Mekhoa ea setso ea ho hlahloba mafu a tšoaetsanoang e hloka hore ho sebelisoe lisebelisoa tsa benchtop tse sa lokelang ho hlahloba lintlha tsa tlhokomelo (POCT).Emerging microfluidics ke thekenoloji e nyenyane haholo, e ikemetseng, le e kopantsoeng eo e ka bang mokhoa o mong oa mekhoa ea setso bakeng sa tlhahlobo e potlakileng, e theko e tlaase, e nepahetseng ea setšeng.Mekhoa ea ho hlahloba limolek'hule e sebelisoa haholo lisebelisoa tsa microfluidic e le mekhoa e sebetsang ka ho fetisisa ea ho lemoha pathogen.Tlhahlobo ena e akaretsa tsoelo-pele ea morao-rao ea tlhahlobo ea limolek'hule e thehiloeng ho microfluidic ea mafu a tšoaetsanoang ho tsoa ponong ea thuto le ea indasteri.Taba ea pele, re hlalosa ts'ebetso e tloaelehileng ea on-chip ea li-nucleic acid, ho kenyeletsoa sampole ea pele ho kalafo, ho holisa, le ho bala matšoao.Litšobotsi, melemo le melemo ea mefuta e mene ea li-platform tsa microfluidic lia bapisoa.Ka mor'a moo, re tla tšohla ts'ebeliso ea litlhahlobo tsa dijithale bakeng sa quantification e felletseng ea li-nucleic acid.Lisebelisoa tsa tlhahlobo ea limolek'hule tsa khale le tsa morao-rao tsa microfluidic li akaretsoa e le bopaki ba boemo ba hajoale ba mmaraka.Qetellong, re sisinya litaelo tsa nako e tlang bakeng sa tlhahlobo ea microfluidic ea mafu a tšoaetsanoang.
Mafu a tšoaetsanoang a bakoa ke likokoana-hloko tse akarelletsang libaktheria, livaerase le likokoana-hloko tse ajoang lefatšeng ka bophara.Ho fapana le mafu a mang, likokoana-hloko li tšoaetsoa kapele 'me li hasana pakeng tsa batho le liphoofolo tse amohelang liphoofolo ka ente, moea le mecha ea litaba ea metsi [1].Thibelo ea mafu a tšoaetsanoang e bohlokoa joalo ka mokhoa oa bophelo bo botle ba sechaba.Mekhoa e meraro e meholo ea ho loantša mafu a tšoaetsanoang: (1) ho laola mohloli oa tšoaetso;(2) tšitiso ea tsela ea phetiso;(3) tshireletso ya batho ba hlaselehang habonolo.Har'a maqheka a ka sehloohong, taolo ea mohloli oa tšoaetso e nkoa e le leano la bohlokoa ka ho fetisisa ka lebaka la boiketlo ba eona le theko e tlaase.Tlhahlobo e potlakileng, ho itšehla thajana le kalafo ea batho ba tšoaelitsoeng ke ntho ea bohlokoa, e hlokang maano a potlakileng, a hlokolosi le a nepahetseng a tlhahlobo [2].Tlhahlobo ea hajoale ea mafu a tšoaetsanoang hangata e kopanya tlhahlobo ea bongaka e ipapisitseng le matšoao le matšoao le lithuto tsa laboratori tse kang setso sa lisele le tlhahlobo ea limolek'hule, tse hlokang basebetsi ba koetlisitsoeng, lits'ebetso tse matla haholo, le lisebelisoa tsa tlhahlobo tse turang [3, 4].Thibelo ea ho qhoma ha mafu a tšoaetsanoang e hloka tlhahlobo e potlakileng, e theko e tlaase, le e nepahetseng ea sebaka seo, haholo-holo libakeng tse nang le lisebelisoa tse fokolang moo mafu a tšoaetsanoang a tloaelehileng le a matla [5], hammoho le phekolo lefeelleng kapa lebaleng la ntoa, moo maemo a tšohanyetso a sa lebelloang..tlhokomelo ea meriana e lekanyelitsoe [6].Tabeng ena, microfluidics ke thekenoloji e kopanyang theknoloji ea microelectromechanical systems, nanotechnology, kapa mahlale a thepa bakeng sa ho qhekella ka mokhoa o nepahetseng oa metsi [7,8,9,10], ho fana ka menyetla e mecha ea ho lemoha ntlha ea tlhokomelo (POCT).) mafu a tšoaetsanoang ka ntle ho lipetlele le lilaboratori.Ha ho bapisoa le litlhahlobo tsa khale tse jang nako, theknoloji ea microfluidic e fana ka sampole le ho boloka litšenyehelo bakeng sa tlhahlobo ea limolek'hule nakong ea ho phatloha ha mafu.Ts'oaetso ea lefats'e ea lefu la coronavirus 2019 (COVID-19) e bakoa ke lefu le matla la ho hema coronavirus 2 (SARS-CoV-2), kahoo bohlokoa ba microfluidics bakeng sa thibelo le taolo ea seoa ka nako bo boetse bo hatisoa [11, 12. , 13].Ho fapana le tlhahlobo ea setso, POCT ea microfluidic e sebelisa lisebelisoa tse nyane tse nkehang ho tloha ho li-analyzer tsa benchtop ho isa ho li-strip tse nyane tsa liteko tsa sidestream ho etsa liteko haufi le sebaka sa sampole [14].Liteko tsena li na le boitokiso bo bonolo kapa bo se nang lisampole, ho holisa lets'oao ka potlako, le ho bala ha matšoao a bobebe ho bakang nako e khuts'oane le liphetho tse nepahetseng ka mor'a metsotso e seng mekae.Ho fumaneha le tlhahiso e kholo ea lisebelisoa tsa tlhokomelo ea bophelo tse thehiloeng ho microfluidic li ekelitse lits'ebetso tsa bona tsa tlhahlobo tse theko e tlaase le tse tobileng ka ntle ho sepetlele, haufi le mokuli, esita le lapeng.
Har'a mekhoa e teng ea ho hlahloba mafu a tšoaetsanoang, tlhahlobo ea limolek'hule ke e 'ngoe ea tse thata ka ho fetisisa [15, 16].Ntle le moo, tlhahlobo ea limolek'hule e atisa ho sebelisoa e le tekanyetso ea khauta bakeng sa tlhahlobo e tsoelang pele ea COVID-19, e lumellang ho fumanoa ka kotloloho hoa libaka tse khethehileng tsa vaerase tsa RNA kapa DNA pele ho karabelo ea 'mele ea ho itšireletsa mafung [17, 18].Tlhahlobisong ea morao-rao, re fana ka tsoelo-pele ea morao-rao ea mekhoa ea ho hlahloba limolek'hule tse thehiloeng ho microfluidics bakeng sa mafu a tšoaetsanoang, ho tloha ponong ea thuto ho ea ho liindasteri tsa nakong e tlang (setšoantšo sa 1).Re tla qala ka mehato e meraro ea bohlokoa ea ho lemoha ha nucleic acid: on-chip sample pretreatment, nucleic acid amplification, le ho bala matšoao.Ka mor'a moo re ile ra bapisa mefuta e fapaneng ea liforomo tsa microfluidic le sebopeho le ts'ebetso ea tsona, tse bontšang litšobotsi tse ikhethang (matla le bofokoli).Ho hlahlobjoa ha digital nucleic acid ho tsoela pele ho buisanoa le ho fanoa e le mohlala oa theknoloji ea moloko oa boraro bakeng sa palo e felletseng ea limolek'hule tsa pathogen e tšoaetsanoang.Ntle le moo, lisebelisoa tse 'maloa tse tloaelehileng le tsa morao-rao tsa POCT li tla hlahisoa ho bonts'a boemo ba hajoale ba mmaraka oa microfluidic POCT bakeng sa tlhahlobo ea limolek'hule.Hape re tla buisana le ho hlalosa pono ea rona bakeng sa likopo tse tlang.
Li-module tsa microfluidic chips bakeng sa ho lemoha ha nucleic acid li ka aroloa ka mekhahlelo e meraro (sampling, recognition, and signing) ho latela mesebetsi ea bona [19].Har'a li-module tsena, mojule oa sampling haholo o hlokomela sampole lysis le nucleic acid extract.Mojule oa sensor o laola haholo phetoho le ho holisa matšoao a nucleic acid.Mojule oa lipontšo o lemoha lets'oao le fetotsoeng le ho sebetsoa ke mojule oa kutlo.Ho ipapisitsoe le mokhoa oa ho bona li-nucleic acid ho chip, re tla akaretsa li-chips tse fapaneng tse ka hlokomelang mosebetsi oa "input and output".
Mohato oa pele oa ho fumana nucleic acid ke ho ntšoa ha asiti ea nucleic, ke hore, ho arola "nucleic acid" ho tloha sampoleng ea mantlha.Nucleic acid extraction e etsoa ho hloekisa nucleic acid ho tsoa ho litšila tse ling tsa limolek'hule, ho netefatsa botšepehi ba sebopeho sa mantlha sa limolek'hule tsa nucleic acid, le ho ntlafatsa sephetho.Ho ntšoa ha Nucleic acid ho hloka sampole e hlokahalang ea lysis le nucleic acid capture, boleng le katleho ea tsona tse nang le tšusumetso e kholo lipatlisisong le liphethong tsa tlhahlobo.Litla-morao leha e le life tse poteletseng nakong ea ho ntšoa li ka fokotsa ho fumanoa hape.Mohlala, mekhoa ea polymerase chain reaction (PCR) le mekhoa ea loop isothermal amplification (LAMP) e thibetsoe ke li-solvents tse ling tse setseng tse kang ethanol le isopropanol ho li-nucleic acid isolation reagents [20].Ho ntšoa ha metsi-mokelikeli le ho ntšoa ha mekhahlelo e tiileng ke mekhoa e tsebahalang haholo ea ho arola li-nucleic acid [21], leha ho le joalo, ho ntšoa ha metsi a mokelikeli ho chip ho na le moeli haholo, kaha li-reagents tse sebelisoang ho ntšoa ha metsi-mokelikeli li baka ho bola ha li-microfluidic chips. .Mona, re totobatsa mekhoa e tiileng ea ho ntša karolo e thehiloeng ho microarray le ho bapisa melemo le mathata a bona.
Silicon ke thepa ea substrate e tsamaellanang le nucleic acid ka lebaka la biocompatibility ea eona, botsitso, le ho feto-fetoha habonolo ha eona [22].Habohlokoa, ha e fetoloa ka silika kapa lisebelisoa tse ling, motsoako ona o bonts'a thepa ea ho bapatsa li-nucleic acid tse qosoang hampe tlas'a pH e tlase, maemo a letsoai a phahameng ha a ntse a e-na le pH e phahameng, litharollo tse tlase tsa letsoai.Ho itšetlehile ka ketsahalo ena, hoa khoneha ho hloekisa nucleic acid.
Mefuta e fapaneng ea lisebelisoa tse thehiloeng ho silica e 'nile ea sebelisoa bakeng sa ho ntša nucleic acid ka microfluidics, tse kang lifaha tsa silica, powders, microfiber filters, le li-membrane tsa silika [23,24,25,26].Ho itšetlehile ka thepa ea thepa, lisebelisoa tse thehiloeng ka silicon li ka sebelisoa ka microcircuits ka litsela tse sa tšoaneng.Ka mohlala, li-granules tsa silika, li-powders, le li-nanofilters tsa khoebo li ka kenngoa ka har'a li-pores kapa li-microchannel tsa li-microfluidic chips le ho thusa ho ntša nucleic acid ho tloha lisampoleng [27, 28, 29].Li-membrane tsa silica tse fetotsoeng holim'a metsi li ka boela tsa sebelisoa ho hloekisa DNA ka potlako ho likokoana-hloko ka theko e tlaase.Ka mohlala, Wang et al.[30] Ka ho kopanya liketso tsa denaturing amplification le vesicle-mediated chain exchange le silica membranes tse koahetsoeng ka li-chitosan oligosaccharides, ho ile ha hlahisoa tsamaiso e nkehang habonolo e fumaneng ka katleho lihlopha tsa 102-108 tsa ho etsa li-colon.(CFU)/ml Vibrio parahaemolyticus., ’me boteng ba kokoana-hloko bo ne bo bonahala habonolo.Powell le ba bang.[31] Li-microarrays tse thehiloeng ho silicon li ile tsa sebelisoa ho lemoha kokoana-hloko ea hepatitis C (HCV), kokoana-hloko ea immunodeficiency (HIV), kokoana-hloko ea Zika, le papillomavirus ea motho le ho ata ka mokhoa o itekanetseng, moo 1.3 μl tortuous microreactor e ileng ea hlahisoa ho tšoara likokoana-hloko tsa RNA.le ho etsa in situ amplification.Ho phaella mekhoeng ena, li-microcolumn tsa silika tse fetotsoeng holim'a metsi li boetse li phetha karolo ea bohlokoa tabeng ea ho ntša nucleic acid, kaha geometry le thepa ea thepa ea ho fetola li eketsa haholo katleho ea ho ntša.Chen et al.[32] e hlahisitse sethala sa microfluidic bakeng sa ho itšehla thajana ha RNA e tlaase-concentration e thehiloeng ho li-amino-coated silicon microcolumns.Sesebelisoa sena sa microfluidic se kopanya mefuta e mengata ea 0.25 cm2 micropillars holim'a silicon substrate ho finyella katleho e phahameng ea ho ntša ka holim'a sebaka se phahameng ho ea ho moralo oa tekanyo ea molumo.Monyetla oa moralo ona ke hore sesebelisoa sa microfluidic se ka fihlela katleho ea 95% ea nucleic acid.Maano ana a thehiloeng ho silicon a bonts'a boleng ba ho arola li-nucleic acid ka potlako ka theko e tlase.Ha e kopantsoe le li-chips tsa microfluidic, maano a ho ntša silicon a ke ke a eketsa bokhoni ba ho lemoha ha nucleic acid feela, empa hape a thusa ho fokotsa le ho kopanya lisebelisoa tsa tlhahlobo [20].
Mekhoa ea karohano ea makenete e sebelisa likaroloana tsa makenete ho arola li-nucleic acid boteng ba matla a khoheli a kantle.Likaroloana tsa makenete tse sebelisoang ka tloaelo li kenyelletsa Fe3O4 kapa γ-Fe2O3 likaroloana tsa makenete tse koahetsoeng ka silika, amino le carboxyl [33,34,35,36].Tšobotsi e khethollang ea likaroloana tsa makenete ha li bapisoa le mekhoa ea silicon-based SPE ke boiketlo ba ho qhekella le ho laola ka limakete tsa ka ntle.
U sebelisa tšebelisano ea electrostatic lipakeng tsa nucleic acid le silika, tlas'a maemo a letsoai le phahameng le pH e tlase, li-nucleic acid li adsorbed ka holim'a likaroloana tsa khoheli tse koahetsoeng ke silika, ha tlas'a maemo a letsoai le tlase le pH e phahameng, limolek'hule li ka hlatsuoa. hape..Lifaha tsa makenete tse koahetsoeng ka silika li etsa hore ho khonehe ho ntša DNA ho lisampole tse kholo tsa molumo (400 μL) ho sebelisoa motsamao o laoloang ka matla [37].E le pontšo, Rodriguez-Mateos et al.[38] e sebelisitse limakenete tse khonang ho laola phetisetso ea lifaha tsa makenete likamoreng tse fapaneng.Ho ipapisitsoe le likaroloana tsa khoheli tse koahetsoeng ka silica, likopi tse 470/mL tsa SARS-CoV-2 genomic RNA li ka ntšoa lisampoleng tsa metsi a litšila bakeng sa tlhahlobo ea LAMP reverse transcript (RT-LAMP) mme karabo e ka baloa nakong ea hora e le 'ngoe.mahlo a hlobotse (Setšoantšo sa 2a).
Lisebelisoa tse thehiloeng ho lisebelisoa tsa makenete le tsa porous.Setšoantšo sa mohopolo oa sesebelisoa sa microfluidic sa IFAST RT-LAMP bakeng sa ho lemoha SARS-CoV-2 RNA (e nkiloe ho [38]).b Sesebelisoa se senyenyane sa Centrifugal bakeng sa dSPE ea buccal swab nucleic acid (e fetotsoeng ho tloha [39]).c Sekontiri sa sampole se ikahelang ka matla se sebelisa karete ea FTA® (e fetotsoeng ho [50]).d Fusion 5 sefa pampiri e fetotsoeng ka chitosan (e nkiloe ho [51]).SARS-CoV-2 e matla haholo ea ho hema lefu la coronavirus 2, RT-LAMP reverse transcript loop mediated isothermal amplification, balekane ba theknoloji ea FTA finders, NA nucleic acid
Likaroloana tsa makenete tse qosoang hantle li loketse ho hokela mokokotlo oa phosphate oa asiti ea nucleic.Sebakeng se itseng sa letsoai, lihlopha tsa phosphate tse qosoang hampe tsa li-nucleic acid li ka qosoa ka mokhoa o nepahetseng holim'a likaroloana tsa motsoako oa makenete.Ka hona, ho ile ha etsoa li-nanoparticles tsa makenete tse nang le bokaholimo bo thata le bongata bo boholo ba lihlopha tsa amino bakeng sa ho ntšoa ha li-nucleic acid.Kamora ho arohana ha makenete le ho thibela, li-magnetic nanoparticles le DNA complexes li ka sebelisoa ka kotloloho ho PCR, e felisang tlhoko ea ts'ebetso e rarahaneng le e jang nako le ts'ebetso ea elution [35].Li-nanoparticles tsa Magnetic tse koahetsoeng ka lihlopha tse mpe tsa carboxyl le tsona li 'nile tsa sebelisoa ho arola li-nucleic acid tse adsorbed holim'a metsi a mangata a polyethylene glycol le sodium chloride solutions [36].Ka lifaha tsena tsa makenete tse fetotsoeng holim'a metsi, ho ntšoa ha DNA ho tsamaisana le ho holisa ho latelang.Dignan et al.[39] e hlalositse sethala se ikemetseng le se nkehang sa centrifugal microfluidic bakeng sa kalafo ea nucleic acid, e lumellang basebetsi bao e seng litsebi ho e sebelisa setšeng.Ho phaella moo, ho lumellana ha DNA e ka thōko le LAMP, mokhoa o loketseng hantle bakeng sa tlhahlobo ea nucleic acid ea ntlha-of-care, ho tsoela pele ho bontša litlhoko tse fokolang tsa thepa le ho tšoaneleha bakeng sa liteko tsa colorimetric (setšoantšo sa 2b).
Mekhoa ea lifaha tsa Magnetic e fana ka monyetla oa ho ntšoa ka boiketsetso, tse ling tsa tsona li teng ho li-extractor tsa li-nucleic acid tsa khoebo [KingFisher;ThermoFisher (Waltham, MA, USA), QIAcube® HT;CapitalBio (Beijing, Chaena) le Biomek®;Beckman (Miami, USA).), Florida, USA)].Melemo ea ho kopanya lifaha tsa makenete le microfluidics e ka sebelisoa bakeng sa ho ntšoa ka mokhoa o ikemetseng oa li-nucleic acid, tse ka ntlafatsang nts'etsopele ea tlhahlobo ea limolek'hule;leha ho le joalo, motsoako oa lifaha tsa makenete tse nang le microfluidics o ntse o its'etleha haholo ho litsamaiso tse rarahaneng tsa taolo bakeng sa tokiso e nepahetseng ea lifaha tsa makenete, e hlalosang ho tuma ha lihlahisoa tsa khoebo e le tse ngata le tse theko e boima, tse fokotsang ts'ebeliso e eketsehileng ea lifaha tsa makenete ho POCT.
Lisebelisoa tse 'maloa tse nang le li-porous tse kang li-filters tsa nitrocellulose tse fetotsoeng, likarete tsa Finders Technology Associates (FTA), lipampiri tse thehiloeng ho polyethersulfone, le lisebelisoa tse koahetsoeng ka glycan le tsona li sebelisitsoe bakeng sa ho lemoha nucleic acid [40, 41, 42, 43, 44].Lisebelisoa tse nang le li-porous fibrous tse kang pampiri e nang le fiber li ile tsa sebelisoa pele ho arola DNA ka ho hohela limolek'hule tsa DNA tse telelehali ka likhoele.Li-pores tse nyane li lebisa thibelong e matla ea 'mele ea limolek'hule tsa DNA, tse amang ho ntšoa ha DNA hantle.Ka lebaka la boholo bo fapaneng ba pore ea pampiri ea fibrous, katleho ea ho ntša ha e khone ho finyella litlhoko tsa DNA amplification [45, 46].Karete ea FTA ke pampiri ea filthara ea khoebo e sebelisoang lefapheng la bongaka ba forensic mme e sebelisoa haholo libakeng tse ling tsa tlhahlobo ea limolek'hule.Ka ts'ebeliso ea pampiri ea selulose e kentsoeng ka lik'hemik'hale tse fapaneng ho thella lisele tsa lisele tsa sampole, DNA e lokollotsoeng e sirelelitsoe ho senyeha ho fihlela lilemo tse 2.Haufinyane tjena, pampiri ea cellulose e kentsoeng e entsoe bakeng sa ho lemoha limolek'hule tsa likokoana-hloko tse fapaneng, ho kenyeletsoa SARS-CoV-2, leishmaniasis, le malaria [47,48,49].HIV ka har'a plasma e ka thōko e lysed ka ho toba, 'me kokoana-hloko ea nucleic acid e ntlafatsoa ka har'a lesela la FTA® la phallo le hahiloeng ka har'a concentrator, e leng se lumellang tlhahiso e sebetsang ea nucleic acid [50] (Fig. 2c).Bothata bo ka sehloohong ba ho lemoha ha nucleic acid ho sebelisa likarete tsa FTA ke hore lik'hemik'hale tse kang guanidine le isopropanol li thibela liketso tse latelang tsa amplification.Ho rarolla bothata bona, re thehile pampiri ea filthara ea Fusion 5 chitosan-modified, e kopanyang melemo ea ho kopanya limolek'hule tsa DNA le pampiri e sefang ea fibrous, le adsorption ea electrostatic ea DNA ho metsoako e fetotsoeng ea chitosan ho fihlela ts'ebetso e ntle haholo ea nucleic acid. ..likhoele tsa sefa [51] (Setšoantšo sa 2d).Ka ho tšoanang, Zhu et al.[52] e bontšitse mokhoa oa PCR o fetotsoeng ka chitosan o thehiloeng ho in situ capillary microfluidic system bakeng sa ho itšehla thajana ka potlako le ho lemoha Zika kokoana-hloko RNA.Nucleic acids e ka adsorbed / desorbed ka har'a motsoako oa lysate / PCR, ka ho latellana, ho itšetlehile ka thepa ea ho tima / ho tima ea chitosan.on and off”, e arabela pH.
Joalokaha ho boletsoe ka holimo, maqheka ana a kopanya melemo ea lisebelisoa tse fapaneng tsa mekhahlelo e tiileng le ho eketsa katleho ea ho ntšoa ha nucleic acid ho microfluidics.Lits'ebetsong tse sebetsang, tšebeliso ea lisebelisoa tsena ka bongata ha e na moruo, 'me phekolo e nepahetseng ea holim'a metsi kapa ho fetoloa ha holim'a lintho tse tloaelehileng ka lisebelisoa tsena le tsona li ka boloka mosebetsi oa tsona.Ka hona, ho lumeloa hore ts'ebetsong ea mekhoa ena ka mor'a thuto ea lifofane e ka fokotsa litšenyehelo.
Teko ea Nucleic acid ho li-platform tsa microfluidic hangata e sebelisa li-sample tse nyane (<100 µl), ka hona e hloka ho holisoa ha li-nucleic acid tse nang le li-probes tse ikhethileng bakeng sa ho fetolela lets'oao le bonolo bakeng sa ho lemohuoa ka tlase ho noka (optical, motlakase, le makenete) [53, 54]. Teko ea Nucleic acid ho li-platform tsa microfluidic hangata e sebelisa li-sample tse nyane (<100 µl), ka hona e hloka ho holisoa ha li-nucleic acid tse nang le li-probes tse ikhethileng bakeng sa ho fetolela lets'oao le bonolo bakeng sa ho lemohuoa ka tlase ho noka (optical, motlakase, le makenete) [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]. Ha ho lekoa li-nucleic acid ho li-platform tsa microfluidic, hangata ho sebelisoa sampole e nyane (<100 µL), ka hona, ho holisoa ha li-nucleic acid tse nang le li-probes tse khethehileng hoa hlokahala ho e fetola lets'oao le bonolo bakeng sa ho lemohuoa ka mor'a moo (optical, motlakase, le makenete) [53,54].微流控微流控 上 的 检测 μ 使用 使用 使用 使用 使用 小样本量 小样本量 便于 便于 便于 检测 检测 检测 检测 检测) 和 和].微流控 平台 上 的 检测 使用 小样本量 小样本量 小样本量 小样本量, ]. 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]]. Ho sibolloa ha li-nucleic acid ho li-platform tsa microfluidic hangata ho sebelisa sampole e nyane (<100 μl), e hlokang ho holisoa ha li-nucleic acid tse nang le li-probes tse khethehileng ho li fetolela hore e be mats'oao bakeng sa ho bonoa (optical, electrical, and magnetic) [53, 54]] .Nucleic acid amplification ho microfluidics e ka boela ea potlakisa karabelo, ea ntlafatsa meeli ea ho lemoha, ea fokotsa litlhoko tsa sampuli, le ho ntlafatsa ho nepahala ha ho lemoha [55, 56].Lilemong tsa morao tjena, ka temoho ea ho lemoha ka potlako le ka nepo, mekhoa e fapaneng ea ho holisa nucleic acid e sebelisitsoe ho microfluidics, ho kenyeletsoa PCR le karabelo e itseng ea isothermal amplification.Karolo ena e tla akaretsa mekhoa ea ho lemoha ha nucleic acid e ipapisitse le litsamaiso tsa microfluidic.
PCR ke papiso ea ts'ebetso ea phetisetso ea DNA ea ntho e phelang, eo khopolo ea eona e hlalosoang ka botlalo libakeng tse ling 'me e ke ke ea tšohloa mona.PCR e ka holisa palo e nyane haholo ea sepheo sa DNA/RNA ka sekhahla sa exponential, ea etsa PCR sesebelisoa se matla sa ho lemoha ka potlako li-nucleic acid.Lilemong tse mashome tsa morao tjena, lisebelisoa tse ngata tse nkehang tsa microfluidic tse nang le lisebelisoa tsa libaesekele tse futhumatsang tsa PCR li 'nile tsa ntlafatsoa ho finyella litlhoko tsa tlhahlobo ea tlhokomelo ea tlhokomelo [57, 58].PCR ea on-chip e ka aroloa ka mefuta e mene (e tloaelehileng, phallo e tsoelang pele, e fetohang sebaka, le PCR ea convective) ho latela mekhoa e fapaneng ea ho laola mocheso [59].Ka mohlala, Gee et al.[60] e hlahisitse mokhoa o tobileng oa reverse transcription quantitative PCR (RT-qPCR) sethaleng sa bona sa microfluidic bakeng sa ho lemoha multiplex ea SARS-CoV-2, livaerase tsa ntaramane A le B ka lisampole tsa 'metso (Setšoantšo sa 3a) .Park et al.[61] e hahile chip e bonolo ea ho hlahloba likokoana-hloko ka ho kopanya filimi e tšesaane ea PCR, li-electrode, le monoana oa polydimethylsiloxane-based microfluidic module.Leha ho le joalo, mesebetsi ka bobeli e kenyelletsa mefokolo e tloaelehileng ea PCR e tloaelehileng.PCR e hloka ho palama baesekele e futhumetseng, e fokotsang ts'ebetso e nyane ea lisebelisoa le nako e fokotsehileng ea liteko.
Nts'etsopele ea phallo e tsoelang pele e thehiloeng ho microfluidic le PCR e fetotsoeng sebaka e bohlokoa ho rarolla bothata bona.Ho sebelisa mocha o molelele oa noha kapa mocha o khutšoanyane o otlolohileng, PCR e phallang e tsoelang pele e ka fana ka matlafatso e potlakileng ka ho potoloha li-reagents ka mafolofolo libakeng tse tharo tsa preheat ka pompo ea off-chip.Ts'ebetso ena e atleha ho qoba mohato oa phetoho pakeng tsa mocheso o fapaneng oa karabelo 'me kahoo o fokotsa haholo nako ea teko [62] (Setšoantšo sa 3b).Phuputsong e 'ngoe ea Jung et al.[63] e hlahisitse mochine o mocha oa rotary PCR genetic analyzer e kopanyang litšobotsi tsa PCR e tsitsitseng le e phallang bakeng sa ultrafast le multiplex reverse transcription PCR (Fig. 3c).Bakeng sa amplification ea nucleic acid, PCR microchip e tla fetisoa ka li-blocks tse tharo tsa mocheso ka mocheso o fapaneng: 1. Denaturation block 94 ° C, 2. Anealing block ho 58 ° C, 3. Sebaka sa ho atolosa ho 72 ° C.
Tšebeliso ea PCR ho microfluidics.Kemelo ea moralo oa dirRT-qPCR sethaleng sa microfluidic (e nkiloe ho [60]).b Boemeli ba moralo oa phallo e tsoelang pele ea PCR microarray e thehiloeng mocha oa noha (o nkiloe ho [62]).c Pontšo ea moralo oa mohlahlobi oa liphatsa tsa lefutso oa rotary PCR, o nang le microchip, li-block tse tharo tsa ho futhumatsa le mochini oa stepper (o nkiloe ho [63]).d Setšoantšo sa thermoconvection PCR e nang le centrifugation le setup (e nkiloe ho [64]).DirRT-qPCR, e tobileng quantitative reverse transcription polymerase chain reaction
U sebelisa li-capillaries le loops kapa esita le lipoleiti tse tšesaane, PCR ea convection e ka holisa ka potlako li-nucleic acid ka convection ea tlhaho e sa lefelloeng ea mocheso ntle le tlhoko ea pompo e kantle.Ka mohlala, sethala sa cyclic olefin polymer microfluidic se ile sa hlahisoa sethaleng sa ho futhumatsa se pota-potiloeng se sebelisang libaesekele tse futhumatsang ka centrifugation ho PCR loop microchannel [64] (Fig. 3d).Tharollo ea karabelo e tsamaisoa ke convection ea mocheso, e tsoelang pele ho fapanyetsana mocheso o phahameng le o tlaase ho microchannel e nang le mohaho oa annular.Ts'ebetso eohle ea amplification e ka phethoa ka metsotso e 10 ka moeli oa ho lemoha oa 70.5 pg/channel.
Joalo ka ha ho lebelletsoe, PCR e potlakileng ke sesebelisoa se matla sa tlhahlobo ea limolek'hule tse kopantsoeng ka botlalo le litsamaiso tsa tlhahlobo ea multiplex.PCR e potlakileng e fokotsa haholo nako e hlokahalang ho lemoha SARS-CoV-2, e thusang taolong e sebetsang ea seoa sa COVID-19.
PCR e hloka sebaesekele se rarahaneng sa mocheso se sa lokelang POCT.Haufinyane tjena, mekhoa ea ho hōlisa isothermal e sebelisitsoe ho microfluidics, ho kenyelletsa empa e sa felle feela ho LAMP, recombinase polymerase amplification (RPA), le amplification e thehiloeng ho tatellano ea nucleic acid [65,66,67,68].Ka mekhoa ena, li-nucleic acid li holisoa ka mocheso o sa khaotseng, e leng ho nolofalletsang ho thehoa ha lisebelisoa tsa POCT tse theko e tlaase, tse nkehang habonolo bakeng sa tlhahlobo ea limolek'hule.
Liteko tse phahameng tsa LAMP tse thehiloeng ho microfluidics li lumella ho fumanoa hangata ha mafu a tšoaetsanoang [42, 69, 70, 71].Ha e kopane le tsamaiso ea centrifugal microfluidic, LAMP e ka tsoela pele ho thusa ho iketsetsa ha nucleic acid discovery [69, 72, 73, 74, 75].SlipChip ea spin-and-react e ile ea ntlafatsoa bakeng sa ho bona libaktheria tse ngata tse tšoanang ho sebelisa LAMP [76] (Fig. 4a).Ha u sebelisa LAMP e ntlafalitsoeng tekong, tekanyo ea lerata la fluorescence e ne e batla e le makhetlo a 5, 'me moeli oa ho lemoha o fihlile ho likopi tse 7.2 / μl tsa genomic DNA. Ho feta moo, ho ba teng ha likokoana-hloko tse hlano tse tloaelehileng tsa likokoana-hloko tsa tšilo ea lijo, ho akarelletsa le Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis le Vibrio parahaemolyticus, li ile tsa bonoa ho latela mokhoa oa <60 min. Ho feta moo, ho ba teng ha likokoana-hloko tse hlano tse tloaelehileng tsa likokoana-hloko tsa tšilo ea lijo, ho akarelletsa le Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis le Vibrio parahaemolyticus, li ile tsa bonoa ho latela mokhoa oa <60 min.Ho feta moo, boteng ba likokoana-hloko tse hlano tse tloaelehileng tsa baktheria tsa pampitšana ea tšilo ea lijo, ho akarelletsa le Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis le Vibrio parahaemolyticus, li ile tsa bonoa ho sebelisoa mokhoa ona ka nako e ka tlaase ho metsotso e 60.此外, 基于基于 该 在 <60 分钟 内 可 可 可 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病 细菌病此外, 基于基于 在 <60 分钟分钟 内 视化 视化 视化 的 的 的 的 的 的 的 的 的 的 的 的 的 的 的 的 的弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 HIPHo feta moo, boteng ba likokoana-hloko tse hlano tse tloaelehileng tsa baktheria tsa ka mpeng, tse akarelletsang Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvius, le Vibrio parahaemolyticus, li ile tsa bonoa ho sebelisoa mokhoa ona ka nako e ka tlaase ho metsotso e 60.
Melemo ea LAMP ho microfluidics e kenyelletsa, har'a tse ling, karabelo e potlakileng le ho lemoha ha miniaturized.Leha ho le joalo, ka lebaka la mocheso oa karabelo (hoo e ka bang 70 ° C), li-aerosol li hlahisoa ka mokhoa o ke keng oa qojoa nakong ea LAMP, e leng se bakang sekhahla se phahameng sa bohata.Sebopeho sa Assay, moralo oa primer, le taolo ea mocheso le tsona li hloka ho ntlafatsoa bakeng sa LAMP.Ntle le moo, meralo ea li-chip e kenyang ts'ebetso ea ho lemoha liphofu tse ngata ho chip e le 'ngoe e bohlokoa haholo 'me e lokela ho ntlafatsoa.Ho phaella moo, LAMP e loketse ho lemoha mekhoa e mengata e kopantsoeng ka har'a chip e le 'ngoe, e leng ea bohlokoa haholo, empa ho ntse ho e-na le sebaka se seholo sa tsoelo-pele.
Sekhahla se phahameng se fosahetseng sa LAMP se ka fokotsoa ka karolo e itseng ka RPA, kaha mocheso o batlang o le tlase (~ 37 ° C) o fella ka mathata a seng makae a mouoane [77].Tsamaisong ea RPA, li-primers tse peli tse fapaneng li qala DNA synthesis ka ho tlama ho recombinase le amplification e ka phethoa nakong ea metsotso ea 10 [78,79,80,81].Ka hona, ts'ebetso eohle ea RPA e potlakile haholo ho feta PCR kapa LAMP.Lilemong tsa morao tjena, theknoloji ea microfluidic e bontšitsoe ho tsoela pele ho ntlafatsa lebelo le ho nepahala ha RPA [82,83,84].Ka mohlala, Liu et al.[85] e hlahisitse tlhahlobo ea microfluidic e kopantsoeng ea lateral flow polymerase recombinase amplification bakeng sa ho lemoha kapele le ka hloko ha SARS-CoV-2 ka ho kopanya reverse transcription RPA (RT-RPA) le universal lateral flow test strip system.ho kena tsamaisong e le 'ngoe ea microfluidic.Setšoantšo sa 4b).Moeli oa ho fumana ke kopi e le 1/µl kapa 30 likopi/sampole, 'me ho fumanoa ho ka phetheloa ka mor'a metsotso e ka bang 30.Kong et al.ba entse sesebelisoa sa microfluidic se ka aparoang.[86] o sebelisitse mocheso oa 'mele le mokhoa oa ho lemoha oa fluorescence oa selefouno ho lemoha ka potlako le ka ho toba DNA ea HIV-1 ho sebelisa RPA (Figure 4c).Teko ea RPA e ka aparoang e fumana likopi tse 100/mL ea tatellano e reriloeng nakong ea metsotso e 24, e bonts'a monyetla o moholo oa tlhahlobo e potlakileng ea masea a nang le tšoaetso ea HIV-1 maemong a fokolang a lisebelisoa.
Isothermal amplification in point-of-care testing (POCT).Ntlafatso le tlhahiso ea spin le reaction SlipChip.Ka mor'a ho cheselletsa plasma, li-chips tse ka holimo le tse ka tlaase li ile tsa bokelloa ka sete ea linate ho etsa chip ea ho qetela (e nkiloe ho [76]).b Schematic ea sistimi ea MI-IF-RPA bakeng sa ho lemoha COVID-19 (e nkiloe ho [85]).c Schematic ea teko ea RPA e ka roaloang bakeng sa ho lemoha kapele HIV-1 DNA (e nkiloe ho [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 light emitting diode, MI-flubites-IF-IF-RPA Recombinase Emitting diode ea LED, Microflubite-IF-IF-RPA In. Matlafatso
RPA e thehiloeng ho Microfluidic e ntse e tsoela pele ka potlako, leha ho le joalo, litšenyehelo tsa ho etsoa ha chip le tšebeliso ea karabelo li holimo haholo 'me li tlameha ho fokotsoa ho eketsa ho fumaneha ha theknoloji ena.Ho feta moo, maikutlo a phahameng a RPA a ka ama ho holisoa ha lihlahisoa tse sa ikhethang, haholo-holo ha ho na le tšilafalo.Meeli ena e ka ama ts'ebeliso ea RPA lits'ebetsong tsa microfluidic mme e tšoaneloa ke ntlafatso e eketsehileng.Li-primers le li-probes tse hlophisitsoeng hantle bakeng sa lipheo tse fapaneng le tsona lia hlokahala ho ntlafatsa bokhoni ba maano a microfluidic a thehiloeng ho RPA ho POCT.
Cas13 le Cas12a li na le bokhoni ba ho arola li-nucleic acid ka mokhoa o sa reroang 'me kahoo li ka ntlafatsoa e le lisebelisoa tsa ho lemoha le ho hlahloba.Cas13 le Cas12a li kentsoe tšebetsong ha li tlamella ho shebisana DNA kapa RNA, ka ho latellana.Ha e se e kentsoe tšebetsong, protheine e qala ho petsola li-nucleic acid tse ling tse haufi, ka mor'a moo li-RNA tse lebisang ho pathogen-specific nucleic acids li ka phunya li-fluorescent probes tse tingoeng ebe li lokolla fluorescence.E itšetlehile ka khopolo ena, Kellner et al.[87] e hlahisitse mokhoa o thehiloeng ho Cas13 [Specific High-sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK)], le Broughton et al.[88] e hlahisitse mokhoa o mong o thehiloeng ho Cas12a [CRISPR Trans Reporter e lebisitseng DNA endonuclease (DTECR)].
Lilemong tsa morao tjena, ho hlahile mekhoa e fapaneng ea ho fumana li-nucleic acid tse thehiloeng ho CRISPR [89, 90].Mekhoa e tloaelehileng e thehiloeng ho CRISPR hangata e ja nako le mosebetsi o boima ka lebaka la mekhoa e mengata e kenyeletsang nucleic acid extract, amplification le CRISPR ho lemoha.Ho pepesetsoa lino tse tahang moeeng ho ka eketsa monyetla oa ho ba le liphetho tse fosahetseng.Ho latela se boletsoeng ka holimo, litsamaiso tse thehiloeng ho CRISPR li hloka haholo ho ntlafatsoa.
Sethala sa microfluidic se laoloang ke moea se ka etsang litlhahlobo tse 24 ka ho ts'oana se ntlafalitsoe bakeng sa lits'ebetso tsa CRISPR-Cas12a le CRISPR-Cas13a [91].Sistimi ena e na le sesebelisoa sa ho lemoha ha fluorescence se fetang amplification ea nucleic acid mme se iphumanela disampole tsa femtomolar DNA le RNA.Chen et al.[92] recombinase amplification e kopantsoeng le tsamaiso ea CRISPR-Cas12a ka centrifugal microfluidics (Setšoantšo sa 5a).Mosebetsi ona o hlola bothata ba ho kopanya lits'ebetso tsena tse peli hobane Cas12a e khona ho cheka DNA ea lenģosa le ho thibela mokhoa oa ho holisa.Ho phaella moo, Chen et al.[92] ho feta moo e bolokile li-reagents tsa karabelo ho centrifugal microfluidic control ho phethela ts'ebetso eohle ka bo eona.Mosebetsing o mong, Silva et al.[93] e hlahisitse mokhoa oa ho hlahloba ntle le ho phahamisa CRISPR / Cas12a le smartphone ho lemoha SARS-CoV-2 (Fig. 5b).Teko ena, e tsejoang ka hore ke sesebelisoa sa mahala sa amplification-based cell phone, se kenyelletsa enzyme e itšetlehileng ka CRISPR / Cas e ipapisitseng le pono ea smartphone ea matšoao a bubble a hlahisoang ke catalase liteisheneng tsa microfluidic.Ho lemoha ka tlase ho likopi tse 50/µl tsa nucleic acid ntle le ho holisa pele, ts'ebetso eohle ho tloha ho ente ea sampole ho ea ho ho bala matšoao ho nka metsotso e 71 feela.
Mekhoa ea ho lemoha ha Nucleic acid e thehiloeng ho CRISPR.Centrifugal POCT bakeng sa tlhahlobo e kopaneng ea limolek'hule e thehiloeng ho CRISPR (e nkiloe ho [92]).b Nts'etsopele ea tlhahlobo ea CASCADE bakeng sa tlhahlobo e thehiloeng ho smartphone ea SARS-CoV-2 (e nkiloe ho [93]).RAA recombinase amplification, PAM e haufi le protospacer motif, CRISPR e kopantsoeng ho pheta-pheta ha nako e khutšoanyane ea palindromic ka nako e tloaelehileng, tsamaiso ea CASCADE ntle le ho phahamisa mohala oa selefouno ka li-enzyme tse itšetlehileng ka CRISPR / CAS, 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride EDC
E le mohato oa ho qetela oa ho lemoha ha nucleic acid, ho lemoha matšoao ho bonahatsa ka ho toba liphetho tsa tlhahlobo 'me ke ntlha ea bohlokoa ho nts'etsopele ea POCT e sebetsang hantle, e hlokolosi le e nepahetseng.Lipontšo li ka baloa ho sebelisoa mekhoa e fapaneng joalo ka maano a fluorescent, electrochemical, colorimetric le makenete.Karolong ena, re hlalosa lebaka la mokhoa o mong le o mong le ho bapisa tlhahlobo ea limolek'hule tsa mafu a tšoaetsanoang ka microfluidics.
Maano a thehiloeng ho Fluorescence a sebelisoa haholo bakeng sa tlhahlobo ea POCT ea mafu a tšoaetsanoang ka lebaka la melemo ea bona e makatsang ea kutlo e ntle haholo, litšenyehelo tse tlase, ts'ebetso e bonolo, le tlhahlobo ea tlhokomelo [94, 95].Maano ana a sebelisa li-fluorophore tse ngotsoeng joalo ka lidae tsa fluorescent le li-nanomaterials ho theha lets'oao le lemohuoang (matlafatso a fluorescence kapa quenching).Sephetho sena se fana ka maikutlo a hore maano a thehiloeng ho fluorescence a ka aroloa ka ho ngola ka ho toba ka fluorescent, ho bonesa le ho lemoha ha fluorescent e tima [96].Ho lemoha li-label tsa fluorescent ka ho toba ho sebelisa li-labels tse khethehileng tsa fluorescent ho tšoaea li-ligands tse itseng tse hlahisang palo e itseng ea fluorescence ha li tlameletsoe ho sepheo.Bakeng sa ho lemoha ha lets'oao la fluorescence, boleng ba lets'oao la fluorescent bo amana hantle le boholo ba thahasello.Matla a Fluorescence ha a na thuso ha ho se na sepheo 'me a bonoa ha palo e lekaneng ea sepheo e le teng.Ka lehlakoreng le leng, matla a fluorescence a bonoang ke "signal-off" fluorescence a bapisoa le palo ea sepheo, qalong e fihla ho boleng bo phahameng 'me butle-butle ea fokotseha ha sepheo se ntse se atolosoa.Ka mohlala, ho sebelisa mochine oa trans-cleavage o itšetlehileng ka sepheo sa CRISPR-Cas13a, Tian et al.[97] e hlahisitse leano la temoho ea novel ho lemoha li-RNA tse fetang ka ho toba (Setšoantšo sa 6a).Ha e tlama li-RNA tse tlatselletsang, mochini oa CRISPR-Cas13-RNA o ka kengoa ts'ebetsong, oa etsa hore ho be le li-transcollateral cleavage ke baqolotsi ba litaba ba seng ba itseng.Motlalehi ea ngotsoeng ka fluorescently [fluorophore (F)] e tingoa ke quencher (Q) e tsitsitseng le fluoresces ha e phunyeletsoa ke motsoako o kentsoeng.
Molemo oa ho lemoha electrochemical ke lebelo le phahameng la ho lemoha, tlhahiso e bonolo, theko e tlaase, e bonolo ho e jara le ho laola ka mokhoa o itekanetseng.Ke mokhoa o matla oa tlhahlobo bakeng sa lits'ebetso tsa POCT.E ipapisitse le li-transistors tsa graphene field-effect Gao et al.[98] e ile ea hlahisa nanobiosensor bakeng sa ho fumana multiplex ea li-antigen tsa lefu la Lyme ho tloha libaktheria tsa Borrelia burgdorferi tse nang le moeli oa ho lemoha oa 2 pg / mL (Fig. 6b).
Litlhahlobo tsa colorimetric li sebelisitsoe lits'ebetsong tsa POCT, li rua molemo melemong ea ho nkeha habonolo, litšenyehelo tse tlase, ho itokisa habonolo, le ho bala ka pono.Ho lemoha colorimetric ho ka sebelisa oxidation ea peroxidase kapa peroxidase-like nanomaterials, aggregation ea nanomaterials, le tlatsetso ea dae e bonts'ang ho fetolela tlhahisoleseling mabapi le boteng ba li-nucleic acid tse hlabollang hore e be liphetoho tse bonahalang tsa mebala [99, 100, 101].Haholo-holo, li-nanoparticles tsa khauta li sebelisoa haholo ho nts'etsopele ea maano a colorimetric, 'me ka lebaka la bokhoni ba tsona ba ho etsa hore ho be le liphetoho tse potlakileng tsa mebala, ho na le thahasello e ntseng e eketseha ho nts'etsopele ea POCT colorimetric platforms bakeng sa ho hlahloba mafu a tšoaetsanoang ka har'a situ [102].Ka sesebelisoa se kopantsoeng sa centrifugal microfluidic [103], likokoana-hloko tse jeoang ke lijo ka lisampole tsa lebese tse silafetseng li ka fumanoa ka tsela e iketsang boemong ba lisele tsa baktheria tsa 10, 'me liphello li ka baloa ka pono nakong ea metsotso ea 65 (Setšoantšo sa 6c).
Mekhoa ea ho lemoha makenete e ka lemoha ka nepo bahlahlobisisi ba sebelisa lisebelisoa tsa makenete, 'me ho bile le thahasello e kholo lits'ebetsong tsa POCT lilemong tse mashome tsa morao tjena.Mekhoa ea ho lemoha ka matla a makenete e na le melemo e ikhethang joalo ka lisebelisoa tsa makenete tse theko e tlase ho fapana le lisebelisoa tse theko e phahameng tsa optical.Leha ho le joalo, tšebeliso ea matla a khoheli e ntlafatsa ts'ebetso ea ho lemoha le ho fokotsa nako ea ho itokisetsa sampole [104].Ntle le moo, liphetho tsa tlhahlobo ea matla a khoheli li bonts'a boits'oaro bo phahameng, kutlo, le karo-karolelano e phahameng ea lerata-ho-lerata ka lebaka la lets'oao le sa reng letho la lisampole tsa baeloji [105].Sharma et al.e kenyellelitse "magnetic tunnel junction" e thehiloeng ho biosensor ho sethaleng sa microchip se nkehang.[106] bakeng sa ho lemoha li-multiplex tsa likokoana-hloko (Setšoantšo sa 6d).Li-biosensors li lemoha ka hloko li-subnanomolar nucleic acid tse arohaneng le likokoana-hloko.
Mokhoa o tloaelehileng oa ho lemoha matšoao.Khopolo ea ho lemoha hyperlocalized ea Cas13a (e fetotsoe ho tloha [97]).b Graphene nanobiosensor FET hammoho le Lyme GroES scFv (e nkiloe ho [98]).c Lits'oants'o tsa colorimetric bakeng sa ho lemoha li-multiplex tsa likokoana-hloko tse jang lijo ka har'a chip ea centrifugal microfluidic: No. 1 le No. 3 lisampole tse nang le likokoana-hloko tse lebisitsoeng, le No. .d Biosensor e thehiloeng holim'a kotopo ea makenete, ho kenyeletsoa sethala, amplifier e thibelang e hahelletsoeng ka hare, yuniti ea taolo, le phepelo ea motlakase bakeng sa tlhahiso ea matšoao / ho fumana (e nkiloe ho [106]).GFET Graphene FET, Escherichia coli, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus, Vibrio parahaemolyticus, Listeria monocytogenes, PC PC, PDMS Dimethicone, PMMA polymethyl methacrylate
Ho sa tsotellehe litšobotsi tse babatsehang tsa mekhoa e ka holimo ea ho lemoha, e ntse e e-na le mefokolo.Mekhoa ena e bapisoa (tafole ea 1), ho kenyeletsoa lits'ebetso tse ling tse nang le lintlha (mehle le likotsi).
Ka nts'etsopele ea microfluidics, microelectromechanical systems, nanotechnology le saense ea thepa, tšebeliso ea lisebelisoa tsa microfluidic bakeng sa ho lemoha mafu a tšoaetsanoang e ntse e tsoela pele [55,96,107,108].Ho qhekella ka nepo ha lisebelisoa tse nyane le maro ho thusa ho nepahala ha tlhahlobo le ho boloka litšenyehelo.Ka hona, bakeng sa nts'etsopele e eketsehileng, ho entsoe boiteko ba ho ntlafatsa le ho ntlafatsa li-chips, ho fella ka li-chips tse fapaneng tsa microfluidic tse nang le meaho le mesebetsi e fapaneng.Mona re kenyelletsa ka bokhutšoanyane mefuta e mengata e tloaelehileng ea li-platform tsa microfluidic le ho bapisa litšobotsi tsa bona (mehle le boiketlo).Ntle le moo, boholo ba mehlala e thathamisitsoeng ka tlase e shebana haholo le ho loants'a SARS-CoV-2.
Li-LOCC ke mekhoa e tloaelehileng ka ho fetisisa ea ho hlahloba e rarahaneng ea miniaturized 'me ts'ebetso ea bona e fokotsehile haholo, e kopantsoe, e iketselitse ebile e bapisoa le ente ea sampole le ho itokisa, taolo ea phallo le ho lemoha metsi [109, 110].Mekelikeli e sebelisoa ka mokhoa o hlophisitsoeng oa geometry le tšebelisano ea litlamorao tse ngata tsa 'mele joalo ka likhatello tsa khatello, ketso ea capillary, electrodynamics, matla a khoheli le maqhubu a acoustic [111].LOCC e bonts'a melemo e babatsehang ea ho hlahloba lintlha tse phahameng le ho lemoha hangata, ka lebelo la ho hlahloba ka potlako, boholo ba sampuli e nyenyane, tšebeliso e tlaase ea matla, le tsamaiso e phahameng le ts'ebetso ea ts'ebetso;leha ho le joalo, lisebelisoa tsa LOCC li bobebe haholo, 'me li etsa, li paka, ebile lia kopana.Leha ho le joalo, ho pheta-pheta le ho sebelisa hape ho tobana le mathata a maholo [96].Ha ho bapisoa le lipolanete tse ling, LOCC e na le melemo e ikhethang mabapi le mefuta e mengata ea ts'ebeliso le tšebelisano e ntle ka ho fetisisa ea thekenoloji, empa mefokolo ea eona e totobetse, e leng ho rarahana ho hoholo le ho se phetehe hantle.Ho itšetleha ka lipompo tsa ka ntle, tse atisang ho ba tse ngata le tse theko e boima, ho fokotsa ho sebelisoa ha tsona ho POCT.
Nakong ea seoa sa COVID-19, LOCC e ile ea fumana tlhokomelo e ngata.Ka nako e ts'oanang, ho na le li-chips tse 'maloa tse ncha tse kopanyang mahlale a mangata.Mohlala, li-smartphones li se li sebelisoa haholo joalo ka lisebelisoa tsa analytics tse nkehang habonolo 'me li na le monyetla o moholo oa ho kopanya LOCC.Letsatsi le al.[21] e entse "microfluidic chip" e lumellang hore ho be le li-multiplexing tse khethehileng tsa nucleic acid tsa likokoana-hloko tse hlano, ho akarelletsa le SARS-CoV-2, ho sebelisa LAMP le ho li hlahloba ka ho sebelisa smartphone nakong ea hora e le 'ngoe ka mor'a ho fela ha karabelo.E le mohlala o mong, Sundah et al.[112] e thehile switch ea limolek'hule [catalytic amplification by molecular transition state switch (CATCH)] bakeng sa ho lemoha ka ho toba le ka hloko lipehelo tsa SARS-CoV-2 RNA ho sebelisa li-smartphone. CATCH e tsamaisana le LOCC e nkehang habonolo mme e fihlela ts'ebetso e phahameng (hoo e ka bang likopi tse 8 tsa RNA / μl; <1 h mocheso oa kamore) [112]. CATCH e tsamaisana le LOCC e nkehang habonolo mme e fihlela ts'ebetso e phahameng (hoo e ka bang likopi tse 8 tsa RNA / μl; <1 h mocheso oa kamore) [112]. CATCH совместим с портативным LOCC и обеспечивает превосходную производительность (примерно 8 копий РНК/мкл; < 1 ч при компечи12] . CATCH e tsamaisana le LOCC e nkehang habobebe 'me e fana ka tlhahiso e ntle haholo (hoo e ka bang likopi tse 8 tsa RNA/µl; <1 h mocheso oa kamore) [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 комнат1т1 тем). CATCH e tsamaisana le li-LOCC tse nkehang habobebe 'me e na le ts'ebetso e ntle haholo (hoo e ka bang likopi tse 8 tsa RNA/µl; <hora e le 1 mochesong oa kamore) [112].Ntle le moo, lisebelisoa tsa LOCC bakeng sa tlhahlobo ea limolek'hule le tsona li sebelisa matla a mang a ho khanna joalo ka vacuum, otlolla le masimo a motlakase.Kang et al.[113] e bonts'itse PCR ea nako ea 'nete, e potlakileng haholo ea nanoplasma-on-a-chip bakeng sa tlhahlobo e potlakileng le e ngata ea COVID-19 lebaleng e sebelisa chip ea PCR ea vacuum plasmonic liquid.Li et al.[114] ka mor'a moo o ile a hlahisa chip ea microfluidic e otlolohileng e ileng ea nolofalletsa ho fumanoa ha COVID-19.Sethala se sebelisa sistimi ea amplification ea RT-LAMP ho fumana hore na sampole e ntle kapa e mpe.Ka mor'a moo, Ramachandran et al.[115] e fihletse li-gradients tsa motlakase tse loketseng ho sebelisa isotachophoresis (ITP), mokhoa o ikhethileng oa ho tsepamisa mohopolo o kentsoeng tšebetsong ho microfluidics.Ka ITP, sepheo sa RNA ho tsoa ho lisampole tse tala tsa nasopharyngeal swab li ka hloekisoa ka bohona.Joale Ramachandran et al.[115] Ho kopanya tlhoekiso ena ea ITP le litlhahlobo tse ntlafalitsoeng tsa LAMP ea ITP le CRISPR ho lemohile SARS-CoV-2 ho swab ea nasopharyngeal ea motho le mehlala ea bongaka ka nako e ka bang metsotso e 35.Ho phaella moo, maikutlo a macha a lula a hlaha.Jadhav et al.[116] e sisintse leano la tlhahlobo le thehiloeng holim'a sebopeho sa Raman se ntlafalitsoeng se kopantsoeng le sesebelisoa sa microfluidic se nang le li-nanotubes tsa khauta tse tšekaletseng / silevera kapa li-electrospun micro/nanotubes tse lahliloeng.Membrane-functionalized ahel-in filter microchannels li ka lahloa.Sesebelisoa se bapisa livaerase tse tsoang marong a fapaneng a 'mele / li-exudations tse kang mathe, nasopharynx le meokho.Kahoo, tšoaetso ea kokoana-hloko e ngata 'me kokoana-hloko e ka tsejoa ka nepo ka ho saena ha Raman.
LOAD ke sethala sa microfluidic sa centrifugal moo lits'ebetso tsohle li laoloang ke protocol ea frequency e potolohang substrate ea microstructured [110].Sesebelisoa sa LOAD se khetholloa ka ho sebelisa matla a centrifugal e le matla a bohlokoa a ho khanna.Mekelikeli e boetse e tlas'a matla a capillary, Euler le Coriolis.U sebelisa sesebelisoa sa centrifuge, litlhahlobo li etsoa ts'ebetsong e tsoelang pele ea mokelikeli ho tloha ho radial ho ea sebakeng sa kantle, ho felisa tlhoko ea li-tubing tse ling tsa kantle, lipompo, li-actuator le li-valve tse sebetsang.Ka bokhutšoanyane, mokhoa o le mong oa ho laola o nolofatsa ts'ebetso.Matla a sebetsang holim'a mokelikeli ka mocha o tšoanang oa microfluidic sebakeng se le seng ho tloha setsing sa mojaro a lekana, e leng se etsang hore ho khonehe ho pheta sebopeho sa mocha.Ka hona, lisebelisoa tsa LOAD li bonolo ebile li na le chelete e ngata ho rala le ho etsoa ho feta lisebelisoa tse tloaelehileng tsa LOCC, athe karabelo e ikemetse haholo ebile e tšoana;leha ho le joalo, ka lebaka la matla a phahameng a mochini oa lisebelisoa tsa centrifugal, lisebelisoa tse fumanehang tsa chip li fokotsehile mme meqolo e nyane e thata.ho koloi.Ka nako e ts'oanang, lisebelisoa tse ngata tsa LOAD li etselitsoe tšebeliso e le 'ngoe feela, e theko e boima bakeng sa ho lemoha ka bongata [96, 117, 118, 119].
Lilemong tse mashome tsa morao tjena, LOAD, e nkoang e le e 'ngoe ea lisebelisoa tsa microfluidic tse tšepisang ka ho fetisisa, e fumane tlhokomelo e kholo ho tsoa ho bafuputsi le bahlahisi.Ka hona, LOAD e se e amohetsoe ka bongata mme e sebelisitsoe bakeng sa tlhahlobo ea limolek'hule tsa likokoana-hloko tse tšoaetsanoang [120, 121, 122, 123, 124], haholo nakong ea koluoa ea COVID-19.Mohlala, qetellong ea 2020, Ji et al.[60] e bontšitse tlhahlobo e tobileng ea RT-qPCR bakeng sa ho lemoha ka potlako le ka mokhoa o ikemetseng oa SARS-CoV-2 le tšoaetso ea ntaramane A le B mefuteng ea 'metso.Joale Xiong et al.[74] e hlahisitse sethala sa LAMP se kopantsoeng sa discoid microfluidic bakeng sa ho lemoha ka potlako, ho nepahetseng, le ka nako e le 'ngoe ea li-coronavirus tse supileng tsa phefumoloho ea batho, ho kenyeletsoa SARS-CoV-2, nakong ea metsotso e 40.Mathoasong a 2021, de Oliveira et al.[73] e bonts'itse chip ea polystyrene toner centrifugal microfluidic, e sebelisoang ka letsoho ka rotator ea monoana, bakeng sa tlhahlobo ea molek'hule ea RT-LAMP ea COVID-19.Ka mor'a moo, Dignan et al.[39] e hlahisitse mochine o nkehang habobebe oa centrifuge microdevice bakeng sa tlhoekiso ea SARS-CoV-2 RNA ka kotloloho ho tsoa likarolong tsa buccal swab.Medved et al.[53] e sisintse tsamaiso ea sampole ea SARS-CoV-2 ea inline ea aerosol e nang le chip e nyane e potolohang ea microfluidic fluorescent e nang le moeli oa ho lemoha oa likopi tse 10/μL le moeli oa potoloho oa metsotso e 15.Suarez le ba bang.[75] haufinyane e tlalehile nts'etsopele ea sethala se kopaneng sa modular centrifugal microfluidic bakeng sa ho lemoha ka ho toba SARS-CoV-2 RNA ka lisampole tsa nasopharyngeal swab tse sa sebetsaneng le mocheso tse sebelisang LAMP.Mehlala ena e bonts'a melemo le ts'episo e kholo ea LOAD tlhahlobong ea limolek'hule tsa COVID-19.
Ka 1945 Muller le Clegg [125] ba ile ba hlahisa likanale tsa microfluidic ka lekhetlo la pele pampiring ba sebelisa pampiri ea sefa le parafine.Ka 2007, sehlopha sa Whitesides [126] se thehile sethala sa pele sa pampiri se sebetsang bakeng sa tlhahlobo ea liprotheine le tsoekere.Pampiri e fetohile substrate e loketseng bakeng sa microfluidics.Pampiri e na le thepa ea tlhaho e kang hydrophilicity le porous structure, biocompatibility e babatsehang, boima bo bobebe, ho feto-fetoha ha maemo, ho phutholoha, litšenyehelo tse tlaase, boiketlo ba tšebeliso le boiketlo.Classical µPADs e na le meaho ea hydrophilic/hydrophobic e hahiloeng holim'a likaroloana tsa pampiri.Ho itšetlehile ka sebopeho sa mahlakore a mararo, μPADs e ka aroloa ka likarolo tse peli (2D) le tse tharo-dimensional (3D) μPADs.2D µPADs e hlahisoa ka ho etsa meeli ea hydrophobic ho theha likanale tsa microfluidic, ha 3D µPADs hangata e entsoe ka mekotla ea pampiri ea 2D microfluidic, ka linako tse ling ka ho mena pampiri, mekhoa ea ho thella, likanale tse bulehileng, le khatiso ea 3D [96].Mekelikeli e nang le metsi kapa ea likokoana-hloko ho μPAD e laoloa haholo-holo ke matla a capillary ntle le mohloli oa matla a ka ntle, ho thusa ho boloka pele ho li-reagents, ho tšoara sampuli, le ho fumanoa ka makhetlo a mangata.Leha ho le joalo, taolo e nepahetseng ea phallo le tlhahlobo ea li-multiplex li sitisoa ke lebelo le sa lekaneng la ho lemoha, kutloisiso, le ts'ebeliso e ncha [96, 127, 128, 129, 130].
Joalo ka sethala se sa tloaelehang sa microfluidic, μPAD e khothalelitsoe haholo le ho ntlafatsoa bakeng sa tlhahlobo ea limolek'hule ea mafu a tšoaetsanoang a kang HCV, HIV, le SARS-CoV-2 [131, 132].Bakeng sa ho khetholla le ho lemoha ha HCV, Tengam et al.[133] e hlahisitse biosensor e ncha e thehiloeng pampiring ea fluorescent e sebelisang probe e khethehileng ea nucleic acid e thehiloeng ho pyrrolidinyl peptide.Nucleic acids ha e sebetse hantle pampiring ea selulose e nang le oxidized ka mokhoa o fokotsang alkylation lipakeng tsa lihlopha tsa amino le lihlopha tsa aldehyde, 'me ho fumanoa ho ipapisitsoe le fluorescence.Matšoao ana a ka baloa ke sesebelisoa se entsoeng ka ho khetheha se nang le k'hamera e nkehang ea fluorescent hammoho le khamera ea selefouno.Ka mor'a moo, Lu et al.[134] e thehile electrode e feto-fetohang e thehiloeng pampiring e thehiloeng holim'a nickel / khauta nanoparticles / carbon nanotubes / polyvinyl alcohol organometallic framework composite bakeng sa ho lemoha sepheo sa HIV ka ho kopanya DNA ho sebelisa methylene blue e le DNA redox indicator.Haufinyane tjena, Chowdury et al.[135] e hlahisitse moralo o inahaneloang oa sethala bakeng sa tlhahlobo ea ntlha ea tlhokomelo ea µPAD ho sebelisoa mathe a tala a mokuli hammoho le LAMP le theknoloji e nkehang ea litšoantšo bakeng sa tlhahlobo ea analyte ea COVID-19.
Liteko tsa phallo ea morao-rao li tataisa maro ka matla a capillary le ho laola motsamao oa mokelikeli ka ho koloba le litšoaneleho tsa li-porous kapa microstructured substrates.Lisebelisoa tsa lateral flow li na le sampole, conjugate, incubator le ho lemoha, le lipache tse monyang.Limolek'hule tsa nucleic acid tse LFA li lemoha li-binders tse khethehileng tse bolokiloeng pele sebakeng se tlamang 'me li tlamahane e le li-complexes.Ha mokelikeli o ntse o feta ka lipoleiti tsa incubation le ho lemoha, li-complexes li nkoa ke limolek'hule tsa ho hapa tse teng meleng ea teko le ho laola, ho bontša liphello tse ka baloang ka ho toba ka mahlo.Ka tloaelo, LFA e ka phethoa ka metsotso e 2-15, e potlakileng ho feta ho sibolloa ha setso.Ka lebaka la mochine o khethehileng, LFA e hloka ts'ebetso e fokolang 'me ha e hloke lisebelisoa tse eketsehileng, e leng se etsang hore e sebelisoe haholo.Ho bonolo ho e etsa le ho e fokotsa, 'me litšenyehelo tsa li-substrates tse entsoeng ka pampiri li tlaase.Leha ho le joalo, e sebelisoa feela bakeng sa tlhahlobo ea boleng, 'me ho lemoha bongata ho thata haholo,' me bokhoni ba multiplexing le ho feta li fokotsehile haholo, 'me ke nucleic acid e le' ngoe feela e lekaneng e ka fumanoang ka nako [96,110,127].
Leha lits'ebetso tse ngata tsa LFA li tsepamisitse maikutlo ho li-immunoassays, ts'ebeliso ea LFA bakeng sa tlhahlobo ea limolek'hule ho li-chips tsa microfluidic le eona e sebetsa ebile e tumme [136].Tabeng ea kokoana-hloko ea hepatitis B, HIV le SARS-CoV-2 LFA Gong et al.[137] e khothalelitse sethala sa nanoparticle LFA se feto-fetohang 'me se bonts'a ho feto-fetoha ha sethala sena sa miniaturized le se nkehang habonolo ka ho lemoha ka hloko le ka bongata ba lipheo tse ngata tse kang HBV nucleic acid.Ho phaella moo, Fu et al.[138] e bonts'itse buka ea LFA e thehiloeng holim'a ponelopele ea Raman e ntlafalitsoeng bakeng sa tlhahlobo ea bongata ba HIV-1 DNA maemong a tlase.Bakeng sa ho lemoha ka potlako le ka hloko ha SARS-CoV-2, Liu et al.[85] e hlahisitse tlhahlobo ea microfluidic-integrated RPA lateral flow analysis ka ho kopanya RT-RPA le mokhoa oa ho lemoha oa lateral flow ho kena tsamaisong e le 'ngoe ea microfluidic.
Ts'ebeliso ea li-platform tse fapaneng tsa microfluidic e fapana ho latela lithuto tse ikhethileng, ho nka monyetla o felletseng oa bokhoni le melemo ea sethala.E nang le li-valve, lipompo le li-ducts tse theko e tlaase, LOCC ke sethala se pharaletseng ka ho fetisisa sa mefuta-futa ea likopo le tšebelisano-'moho le sebaka se seholohali sa nts'etsopele.Ka hona, re tšepa le ho khothaletsa hore liphuputso tse ncha li etsoe LOCC e le teko ea pele le hore maemo a ntlafatsoe.Ho phaella moo, ho lebeletsoe hore mekhoa e sebetsang hantle le e nepahetseng e tla fumanoa le ho sebelisoa tsamaisong.LOAD e ipabola taolong e nepahetseng ea maro a tsoang lisebelisoa tse teng tsa LOCC mme e bonts'a melemo e ikhethang ho li-drive tse le 'ngoe ka matla a centrifugal ntle le tlhoko ea li-drive tsa kantle, athe likarabo tse bapileng li ka arohanngoa le ho hokahanngoa.Kahoo, nakong e tlang, LOAD e tla fetoha sethala se seholo sa microfluidic se nang le ts'ebetso e fokolang ea matsoho le theknoloji e hōlileng tsebong le e ikemetseng.Sethala sa µPAD se kopanya melemo ea LOCC le lisebelisoa tse thehiloeng pampiring bakeng sa litšenyehelo tse tlase, tlhahlobo ea ts'ebeliso e le 'ngoe.Ka hona, nts'etsopele ea nako e tlang e lokela ho tsepamisa maikutlo ho theknoloji e bonolo le e thehiloeng hantle.Ho feta moo, LFA e loketse hantle bakeng sa ho lemoha mahlo a hlobotseng, e tšepisa ho fokotsa tšebeliso ea sampole le ho potlakisa ho lemoha.Papiso e qaqileng ea sethala e bontšoa ho Lethathamo la 2.
Litlhahlobo tsa dijithale li arola sampole ho li-microreactors tse ngata, e 'ngoe le e' ngoe e na le palo e hlakileng ea limolek'hule tse shebiloeng [139, 140].Litlhahlobo tsa dijithale li fana ka melemo ea bohlokoa bakeng sa ho etsa palo e felletseng ka ho etsa liteko tse likete tse bapileng tsa biochemical ka nako e le 'ngoe le ka bonngoe ka likarolo tsa sekala sa micron ho fapana le mohato o tsoelang pele.Ha ho bapisoa le li-microfluidi tsa setso, karabelo ea phaposi e ka fokotsa molumo oa sampole, ea eketsa katleho ea karabelo, 'me ea kopanngoa habonolo le mekhoa e meng ea tlhahlobo ntle le tlhoko ea liteishene, lipompo, li-valve le meralo e kopaneng [141, 142, 143, 144, 145, 146, 147] .Mekhoa e 'meli e latelang e sebelisoa litlhahlobong tsa dijithale ho fihlela karohano e ts'oanang le e nepahetseng ea tharollo, ho kenyelletsa le li-reagents le disampole tse kang lisele, li-nucleic acid, le likaroloana tse ling kapa limolek'hule: (1) li-emulsions tsa lerotholi tse sebelisang ho se tsitse ha sebopeho sa mokelikeli;(2) karohano ea lihlopha e etsoa ke lithibelo tsa geometri tsa sesebelisoa.Mokhoa oa pele, marotholi a nang le li-reagents le lisampole ho li-microchannels a ka etsoa ka mekhoa e sa sebetseng e kang co-current, crossflow, flow focusing, staged emulsification, microchannel emulsification, le membranes ka matla a shear viscous le emulsification ka phetoho ea mocha.localization [143, 145, 146, 148, 149] kapa ho sebelisa mekhoa e sebetsang [150, 151], e hlahisang matla a eketsehileng ka matla a motlakase, a khoheli, a mocheso le a mechine.Mokhoeng oa ho qetela, mokhoa o motle ka ho fetisisa oa mokelikeli o lekanang ka likamoreng tsa microfluidic o arolelanoa ka ho boloka mehaho ea sebaka sa boholo bo lekanang, e kang li-micropits le li-arrays [152,153,154].Haholo-holo, marotholi ke likarolo tse kholo tsa phallo tse ka hlahisoang le ho sebelisoa ka li-electrode arrays tse thehiloeng ho digital microfluidics (DMF).Electrowetting of dielectrics ke e 'ngoe ea likhopolo-taba tsa DMF tse ithutoang ka ho fetisisa, kaha electrowetting ea dielectrics e lumella ho qhekella ka mokhoa o nepahetseng oa marotholi a motho ka mong, ho laola sebōpeho sa matšoao a motlakase a metsi le a asymmetric a fetang mahlakoreng a sa tšoaneng [141, 144].Mesebetsi e ka sehloohong e nang le marotholi a DMF e kenyelletsa ho hlophisa, ho arola le ho kopanya [151, 155, 156], e ka sebelisoang likarolong tse sa tšoaneng tsa tlhahlobo, haholo-holo ho lemoha limolek'hule [157, 158, 159].
Digital nucleic acid discovery ke tekhenoloji ea tlhahlobo ea limolek'hule ea moloko oa boraro e latelang PCR e tloaelehileng le PCR ea nako ea nnete (qPCR), e tsamaisanang le tatellano e phahameng le biopsy ea metsi.Lilemong tse mashome a mabeli tse fetileng, li-nucleic acid tsa digital li tsoetse pele ka potlako lefapheng la tlhahlobo ea limolek'hule tsa likokoana-hloko tse tšoaetsanoang [160, 161, 162].Palo e felletseng ea tlhahlobo ea dijithale ea nucleic acid e qala ka ho paka lisampole le li-reagents ka har'a likarolo tsa motho ka mong ho netefatsa hore tatellano e 'ngoe le e' ngoe ea sepheo e na le monyetla o tšoanang oa ho kena phaposing ka 'ngoe.Ho latela maikutlo, karolo ka 'ngoe e ka abeloa tatellano ea liphofu tse ngata, kapa ho kanna ha se be le sistimi e ikemetseng ea microreaction.Ka mekhoa e fapaneng ea ho lemoha e hlalositsoeng ka holimo, likarolo tse nang le tatellano ea sepheo sa likokoana-hloko tse hlahisang matšoao ka holimo ho moeli o itseng li ka bonoa ka leihlo le hlobotseng kapa ka mochine 'me li ngotsoe e le tse ntle, ha likarolo tse ling tse hlahisang matšoao ka tlase ho moeli li ngotsoe e le tse ntle. .tse mpe, tse etsang hore sesupo sa karolo ka nngwe e be boolean.Kahoo, ka ho bala palo ea likarolo tse entsoeng le sekhahla sa liphetho tse ntle ka mor'a karabelo, likopi tsa pele tsa lisampole tsa tlhahlobo li ka bapisoa ho sebelisoa foromo ea kabo ea Poisson ntle le tlhokahalo ea curve e tloaelehileng, e hlokahalang bakeng sa litlhahlobo tse tloaelehileng tsa palo e joalo. joalo ka qPCR.[163] Ha ho bapisoa le mekhoa e tloaelehileng ea ho hlahloba limolek'hule, ho lemoha ha nucleic acid ea digital ho na le tekanyo e phahameng ea ho iketsetsa, lebelo le phahameng la tlhahlobo le kutloisiso, li-reagents tse fokolang, tšilafalo e fokolang, le moralo o bonolo le ho etsoa.Ka mabaka ana, ts'ebeliso ea litlhahlobo tsa dijithale, haholo mekhoa e theoleloang, bakeng sa tlhahlobo ea limolek'hule, ho kopanya mekhoa ea ho holisa le ho bala matšoao, e ithutoe hantle nakong ea ho phatloha ho hoholo ha SARS-CoV-2.Ka mohlala, Yin et al.[164] mekhoa e kopantsoeng ea li-droplet tsa digital le e potlakileng ea PCR ho lemoha mefuta ea ORF1ab, N, le RNase P ho SARS-CoV-2 ka har'a chip ea microfluidic.Ka ho hlakileng, tsamaiso e ile ea khona ho khetholla letšoao le letle ka har'a metsotsoana ea 115, e potlakileng ho feta PCR e tloaelehileng, e bontšang katleho ea eona ho lemoha ntlha ea tlhokomelo (Setšoantšo sa 7a).Dong et al.[165], Jala et al.[157], Chen et al.[166] le Alteri et al.[167] e boetse e sebelisa Droplet digital PCR (ddPCR) ho bona SARS-CoV-2 tsamaisong ea microfluidic e nang le liphello tse tsotehang.Ho tsoela pele ho ntlafatsa sekhahla sa ho lemoha, Shen et al.[168] e fihletse ddPCR-based chip imaging ka nako e nyane joalo ka 15 s ntle le tšebeliso ea mekhoa ea ho roka litšoantšo, ho potlakisa ts'ebetso ea theknoloji ea ddPCR ho tloha lab ho ea ho kopo.Ha se feela mekhoa ea ho holisa mocheso joalo ka PCR, empa hape le mekhoa ea amplification ea isothermal e sebelisoa ho nolofatsa maemo a karabelo le karabelo e potlakileng.Lu et al.[71] e hlahisitse SlipChip bakeng sa tlhahlobo ea marotholi, e khonang ho hlahisa marotholi a boholo bo fapaneng ka sekhahla se phahameng ka mohato o le mong le ho lekanya SARS-CoV-2 nucleic acids ho sebelisa LAMP ea digital (Setšoantšo sa 7b).Joalo ka mahlale a ntseng a fetoha ka potlako, CRISPR le eona e ka bapala karolo ea bohlokoa ho sibolloeng ha nucleic acid ea dijithale ka litšoantšo tse bonolo tsa colorimetric ntle le tlhoko ea matheba a eketsehileng a nucleic acid.Ackerman le ba bang.e hlahisitse karabelo ea matrix e kopaneng bakeng sa tlhahlobo ea multiplex ea li-nucleic acid.[158] e lemohile livaerase tse 169 tse amanang le batho, ho kenyeletsoa SARS-CoV-2, marotholing a nang le li-reagents tsa CRISPR-Cas13 tse thehiloeng ho nucleic acid ka tlhahlobo ea microwell (Setšoantšo sa 7c).Ho phaella moo, isothermal amplification le theknoloji ea CRISPR e ka sebelisoa tsamaisong e le 'ngoe ho kopanya melemo ea bobeli.Park et al.[169] Tekolo ea dijithale ea CRISPR/Cas12a e entsoe ka chip ea khoebo ea microfluidic bakeng sa ho sibolla SARS-CoV-2 e ntšitsoeng le e bolailoeng ke mocheso e ipapisitse le RT-RPA ea mothati o le mong e nang le tšibollo e khuts'oane le e phahameng ea lets'oao ho ea morao. karo-karolelano ea nako., mefuta e mengata e matla le kutloisiso e molemo (setšoantšo sa 7d).Litlhaloso tse ling tsa mehlala ena li fanoe ho Lethathamo la 3.
Sethala se tloaelehileng sa dijithale bakeng sa ho lemoha ha nucleic acid.a Ts'ebetso e potlakileng ea PCR ea dijithale e na le mehato e mene ea bohlokoa: tokiso ea sampole, kabo ea motsoako oa karabelo, ts'ebetso ea ho holisa, le palo ea sepheo (e fetotsoeng ho tsoa ho [164]).b Sekema se bonts'a tlhahlobo ea marotholi a SlipChip bakeng sa sebopeho sa marotholi ka bongata bo boholo (e nkiloe ho [71]).c CARMEN-Cas sets'oants'o sa ts'ebetso ea mosebetsi13 (e nkiloe ho [158]).d Kakaretso ea tlhahlobo e tsoetseng pele ea vaerase ea dijithale ka CRISPR/Cas ka pitseng e le 'ngoe (e nkiloe ho [169]).W/O metsi-ka-oli, polydimethylsiloxane PDMS, PCR polymerase chain reaction, DAQ data collection, PID proportional integral derivative, CARMEN combinatorial matrix reaction bakeng sa tlhahlobo ea multiplex nucleic acid, SARS-CoV-2, lefu le matla le matla la ho hema, coronavirus 2 , RT Amplification ea reverse transcriptase recombinase polymerase-RPA, letšoao la S/B ka morao
Nako ea poso: Sep-15-2022
