip iyo yakagonesa kuongororwa kweCOVID-19.Iyi puratifomu inoshandisa iyo RT-LAMP amplification system kuona kana sampuli yakanaka kana yakaipa.Zvadaro, Ramachandran et al.[115] akawana zvakakodzera magetsi emunda gradients achishandisa isotachophoresis (ITP), yakasarudzwa ion inotarisa nzira inoshandiswa mu microfluidics.NeITP, chinangwa cheRNA kubva mumbishi nasopharyngeal swab samples inogona kucheneswa otomatiki.Zvadaro Ramachandran et al.[115] Kubatanidza iyi kucheneswa kweITP neITP-enhanced LAMP uye CRISPR assays akaonekwa SARS-CoV-2 mumunhu nasopharyngeal swab uye kiriniki mienzaniso mune angangoita 35 maminetsi.Mukuwedzera, pfungwa itsva dzinogara dzichibuda.Jadhav et al.[116] yakaronga chirongwa chekuongorora chakavakirwa pamusoro-chakakwidziridzwa cheRaman spectroscopy pamwe chete neine microfluidic mudziyo une vertical oriented goridhe / sirivha-yakavharwa kabhoni nanotubes kana inoraswa electrospun micro/nanotubes.Membrane-inoshanda yakavakirwa-mukati mafirita microchannels anoraswa.Chishandiso chinoshambadza mavhairasi kubva kumvura dzakasiyana-siyana dzemuviri / kubuda senge mate, nasopharynx uye misodzi.Nekudaro, iyo hutachiona titer yakawanda uye hutachiona hunogona kunyatsozivikanwa neiyo Raman siginecha.
LOAD is centrifugal microfluidic platform umo maitiro ese anodzorwa ne frequency protocol inotenderera microstructured substrate [110].Iyo LOAD mudziyo unoratidzwa nekushandisa centrifugal simba sechinhu chakakosha chekutyaira.Liquids zvakare iri pasi pe capillary, Euler uye Coriolis mauto.Uchishandisa centrifuge mudziyo, ongororo inoitwa mukuenderera kwemvura kushanda kubva kune radial mukati kuenda kunze chinzvimbo, kubvisa kudiwa kwekuwedzera ekunze tubing, pombi, actuators, uye anoshanda mavharuvhu.Muchidimbu, nzira imwe chete yekudzora inorerutsa kushanda.Masimba anoshanda pamvura mune imwecheteyo microfluidic chiteshi pachinhambwe chimwe kubva panzvimbo yekuremedza akaenzana, izvo zvinoita kuti zvikwanise kudzokorora chimiro chechiteshi.Saka, midziyo yeLOAD iri nyore uye inodhura kugadzira nekugadzira pane yakajairwa LOCC midziyo, nepo mabatiro acho akazvimiririra uye akafananidzwa;zvisinei, nekuda kwesimba rakakwirira remagetsi emidziyo ye centrifugal, iripo chip zvinhu ishoma uye mavhoriyamu madiki akaoma.kumota.Panguva imwecheteyo, michina yakawanda yeLOAD yakagadzirirwa kushandiswa kumwe chete, iyo inodhura pakuona kukuru [96, 117, 118, 119].
Mumakumi emakore achangopfuura, LOAD, inoonekwa seimwe yeanonyanya kuvimbisa microfluidic zvishandiso, yakagamuchira tarisiro huru kubva kune vaongorori nevagadziri.Nekudaro, LOAD yakawana kugamuchirwa kwakawanda uye yakashandiswa kuongororwa kwema molecular ehutachiona hwehutachiona [120, 121, 122, 123, 124], kunyanya panguva yekubuda kweCOVID-19.Semuenzaniso, pakupera kwa2020, Ji et al.[60] yakaratidza yakananga RT-qPCR muyedzo wekukurumidza uye otomatiki kuona kwakafanana kweSARS-CoV-2 uye furuwenza A uye B hutachiona muhuro swab mienzaniso.Ipapo Xiong et al.[74] yakaratidza LAMP-yakabatanidzwa discoid microfluidic chikuva chekukurumidza, kwakaringana, uye panguva imwe chete kuonekwa kwevanhu vanomwe vanofema coronaviruses, kusanganisira SARS-CoV-2, mukati memaminitsi makumi mana.Mukutanga kwa2021, de Oliveira et al.[73] yakaratidza polystyrene toner centrifugal microfluidic chip, inoshanda nemaoko nemunwe rotator, yeRT-LAMP molecular diagnostic yeCOVID-19.Zvadaro, Dignan et al.[39] yakaratidza otomatiki inotakurika centrifuge microdevice yekucheneswa kweSARS-CoV-2 RNA zvakananga kubva kune buccal swab zvikamu.Medved et al.[53] yakakurudzira inline SARS-CoV-2 aerosol sampling system ine vhoriyamu diki inotenderera microfluidic fluorescent chip ine muganho wekuona wegumi makopi/μL uye diki kutenderera chikumbaridzo chemaminitsi gumi nemashanu.Suarez et al.[75] nguva pfupi yadarika yakashuma kuvandudzwa kweiyo yakabatanidzwa modular centrifugal microfluidic chikuva chekuonekwa kwakananga kweSARS-CoV-2 RNA mukupisa-inactivated nasopharyngeal swab samples uchishandisa LAMP.Iyi mienzaniso inoratidza mabhenefiti makuru uye chivimbiso cheLOAD mune mamolecular diagnostics eCOVID-19.
Muna 1945 Muller naClegg [125] vakatanga kuratidza nzira dzemicrofluidic pabepa vachishandisa sefa bepa neparafini.Muna 2007, boka reWhitesides [126] rakagadzira yekutanga inoshanda mapepa chikuva cheprotein uye glucose yekuongorora.Pepa rave rakanakira substrate ye microfluidics.Iro bepa rine zvinhu zvemukati senge hydrophilicity uye porous chimiro, yakanakisa biocompatibility, huremu huremu, kuchinjika, kupeta, mutengo wakaderera, nyore kushandisa uye nyore.Classical µPADs ine hydrophilic/hydrophobic zvimiro zvakavakwa pamapepa substrates.Zvichienderana nehutatu-dimensional chimiro, μPADs inogona kukamurwa kuita maviri-dimensional (2D) uye matatu-dimensional (3D) μPADs.2D µPADs inogadzirwa nekugadzira hydrophobic miganhu kuti iite microfluidic chiteshi, nepo 3D µPADs inowanzo gadzirwa kubva kune akaturikidzana e2D microfluidic bepa, dzimwe nguva nekupeta bepa, nzira dzekutsvedza, nzira dzakavhurika, uye 3D kudhinda [96].Mvura ine mvura kana yebhayoloji pa μPAD inonyanya kudzorwa necapillary simba pasina kunze kwesimba sosi, kufambisa kusati kwachengetedzwa kwemareagents, sampuli kubata, uye multiplex kuonekwa.Nekudaro, kutonga kwakaringana kwekuyerera uye kuwonekwa kwakawanda kunokanganiswa nekusakwana kwekuona kumhanya, kunzwa, uye reusability [96, 127, 128, 129, 130].
Senge isina kujairika microfluidic chikuva, μPAD yakasimudzirwa zvakanyanya uye yakagadziridzwa kuti iongororwe mamorekuru ezvirwere zvinotapukira zvakaita seHCV, HIV, uye SARS-CoV-2 [131, 132].Nekusarudza uye kunzwisiswa kweHCV, Tengam et al.[133] yakagadzira novel biosensor yakavakirwa pabepa refluorescent uchishandisa yakanyanya kujeka nucleic acid probe yakavakirwa papyrrolidinyl peptide.Nucleic acids inovharika zvisingaite pachidimbu oxidized cellulose bepa nekudzoreredza alkylation pakati pemapoka emaamino nemapoka ealdehyde, uye kuonekwa kunobva pafluorescence.Aya masaini anogona kuverengerwa neyakagadzirwa gadget ine inotakurika fluorescent kamera yakabatana nenharembozha kamera.Zvadaro, Lu et al.[134] akagadzira bepa-based flexible electrode yakavakirwa nickel/goridhe nanoparticles/carbon nanotubes/polyvinyl alcohol organometallic framework macomposites ekutarirwa kweHIV neDNA hybridization uchishandisa methylene yebhuruu seDNA redox chiratidzo.Munguva pfupi yapfuura, Chowdury et al.[135] yakaratidza fungidziro yepuratifomu dhizaini yenzvimbo-ye-kutarisira µPAD yekuyedza uchishandisa mate emurwere akasvibira musanganiswa neLAMP uye inotakurika yekufungidzira tekinoroji yeCOVID-19 analyte yekuona.
Lateral kuyerera bvunzo inotungamira zvinwiwa necapillary masimba uye kudzora mafambiro emvura nekunyorova uye hunhu hweporous kana microstructured substrates.Iyo lateral inoyerera michina ine sampuli, conjugate, incubator uye yekuona, uye inonwisa mapadhi.Iyo nucleic asidhi mamorekuru muLFA anoziva mabhaidhi chaiwo akafanochengetwa panzvimbo inosunga uye anosunga semacomplexes.Sezvo iyo mvura inopfuura nepakati peiyo incubation uye yekuona maplateti, iwo macomplexes anotorwa nemamorekuru ekutora ari pabvunzo uye mitsara yekudzora, ichiratidza mhedzisiro inogona kuverengerwa zvakananga kune yakashama ziso.Kazhinji, LFA inogona kupedzwa mumaminitsi 2-15, inokurumidza kupfuura kuwanikwa kwechinyakare.Nekuda kweiyo yakakosha mashini, LFA inoda mashoma ma operation uye haidi yekuwedzera michina, iyo inoita kuti ishandiswe zvakanyanya mushandisi.Zviri nyore kugadzira uye kuita miniaturize, uye mutengo wemapepa-based substrates wakaderera.Zvisinei, inongoshandiswa pakuongorora kwehutano, uye kuongororwa kwehuwandu kwakaoma zvikuru, uye kukwanisa kuwanda kwekuwedzera uye kubuda zvishoma, uye imwe chete yakakwana nucleic acid inogona kuonekwa panguva [96,110,127].
Kunyange zvazvo kushandiswa kwakawanda kweLFA kwakanangidzirwa kune immunoassays, kushandiswa kweLFA kwekuongorora kwema molecular mu microfluidic chips zvakare inoshanda uye yakakurumbira [136].Panyaya yehutachiona hwehepatitis B, HIV uye SARS-CoV-2 LFA Gong et al.[137] yakaronga up-conversion nanoparticle LFA chikuva uye yakaratidza kushandiswa-siyana kweiyi miniaturized uye inotakurika chikuva kuburikidza nekuziva uye kuwanda kwekuona kwezvakawanda zvinangwa seHBV nucleic acid.Mukuwedzera, Fu et al.[138] yakaratidza novel LFA yakavakirwa pamusoro-inosimudzira Raman spectroscopy yehuwandu hwekuongororwa kweHIV-1 DNA pakadzika.Nekukurumidza uye kunzwisiswa kwekuonekwa kweSARS-CoV-2, Liu et al.[85] yakagadzira microfluidic-yakabatanidzwa RPA lateral flow analysis nekubatanidza RT-RPA uye universal lateral flow monitoring system mune imwechete microfluidic system.
Kushandiswa kwemapuratifomu akasiyana emicrofluidic anosiyana zvichienderana nezvidzidzo chaizvo, kutora mukana wakazara wekwaniso uye zvakanakira mapuratifomu.Iine mavharuvhu anokwanisika, pombi uye madhaidhi, LOCC ndiyo yakanyanya kuzara chikuva chekusiyana kwekushandisa uye kudyidzana nekamuri huru yekusimudzira.Naizvozvo, tinovimba uye tinokurudzira kuti zvidzidzo zvitsva zviitwe kuLOCC seyekutanga kuedza uye kuti mamiriro acho agadziriswe.Pamusoro pezvo, nzira dzakanyatsoshanda uye dzakarurama dzinotarisirwa kuwanikwa uye kushandiswa muhurongwa.LOAD inokwenenzvera mukudzora chaiko kwezvinwiwa kubva kumidziyo iripo yeLOCC uye inoratidza zvakanakira dhiraivha imwe chete necentrifugal simba pasina kudiwa kwemadhiraivha ekunze, nepo mhinduro dzakafanana dzinogona kupatsanurwa uye kuwiriraniswa.Nekudaro, mune ramangwana, LOAD ichava iyo huru microfluidic chikuva ine mashoma manyorerwo mashandiro uye akakura uye otomatiki matekinoroji.Iyo µPAD chikuva inosanganisa mabhenefiti eLOCC uye mapepa akavakirwa zvinhu zvemutengo wakaderera, kushandiswa kumwe chete kuongororwa.Naizvozvo, kusimudzira kweramangwana kunofanirwa kutarisisa kune ari nyore uye akasimbiswa matekinoroji.Pamusoro pezvo, iyo LFA yakanyatso kuenderana nekuona maziso asina kupfeka, ichivimbisa kuderedza kushandiswa kwemuenzaniso uye kukurumidza kuona.Kuenzanisa kwakadzama kwepuratifomu kunoratidzwa muTafura 2.
Digital inoongorora inokamura sampu mune akawanda mamicroreactors, imwe neimwe ine discrete nhamba yemamorekuru anotangwa [139, 140].Digital assays inopa mabhenefiti akakosha ekuita huwandu hwakakwana nekuita zviuru zvezvakafanana biochemical kuyedza panguva imwe chete uye mumwe nemumwe mu micron scale compartments kwete muchikamu chinoenderera.Kuenzaniswa neyakajairwa microfluidics, kuita kwekamuri kunogona kuderedza vhoriyamu yemuenzaniso, kuwedzera maitiro ekuita, uye kubatanidzwa nyore nyore nedzimwe nzira dzekuongorora pasina kudiwa kwemachaneli, pombi, mavharuvhu, uye compact dhizaini [141, 142, 143, 144, 145, 146, 147] .Nzira mbiri dzinotevera dzinoshandiswa mumadhijitari assays kuti awane yunifomu uye yakarurama kupatsanurwa kwezvigadziriso, kusanganisira reagents uye sampuli dzakadai semasero, nucleic acids, uye mamwe mapeji kana mamorekuru: (1) inodonha emulsions inoshandisa kusagadzikana kwemvura inoshandiswa;(2) array division inoitwa neiyo geometric constraints yemudziyo.Munzira yekutanga, madonhwe ane reagents uye samples mune microchannels anogona kugadzirwa nenzira dzisiri dzekuita senge co-current, crossflow, kuyerera kwekutarisa, staged emulsification, microchannel emulsification, uye membranes kuburikidza viscous shear masimba uye emulsification ine chiteshi shanduko.localization [143, 145, 146, 148, 149] kana kushandisa nzira dzinoshanda [150, 151], iyo inotanga kuwedzera simba kuburikidza nemagetsi, magnetic, thermal uye mechanical control.Munzira yekupedzisira, iyo yakanakisa fluid volume inoenderana mumakamuri emicrofluidic inogovaniswa nekuchengeta zvimiro zvemukati zvehukuru hwakafanana, senge micropits uye pamusoro arrays [152,153,154].Zvikuru, madonhwe ndiwo makuru ekuyerera zvikamu izvo zvinogona zvakare kugadzirwa uye kushandiswa pamagetsi emagetsi anoenderana nedhijitari microfluidics (DMF).Electrowetting of dielectrics ndeimwe yeakanyanya kudzidzwa DMF theories, sezvo electrowetting ye dielectrics inobvumira kunyatso gadzirisa madonhwe ega, kutonga chimiro chemvura uye asymmetric magetsi masaini anopfuura nemativi akasiyana [141, 144].Mabasa makuru ane madonhwe muDMF anosanganisira kuronga, kupatsanura, nekubatanidza [151, 155, 156], iyo inogona kushandiswa munzvimbo dzakasiyana-siyana dzekuongorora, kunyanya mukuonekwa kwema molecular [157, 158, 159].
Digital nucleic acid yekuona ndeyechitatu-chizvarwa molecular diagnostic tekinoroji inotevera yakajairwa PCR uye quantitative chaiyo-nguva PCR (qPCR), inofambirana nepamusoro-kuburikidza kutevedzana uye mvura biopsy.Mumakumi maviri emakore apfuura, digital nucleic acids yakakurumidza kugadzirwa mumunda wekuongororwa kwema molecular ezvirwere zvinotapukira [160, 161, 162].Mhedziso yehuwandu hwedhijitari nucleic acid yekuona inotanga nekurongedza masampuli uye reagents mumakamuri ega ega kuti ive nechokwadi chekuti kutevedzana kwega kwega kune mukana wakafanana wekupinda mukamuri yega yega.Tichifunga, chikamu chega chega chinogona kupihwa akawanda anoteedzana, kana panogona kunge pasina yakazvimirira microreaction system.Kuburikidza nemaitiro akasiyana ekunzwa atsanangurwa pamusoro, macompartments ane microbial target sequences ayo anoburitsa masaini pamusoro pechimwe chikumbaridzo anogona kuoneswa neziso rakashama kana nemuchina uye akanyorwa seakanaka, nepo mamwe macompartments anogadzira masaini pasi pechikumbaridzo akanyorwa seakanaka. .zvisina kunaka, izvo zvinoita kuti chiratidzo chechikamu chimwe nechimwe ive boolean.Nekudaro, nekuverenga huwandu hwemakamuri akagadzirwa uye chiyero chezvakanaka zvabuda mushure mekuita, makopi ekutanga emasampuli ebvunzo anogona kufananidzwa uchishandisa Poisson yekugovera fomula pasina kudiwa kweiyo curve yakajairwa, inodiwa pakuongorora kwehuwandu hwemazuva ese. se qPCR.[163] Kuenzaniswa nemaitiro echinyakare mamorekuru ekuongorora nzira, dijitari nucleic acid yekuona ine yakakwira dhigirii ye automation, yepamusoro yekuongorora nekukurumidza nekunzwa, mashoma reagents, kushomeka kusvibiswa, uye nyore kugadzira nekugadzira.Nezvikonzero izvi, kushandiswa kwemadhijitari assays, kunyanya kudonhedza-yakavakirwa nzira, dzekuongorora mamorekuru, kusanganisa kukwidziridza uye masaini ekuverenga matekiniki, kwakanyatso kudzidzwa panguva yekubuda kwakasimba kweSARS-CoV-2.Somuenzaniso, Yin et al.[164] yakasanganiswa madonhwe edhijitari uye nekukurumidza PCR nzira dzekuona iyo ORF1ab, N, uye RNase P majini muSARS-CoV-2 mune microfluidic chip.Zvinonyanya kukosha, chirongwa chacho chakakwanisa kuziva chiratidzo chakanaka mukati memasekondi e115, iyo inokurumidza kudarika PCR yakajairwa, ichiratidza kushanda kwayo mukutsvaga-kutarisira (Mufananidzo 7a).Dong et al.[165], Dyara et al.[157], Chen nevamwe.[166] uye Alteri et al.[167] yakashandisawo droplet digital PCR (ddPCR) kuona SARS-CoV-2 mune microfluidic system ine mhedzisiro inonakidza.Kuti uwedzere kuvandudza mwero wekuona, Shen et al.[168] yakawana ddPCR-based chip imaging munguva shoma se15 s pasina kushandisa nzira dzekusona mifananidzo, ichimhanyisa ddPCR tekinoroji maitiro kubva murabhu kuenda kune application.Kwete chete nzira dzekusimudza kupisa dzakadai sePCR dzinoshandiswa, asiwo nzira dzekusimudza isothermal dzinoshandiswa kurerutsa maitiro ekuita uye mhinduro nekukurumidza.Lu et al.[71] yakagadzira SlipChip yekuongorora madonhwe, inokwanisa kuburitsa madonhwe ehukuru hwakasiyana pahupamhi hwepamusoro mune imwe nhanho uye quantifying SARS-CoV-2 nucleic acids uchishandisa dijitari LAMP (Mufananidzo 7b).Seye tekinoroji inokurumidza kubuda, CRISPR inogona zvakare kuita basa rakakosha mudhijitari nucleic acid yekuona kuburikidza neyakajeka colorimetric imaging pasina kudiwa kwekuwedzera nucleic acid tsvina.Ackerman et al.yakagadzira combinatorial matrix reaction ye multiplex evaluation ye nucleic acids.[158] yakaona 169 mavhairasi ane hukama nevanhu, kusanganisira SARS-CoV-2, mumadonhwe ane CRISPR-Cas13-based nucleic acid yekuona reagents mune microwell assay (Mufananidzo 7c).Uye zvakare, isothermal amplification uye CRISPR tekinoroji inogona kushandiswa mune imwecheteyo system kusanganisa mabhenefiti ezvose zviri zviviri.Park et al.[169] A CRISPR/Cas12a dhijitari assay yakagadzirwa mune yekutengeserana microfluidic chip kuti ionekwe yakabviswa uye inouraya-inouraya SARS-CoV-2 zvichibva pane imwe-nhanho RT-RPA ine ipfupi uye yakakwirira chiratidzo-kumashure-kumashure kuona. chiyero chenguva., yakafara dynamic range uye nani kunzwa (Fig. 7d).Dzimwe tsananguro dzemienzaniso iyi dzinopihwa muTafura 3.
Yakajairika dhijitari papuratifomu ye nucleic acid yekuona.a Inokurumidza digital PCR workflow ine matanho mana anokosha: kugadzirira kwemuenzaniso, kugovera kwekuita musanganiswa, maitiro ekusimudzira, uye chinangwa chehuwandu (yakagadziriswa kubva [164]).b Schematic inoratidza ongororo yeSlipChip madonhwe ekugadzira madonhwe pakusimba kwakanyanya (yakagadziriswa kubva [71]).c CARMEN-Cas workflow diagram13 (yakagadziridzwa kubva [158]).d Mhedziso yekuonekwa kwehutachiona hwedhijitari neCRISPR/Cas mupoto imwe (yakatorwa kubva [169]).W/O mvura-mu-mafuta, polydimethylsiloxane PDMS, PCR polymerase chain reaction, DAQ data collection, PID proportional integral derivative, CARMEN combinatorial matrix reaction for multiplex nucleic acid evaluation, SARS-CoV-2, yakanyanya acute breathing syndrome, coronavirus 2 , RT Kukwidziridzwa kwereverse transcriptase recombinase polymerase-RPA, S/B chiratidzo kumashure