Njira zamakono zodziwira matenda opatsirana zimafuna kugwiritsa ntchito zida za benchtop zomwe sizili zoyenera kuyesedwa kwapadera (POCT).Emerging microfluidics ndiukadaulo wocheperako kwambiri, wodzipangira okha, komanso wophatikizika womwe ndi njira ina yosinthira njira zakale zowunikira mwachangu, zotsika mtengo, zolondola pamasamba.Njira zodziwira mamolekyulu zimagwiritsidwa ntchito kwambiri pazida za microfluidic monga njira zothandiza kwambiri zodziwira tizilombo toyambitsa matenda.Ndemangayi ikufotokoza mwachidule kupita patsogolo kwaposachedwa kwa matenda a microfluidic-based molecular of matenda opatsirana kuchokera kumaphunziro ndi mafakitale.Choyamba, timafotokoza momwe ma nucleic acid amapangidwira pa-chip, kuphatikiza kusanja kwachitsanzo, kukulitsa, ndi kuwerenga kwazizindikiro.Makhalidwe, ubwino ndi kuipa kwa mitundu inayi ya mapulatifomu a microfluidic amafananizidwa.Kenako, tikambirana za kugwiritsa ntchito kuyesa kwa digito pakuwerengera kwathunthu kwa ma nucleic acid.Zida zonse zakale komanso zaposachedwa zamalonda za microfluidic-based molecule zowunikira zimafotokozedwa mwachidule ngati umboni wa momwe msika uliri.Pomaliza, tikupangira njira zamtsogolo za matenda a microfluidic a matenda opatsirana.
Matenda opatsirana amayamba ndi tizilombo toyambitsa matenda, kuphatikizapo mabakiteriya, mavairasi, ndi tizilombo toyambitsa matenda, zomwe zimafalitsidwa padziko lonse lapansi.Mosiyana ndi matenda ena, tizilombo toyambitsa matenda timayamba kutenga kachilomboka ndikufalikira pakati pa anthu ndi nyama zomwe zimagwidwa kudzera mu inoculation, mpweya ndi madzi [1].Kupewa matenda opatsirana ndikofunikira ngati njira yaumoyo wa anthu.Njira zazikulu zitatu zothanirana ndi matenda opatsirana: (1) kuwongolera magwero a matenda;(2) kusokoneza njira yopatsirana;(3) kutetezedwa kwa anthu omwe ali pachiwopsezo.Pakati pa njira zazikuluzikulu, kuwongolera gwero la matenda kumaonedwa ngati njira yofunika kwambiri chifukwa cha kuphweka kwake komanso mtengo wake wotsika.Kuzindikira mwachangu, kudzipatula, komanso kuchiza anthu omwe ali ndi kachilomboka ndikofunikira, kumafuna njira zowunikira mwachangu, zachidziwitso, komanso zolondola [2].Kuzindikirika kwa matenda opatsirana masiku ano nthawi zambiri kumaphatikiza kuyezetsa kwachipatala potengera zizindikiro ndi zizindikiro ndi maphunziro a labotale monga chikhalidwe cha ma cell ndi matenda a ma cell, omwe amafunikira anthu ophunzitsidwa bwino, njira zolimbikitsira ntchito, ndi zida zoyesera zodula [3, 4].Kupewa kufalikira kwa matenda opatsirana kumafuna matenda ofulumira, otsika mtengo, komanso olondola a m'deralo, makamaka m'madera omwe alibe zida zomwe matenda opatsirana ndi ofala komanso ovuta kwambiri [5], komanso chithandizo chamankhwala m'chipululu kapena pankhondo, kumene zochitika zadzidzidzi sizidziwikiratu..chithandizo chamankhwala chili ndi malire [6].M'nkhaniyi, ma microfluidics ndi teknoloji yomwe imaphatikizapo matekinoloje a microelectromechanical systems, nanotechnology, kapena sayansi ya zipangizo kuti azitha kuyendetsa bwino madzimadzi [7,8,9,10], kupereka mwayi watsopano wodziwikiratu (POCT).) mankhwala opatsirana kunja kwa zipatala ndi ma laboratories.Poyerekeza ndi zowunikira zakale zomwe zimawononga nthawi, ukadaulo wa microfluidic umapereka zitsanzo komanso kupulumutsa ndalama pakuwunika kwa maselo panthawi ya miliri.Kufalikira kwapadziko lonse lapansi kwa matenda a coronavirus 2019 (COVID-19) kumayamba chifukwa chachikulu cha kupuma kwapadziko lonse lapansi kwa coronavirus 2 (SARS-CoV-2), kotero kufunikira kwa ma microfluidics pakupewa komanso kuwongolera mliriwu kumatsindikanso [11, 12]. , 13].Mosiyana ndi matenda achikhalidwe, POCT ya microfluidic imagwiritsa ntchito zida zazing'ono zosunthika kuyambira zowunikira za benchtop kupita ku timizere tating'ono tating'ono toyesa pafupi ndi sampuli [14].Mayesowa amakhala osavuta kapena osakonzekera zachitsanzo, kukulitsa ma siginecha mwachangu, komanso kuwerengeka kwazizindikiro zomwe zimapangitsa kuti zizikhala zazifupi komanso zotsatira zolondola mkati mwa mphindi.Kupezeka ndi kupanga kwakukulu kwa zida zothandizira zaumoyo zokhala ndi microfluidic zakulitsa ntchito zawo zotsika mtengo komanso zachindunji kunja kwa chipatala, pafupi ndi wodwalayo, komanso kunyumba.
Pakati pa njira zomwe zilipo zodziwira matenda opatsirana, kufufuza kwa maselo ndi chimodzi mwazovuta kwambiri [15, 16].Kuphatikiza apo, kuwunika kwa maselo nthawi zambiri kumagwiritsidwa ntchito ngati mulingo wagolide wowunikira mosalekeza COVID-19, kulola kuzindikirika mwachindunji kwa zigawo za RNA kapena DNA chitetezo cha mthupi chisanayambike [17, 18].Pakuwunika kwapano, tikuwonetsa kupita patsogolo kwaposachedwa kwambiri mu njira zowunikira ma cell a microfluidics a matenda opatsirana, kuchokera kumaphunziro apamwamba kupita kuzinthu zamtsogolo zamakampani (mkuyu 1).Tiyamba ndi masitepe atatu ofunikira pakuzindikira kwa nucleic acid: pa-chip sample pretreatment, nucleic acid amplification, ndi kuwerenga ma siginecha.Kenako tidafanizira mitundu yosiyanasiyana ya nsanja za microfluidic ndi kapangidwe kake ndi ntchito, kuwonetsa mawonekedwe apadera (mphamvu ndi zofooka).Kuzindikira kwa Digital nucleic acid kumakambidwanso ndikuperekedwa ngati chitsanzo chaukadaulo wa m'badwo wachitatu pakuwerengera kwathunthu kwa mamolekyu opatsirana.Kuphatikiza apo, zida zingapo zaposachedwa komanso zaposachedwa za POCT zidzawonetsedwa kuti ziwonetse momwe msika wa Microfluidic POCT ulili wowunikira ma cell.Tidzakambirananso ndikufotokozera masomphenya athu a ntchito zamtsogolo.
Ma module a tchipisi ta microfluidic ozindikira nucleic acid amatha kugawidwa m'magulu atatu (sampling, kuzindikira, ndi kusaina) malinga ndi ntchito zawo [19].Pakati pa ma module awa, gawo lachitsanzo makamaka limazindikira zitsanzo za lysis ndi nucleic acid m'zigawo.Sensa module makamaka amawongolera kutembenuka ndi kukulitsa ma nucleic acid ma sign.Module yowonetsera imazindikira chizindikiro chosinthidwa ndikusinthidwa ndi gawo lozindikira.Kutengera njira yodziwira ma nucleic acid pa chip, tifotokozera mwachidule ma chips osiyanasiyana omwe amatha kuzindikira ntchito ya "zolowetsa ndi kutulutsa".
Gawo loyamba pakuzindikira kwa ma nucleic acid ndikutulutsa kwa nucleic acid, mwachitsanzo, kupatula chandamale cha nucleic acid kuchokera ku zitsanzo zoyambirira.Kutulutsa kwa nucleic acid kumachitidwa kuti ayeretse ma nucleic acid kuchokera kuzinthu zina zama cell, kuonetsetsa kukhulupirika kwa kapangidwe kake ka nucleic acid mamolekyulu, ndikuwonjezera zotsatira.Kutulutsa kwa nucleic acid kumafuna chitsanzo chofunikira cha lysis ndi nucleic acid kugwidwa, khalidwe ndi mphamvu zake zomwe zimakhudza kwambiri kafukufuku ndi zotsatira za matenda.Zotsatira zilizonse zobisika panthawi yochotsa zimatha kuchepetsa kuzindikira.Mwachitsanzo, njira za polymerase chain reaction (PCR) ndi njira za loop isothermal amplification (LAMP) zimaletsedwa ndi zosungunulira zina zotsalira monga ethanol ndi isopropanol mu nucleic acid isolation reagents [20].Kutulutsa kwamadzimadzi ndi kutulutsa kolimba ndi njira zodziwika kwambiri zolekanitsira ma nucleic acids [21], komabe, kutulutsa kwamadzimadzi pa chip kumakhala kochepa kwambiri, chifukwa ma reagents omwe amagwiritsidwa ntchito pochotsa madzi amadzimadzi amachititsa dzimbiri tchipisi tating'onoting'ono tating'onoting'ono. .Apa, tikuwunikira njira zopangira gawo lolimba la ma microarray ndikuyerekeza zabwino ndi zovuta zawo.
Silicon ndi gawo laling'ono lomwe limagwirizana ndi nucleic acid chifukwa cha biocompatibility, kukhazikika, komanso kusintha kosavuta [22].Chofunika kwambiri, chikasinthidwa ndi silika kapena zipangizo zina, kaphatikizidwe kameneka kamasonyeza katundu kuti adsorbe ma nucleic acids otsika pansi pa pH yotsika, mikhalidwe yamchere yambiri pamene ili ndi pH yapamwamba, mchere wochepa.Kutengera chodabwitsa ichi, ndizotheka kuyeretsa nucleic acid.
Mitundu yosiyanasiyana ya zida za silika zakhala zikugwiritsidwa ntchito pochotsa nucleic acid mu microfluidics, monga mikanda ya silika, ufa, zosefera za microfiber, ndi nembanemba za silika [23, 24, 25, 26].Kutengera momwe zinthu ziliri, zida zopangira silicon zitha kugwiritsidwa ntchito mu ma microcircuits m'njira zosiyanasiyana.Mwachitsanzo, ma silika granules, ufa, ndi nanofilters zamalonda zitha kuyikidwa mu pores kapena ma microchannel a tchipisi tating'onoting'ono ndikuthandizira kuchotsa ma nucleic acid kuchokera ku zitsanzo [27, 28, 29].Zingwe za silika zosinthidwa pamwamba zitha kugwiritsidwanso ntchito kuyeretsa mwachangu DNA ku tizilombo toyambitsa matenda pamtengo wotsika.Mwachitsanzo, Wang et al.[30] Mwa kuphatikiza machitidwe a denaturing amplification ndi vesicle-mediated chain exchanges ndi silika nembanemba yokutidwa ndi chitosan oligosaccharides, makina osunthika osunthika adayambitsidwa omwe adazindikira bwino mayunitsi a 102-108 kupanga magulu.(CFU)/ml Vibrio parahaemolyticus., ndipo kukhalapo kwa kachilomboka kunawonekera mosavuta.Powell et al.[31] Ma microarrays opangidwa ndi silicon adagwiritsidwa ntchito kuti azindikire kachilombo ka hepatitis C (HCV), kachilombo ka HIV (HIV), kachilombo ka Zika, ndi papillomavirus yaumunthu ndi kufalitsa kokha, momwe 1.3 μl tortuous microreactor inapangidwa kuti igwire ma virus a RNA.ndikuchita in situ amplification.Kuphatikiza pa njirazi, ma microcolumns osinthidwa pamwamba a silika amathandizanso kwambiri pakuchotsa kwa nucleic acid, monga momwe geometry ndi zinthu zosinthira zimakulitsa kwambiri kutulutsa bwino.Chen et al.[32] anakonza nsanja ya microfluidic yodzipatula ya RNA yotsika kwambiri yotengera ma amino-coated silicon microcolumns.Chipangizo cha microfluidic ichi chimaphatikiza ma micropillars a 0.25 cm2 pa gawo lapansi la silicon kuti akwaniritse bwino kwambiri kutulutsa kudzera m'malo okwera mpaka ma volume.Ubwino wa kapangidwe kameneka ndikuti chipangizo cha microfluidic chimatha kufikira 95% nucleic acid m'zigawo.Njira zopangira silicon izi zikuwonetsa kufunika kodzipatula mwachangu ma nucleic acid pamtengo wotsika.Kuphatikiza ndi tchipisi tating'onoting'ono tating'onoting'ono, njira zochotsera silicon sizimangowonjezera luso la kuzindikira kwa nucleic acid, komanso zimathandizira kuti miniaturization ndi kuphatikiza zida zowunikira [20].
Njira zolekanitsa maginito zimagwiritsa ntchito maginito particles kuti alekanitse nucleic acid pamaso pa kunja kwa maginito.Zomwe zimagwiritsidwa ntchito kwambiri ndi maginito particles monga Fe3O4 kapena γ-Fe2O3 maginito particles yokutidwa ndi silika, amino ndi carboxyl [33,34,35,36].Kusiyanitsa kwa maginito maginito poyerekeza ndi njira za silicon-based SPE ndizosavuta kugwiritsa ntchito ndikuwongolera ndi maginito akunja.
Pogwiritsa ntchito kuyanjana kwa electrostatic pakati pa nucleic acid ndi silika, pansi pa mchere wambiri ndi pH yochepa, ma nucleic acids amapangidwa pamwamba pa maginito opangidwa ndi silika, pamene mchere wochepa ndi pH wambiri, mamolekyu amatha kutsukidwa. kachiwiri..Mikanda ya maginito yokhala ndi silika imapangitsa kuti zitheke kuchotsa DNA kuchokera ku zitsanzo zazikulu (400 μL) pogwiritsa ntchito kayendedwe koyendetsedwa ndi maginito [37].Monga chiwonetsero, Rodriguez-Mateos et al.[38] amagwiritsa ntchito maginito osinthika kuti aziwongolera kusamutsa kwa mikanda ya maginito kupita ku zipinda zosiyanasiyana.Kutengera ndi maginito okhala ndi silika, makope 470/mL a SARS-CoV-2 genomic RNA atha kuchotsedwa ku zitsanzo zamadzi oyipa kuti azindikire za LAMP reverse transcript (RT-LAMP) ndipo yankho limatha kuwerengedwa pakadutsa ola limodzi.diso lamaliseche (mkuyu 2a).
Zipangizo zochokera maginito ndi porous zipangizo.Chithunzi chojambula cha chipangizo cha IFAST RT-LAMP microfluidic cha SARS-CoV-2 RNA kuzindikira (chosinthidwa kuchokera ku [38]).b Centrifugal yaying'ono chipangizo cha dSPE cha buccal swab nucleic acid (yosinthidwa kuchokera ku [39]).c Makina odzipangira okha opangira mphamvu pogwiritsa ntchito FTA® khadi (yosinthidwa kuchokera ku [50]).d Fusion 5 fyuluta pepala losinthidwa ndi chitosan (losinthidwa kuchokera ku [51]).SARS-CoV-2 kwambiri pachimake kupuma matenda coronavirus 2, RT-LAMP reverse transcription loop mediated isothermal amplification, FTA finders teknoloji ogwirizana, NA nucleic acid
Tinthu tating'onoting'ono tokhala ndi maginito ndi abwino kumangirira msana wa phosphate wa nucleic acid.Pa ndende ina yamchere, magulu a phosphate omwe ali ndi vuto la nucleic acid amatha kuyimbidwa bwino pamtunda wa tinthu tating'ono ta maginito.Choncho, maginito nanoparticles ndi akhakula pamwamba ndi mkulu kachulukidwe amino magulu anapangidwa kuti m'zigawo nucleic zidulo.Pambuyo pa kupatukana kwa maginito ndi kutsekereza, maginito nanoparticles ndi ma DNA complexes atha kugwiritsidwa ntchito mwachindunji mu PCR, zomwe zimathetsa kufunikira kwa ntchito zovuta komanso zowononga nthawi komanso kuyeretsa [35].Maginito nanoparticles okutidwa ndi zoipa carboxyl magulu agwiritsidwanso ntchito kulekanitsa nucleic zidulo adsorbed pa pamwamba ndende polyethylene glycol ndi sodium kolorayidi njira [36].Ndi mikanda ya maginito yosinthidwa pamwambayi, kutulutsa kwa DNA kumagwirizana ndi kukulitsa kotsatira.Dignan et al.[39] adalongosola pulatifomu yokhazikika komanso yosunthika ya centrifugal microfluidic ya nucleic acid pretreatment, kulola osagwiritsa ntchito luso kuti agwiritse ntchito patsamba.Kuonjezera apo, kuyanjana kwa DNA yokhayokha ndi LAMP, njira yoyenera yowunikira mfundo ya nucleic acid, ikuwonetseranso zofunikira zochepa za zipangizo ndi kuyenerera kwa mitundu ya colorimetric (mkuyu 2b).
Njira zamaginito za mikanda zimapereka mwayi wodzipangira zokha, zina zomwe zimakhalapo muzamalonda zama nucleic acid extractors [KingFisher;ThermoFisher (Waltham, MA, USA), QIAcube® HT;CapitalBio (Beijing, China) ndi Biomek®;Beckman (Miami, USA).), Florida, USA)].Ubwino wophatikiza mikanda ya maginito ndi ma microfluidics angagwiritsidwe ntchito pochotsa ma nucleic acids, omwe atha kupititsa patsogolo kuwunika kwa maselo;komabe, kuphatikiza kwa mikanda ya maginito yokhala ndi ma microfluidics kumadalirabe kwambiri machitidwe owongolera ovuta kuti azitha kuwongolera bwino mikanda ya maginito, zomwe zimafotokoza kutchuka kwa zinthu zamalonda kukhala zochulukira komanso zodula, zomwe zimalepheretsa kugwiritsa ntchito mikanda ya maginito ku POCT.
Zida zingapo za porous monga zosefera za nitrocellulose zosinthidwa, makhadi a Finders Technology Associates (FTA), mapepala osefera opangidwa ndi polyethersulfone, ndi zida zopaka glycan zagwiritsidwanso ntchito pozindikira nucleic acid [40, 41, 42, 43, 44].Zinthu za porous fibrous monga mapepala a ulusi zinayamba kugwiritsidwa ntchito polekanitsa DNA pomangirira mamolekyu aatali aatali a DNA ndi ulusi.Ma pores ang'onoang'ono amatsogolera ku kuletsa kwamphamvu kwa mamolekyu a DNA, omwe amakhudza kutulutsa kwa DNA.Chifukwa cha kukula kosiyanasiyana kwa mapepala a ulusi, kutulutsa bwino sikungakwaniritse zosowa za DNA amplification [45, 46].Khadi la FTA ndi pepala losefera zamalonda lomwe limagwiritsidwa ntchito pazamankhwala azachipatala ndipo limagwiritsidwa ntchito kwambiri m'malo ena owunikira ma cell.Pogwiritsa ntchito pepala la cellulose fyuluta yomwe imayikidwa ndi mankhwala osiyanasiyana kuti lyse ma cell a cell mu chitsanzo, DNA yotulutsidwa imatetezedwa ku kuwonongeka kwa zaka ziwiri.Posachedwapa, pepala lopangidwa ndi cellulose lapangidwa kuti lizindikire tizilombo toyambitsa matenda osiyanasiyana, kuphatikizapo SARS-CoV-2, leishmaniasis, ndi malungo [47,48,49].Kachilombo ka HIV mu plasma yokhayokha ndi lysed mwachindunji, ndipo viral nucleic acid imapindula mu FTA® yothamanga nembanemba yomwe imapangidwira mu concentrator, yomwe imalola kupanga bwino kwa nucleic acid [50] (Mkuyu 2c).Vuto lalikulu pakuzindikira kwa ma nucleic acid pogwiritsa ntchito makhadi a FTA ndikuti mankhwala monga guanidine ndi isopropanol amalepheretsa kukulitsa kotsatira.Kuti tithane ndi vutoli, tidapanga pepala la Fusion 5 la chitosan-modified filter, lomwe limaphatikiza ubwino wa kuphatikizika kwa ma molekyulu a DNA ndi pepala losefera ulusi, komanso kutulutsa kwa DNA pamagetsi osinthidwa a chitosan kuti tikwaniritse kutulutsa kwa nucleic acid. ..zosefera ulusi [51] (mkuyu 2d).Mofananamo, Zhu et al.[52] adawonetsa njira ya PCR yosinthidwa ndi chitosan yotengera in situ capillary microfluidic system yodzipatula mwachangu ndikuzindikira Zika virus RNA.Ma nucleic acids amatha kukhala adsorbed / desorbed mu lysate / PCR sing'anga, motero, kutengera / kuzimitsa lophimba katundu wa chitosan.on and off”, kuyankha pH.
Monga tafotokozera pamwambapa, njirazi zimaphatikiza ubwino wa zida zosiyanasiyana zolimba ndikuwonjezera mphamvu ya nucleic acid m'zigawo mu microfluidics.Muzochita zogwiritsidwa ntchito, kugwiritsa ntchito zipangizozi mochuluka ndizopanda ndalama, ndipo chithandizo choyenera chapamwamba kapena kusintha kwapamwamba kwa zinthu zomwe zimagwiritsidwa ntchito ndi zipangizozi zingathenso kusunga ntchito yawo.Choncho, akukhulupirira kuti kukhazikitsidwa kwa njirazi pambuyo pa kafukufuku woyendetsa ndege kungachepetse ndalama.
Kuyesa kwa ma nucleic acid pamapulatifomu a microfluidic nthawi zambiri kumagwiritsa ntchito ma voliyumu ang'onoang'ono (<100 µl), chifukwa chake kumafuna kukulitsa chandamale cha ma nucleic acid omwe ali ndi ma probes apadera kuti asinthe kukhala chizindikiro chomwe chili chosavuta kuzindikira kumunsi kwa mtsinje (mawonekedwe, magetsi, ndi maginito) [53, 54]. Kuyesa kwa ma nucleic acid pamapulatifomu a microfluidic nthawi zambiri kumagwiritsa ntchito ma voliyumu ang'onoang'ono (<100 µl), chifukwa chake kumafuna kukulitsa chandamale cha ma nucleic acid omwe ali ndi ma probes apadera kuti asinthe kukhala chizindikiro chomwe chili chosavuta kuzindikira kumunsi kwa mtsinje (mawonekedwe, magetsi, ndi maginito) [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]. Poyesa ma nucleic acids pamapulatifomu a microfluidic, ma voliyumu ang'onoang'ono (<100 µL) amagwiritsidwa ntchito nthawi zambiri, kotero kukulitsa kwa chandamale cha nucleic acid ndi ma probe apadera kumafunika kuti asinthe kukhala chizindikiro chosavuta kuti chizindikirike (zowoneka, zamagetsi, ndi maginito) [53, 54].微流控微流控 phhrup P 的 线上].微流控微流控 ph P 的 线上 的 使用 ((<μ), 因此]. 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]]. Kuzindikira ma nucleic acid pamapulatifomu a microfluidic nthawi zambiri kumagwiritsa ntchito zitsanzo zazing'ono (<100 μl), zomwe zimafunika kukulitsa chandamale cha ma nucleic acid okhala ndi ma probe apadera kuti awasinthe kukhala ma siginecha kuti adziwike (zowoneka, zamagetsi, ndi maginito) [53, 54]] .Nucleic acid amplification mu microfluidics imathanso kufulumizitsa machitidwe, kukhathamiritsa malire ozindikira, kuchepetsa zofunikira zachitsanzo, ndikuwongolera kuzindikira kolondola [55, 56].M'zaka zaposachedwa, pozindikira kuzindikira mwachangu komanso molondola, njira zingapo zokulitsa ma nucleic acid zakhala zikugwiritsidwa ntchito mu microfluidics, kuphatikiza PCR ndi machitidwe ena a isothermal amplification.Gawoli lifotokoza mwachidule njira zodziwira ma nucleic acid potengera ma microfluidic system.
PCR ndi kayeseleledwe ka DNA replication ndondomeko ya chamoyo, chiphunzitso chake chafotokozedwa mwatsatanetsatane kwina ndipo sitidzakambidwa pano.PCR imatha kukulitsa kachulukidwe kakang'ono ka chandamale cha DNA/RNA pamlingo wokulirapo, kupangitsa PCR kukhala chida champhamvu chozindikira mwachangu ma nucleic acid.M'zaka makumi angapo zapitazi, zida zambiri zonyamulika za microfluidic zokhala ndi makina oyendetsa njinga zamoto a PCR zapangidwa kuti zikwaniritse zofunikira pakuwunika kosamalira [57, 58].Pa-chip PCR imatha kugawidwa m'mitundu inayi (yokhazikika, yoyenda mosalekeza, yosinthika pang'ono, ndi PCR yolumikizira) molingana ndi njira zowongolera kutentha [59].Mwachitsanzo, Gee et al.[60] adapanga njira yachindunji yosinthira kuchulukira kwa PCR (RT-qPCR) pa nsanja yawo ya microfluidic yodziwira kuchuluka kwa SARS-CoV-2, ma virus a fuluwenza A ndi B mu zitsanzo za swab zapakhosi (Mkuyu 3a) .Park et al.[61] anamanga chipangizo chosavuta chowunikira tizilombo pophatikiza filimu yopyapyala ya PCR, ma electrode, ndi polydimethylsiloxane-based microfluidic module yopangidwa ndi chala.Komabe, ntchito zonsezi zimakhala ndi zolakwika zomwe zimachitika pa PCR wamba.PCR imafuna kupalasa njinga zotentha, zomwe zimalepheretsa kachipangizo kakang'ono kachipangizo komanso kuchepetsa nthawi yoyesera.
Kupanga kopitilira muyeso kochokera ku microfluidic ndi PCR yosinthira malo ndikofunikira kuti tithane ndi vutoli.Pogwiritsa ntchito njira yayitali ya serpentine kapena kanjira kakang'ono kowongoka, PCR yosalekeza imatha kupereka kukulitsa mwachangu pozungulira mozungulira ma reagents m'magawo atatu a preheat okhala ndi pampu yopanda chip.Opaleshoniyi imapewa bwino gawo la kusintha pakati pa kutentha kosiyanasiyana ndipo motero kumachepetsa kwambiri nthawi yoyesera [62] (Mkuyu 3b).Mu kafukufuku wina wa Jung et al.[63] anakonza njira yatsopano yosinthira ma genetic ya PCR yomwe imaphatikiza mawonekedwe a PCR osasunthika komanso otaya kwa ultrafast ndi multiplex reverse transcription PCR (mkuyu 3c).Kwa nucleic acid amplification, PCR microchip idzazunguliridwa kupyolera muzitsulo zitatu zotentha pa kutentha kosiyana: 1. Denaturation block 94 ° C, 2. Annealing block pa 58 ° C, 3. Chowonjezera chowonjezera pa 72 ° C.
Kugwiritsa ntchito PCR mu microfluidics.Kuyimilira kwadongosolo kwa dirRT-qPCR papulatifomu ya microfluidic (yosinthidwa kuchokera ku [60]).b Kuyimira kwadongosolo kwa PCR microarray yosalekeza yotengera njira ya serpentine (yosinthidwa kuchokera ku [62]).c Kuyimira kwadongosolo kwa rotary PCR genetic analyzer, yopangidwa ndi microchip, zotchingira zitatu zotenthetsera ndi stepper motor (yosinthidwa kuchokera ku [63]).d Chithunzi cha thermoconvection PCR yokhala ndi centrifugation ndi khwekhwe (yosinthidwa kuchokera [64]).DirRT-qPCR, molunjika kuchuluka kwa reverse transcript polymerase chain reaction
Pogwiritsa ntchito ma capillaries ndi malupu kapenanso mbale zoonda, convection PCR imatha kukulitsa ma nucleic acids ndi matenthedwe achilengedwe aulere popanda kufunikira kwa mpope wakunja.Mwachitsanzo, cyclic olefin polymer microfluidic platform inapangidwa pa siteji yotentha yozungulira yomwe imagwiritsa ntchito njinga yamoto ndi centrifugation mu PCR loop microchannel [64] (Mkuyu 3d).Njira yothetsera vutoli imayendetsedwa ndi kutentha kwa kutentha, komwe kumasinthasintha kutentha ndi kutentha pang'ono mu microchannel yokhala ndi annular.Ntchito yonse yokulitsa imatha kumalizidwa mu mphindi 10 ndi malire a 70.5 pg/channel.
Monga zikuyembekezeredwa, PCR yofulumira ndi chida champhamvu chophatikizira bwino zitsanzo-mayankhidwe a maselo owunikira ndi ma multiplex.Rapid PCR imachepetsa kwambiri nthawi yofunikira kuti izindikire SARS-CoV-2, zomwe zimathandizira kuwongolera bwino mliri wa COVID-19.
PCR imafuna chozungulira chotenthetsera chomwe sichiyenera POCT.Posachedwapa, njira za isothermal amplification zakhala zikugwiritsidwa ntchito ku microfluidics, kuphatikizapo koma osati ku LAMP, recombinase polymerase amplification (RPA), ndi kukulitsa kutengera nucleic acid sequences [65,66,67,68].Ndi njira izi, ma nucleic acid amachulukitsidwa ndi kutentha kosalekeza, zomwe zimathandizira kupanga zida zotsika mtengo, zodziwika kwambiri za POCT zowunikira ma cell.
Mayeso apamwamba a LAMP opangidwa ndi microfluidics amalola kuti azindikire matenda opatsirana ambiri [42, 69, 70, 71].Kuphatikizana ndi centrifugal microfluidic system, LAMP imatha kupititsa patsogolo kuzindikira kwa nucleic acid [69, 72, 73, 74, 75].SlipChip yozungulira-ndi-react idapangidwa kuti iwonetsere mabakiteriya angapo ofanana pogwiritsa ntchito LAMP [76] (Mkuyu 4a).Mukamagwiritsa ntchito LAMP yokhathamiritsa pakuyesa, chiŵerengero cha fluorescence-to-phokoso chinali pafupifupi 5, ndipo malire ozindikira adafika makope 7.2 / μl a genomic DNA. Komanso, kukhalapo kwa tizilombo toyambitsa matenda tomwe timayambitsa kugaya chakudya, kuphatikiza Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis ndi Vibrio parahaemolyticus, adawonetsedwa potengera njira mu <60 min. Komanso, kukhalapo kwa tizilombo toyambitsa matenda tomwe timayambitsa kugaya chakudya, kuphatikiza Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis ndi Vibrio parahaemolyticus, adawonetsedwa potengera njira mu <60 min.Komanso, kupezeka kwa mabakiteriya asanu omwe amapezeka m'matumbo am'mimba, kuphatikiza Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis ndi Vibrio parahaemolyticus, adawonetsedwa pogwiritsa ntchito njirayi pasanathe mphindi 60.此外, 基于 该 该 在 在 在 在 分钟 分钟 内 视化 了 种 常见 的 原体 原体 肠杆菌蜡状此外, 基于 该 该 在 在 在 在 分钟 分钟 视化 了 了 的 细菌病 细菌病 肠杆菌 肠杆菌 肠杆菌 芽孢杆菌 芽孢杆菌 肠杆菌 肠杆菌 肠杆菌 肠杆菌 肠杆菌 肠杆菌 肠杆菌 肠杆菌 和 和弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 HIPKuonjezera apo, kukhalapo kwa tizilombo toyambitsa matenda a m'mimba asanu, kuphatikizapo Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvius, ndi Vibrio parahaemolyticus, adawonetsedwa pogwiritsa ntchito njirayi pasanathe mphindi 60.
Ubwino wa LAMP mu microfluidics umaphatikizapo, pakati pa ena, kuyankha mwachangu komanso kuzindikira kocheperako.Komabe, chifukwa cha kutentha komweko (kuzungulira 70 ° C), ma aerosols amapangidwa nthawi ya LAMP, zomwe zimapangitsa kuti pakhale chiwopsezo chabodza.Mawonekedwe a Assay, mapangidwe oyambira, komanso kuwongolera kutentha kumafunikanso kukonzedwanso pa LAMP.Kuphatikiza apo, mapangidwe a chip omwe amagwiritsa ntchito kuzindikira chandamale kangapo pa chip imodzi ndiofunika kwambiri ndipo akuyenera kupangidwa.Kuonjezera apo, LAMP ndi yoyenera kudziwika kwamitundu yambiri yophatikizidwa mu chip chimodzi, chomwe chili chofunikira kwambiri, komabe pali malo ambiri opangira chitukuko.
Mlingo wabwino kwambiri wa LAMP ukhoza kuchepetsedwa pang'ono ndi RPA, chifukwa kutentha kochepa kwambiri (~ 37 ° C) kumabweretsa zovuta zochepa za evaporation [77].Mu dongosolo la RPA, zoyambira ziwiri zotsutsana zimayambitsa kaphatikizidwe ka DNA pomangiriza ku recombinase ndikukulitsa kumatha mkati mwa mphindi 10 [78,79,80,81].Chifukwa chake, njira yonse ya RPA ndiyothamanga kwambiri kuposa PCR kapena LAMP.M'zaka zaposachedwapa, teknoloji ya microfluidic yasonyezedwa kuti ipititse patsogolo kuthamanga ndi kulondola kwa RPA [82,83,84].Mwachitsanzo, Liu et al.[85] adapanga njira ya microfluidic integrated lateral flow polymerase recombinase amplification assay kuti azindikire mwachangu komanso momveka bwino za SARS-CoV-2 pophatikiza reverse transcript RPA (RT-RPA) ndi njira yodziwira mizere yoyeserera yapadziko lonse lapansi.kulowa mu microfluidic system imodzi.Chithunzi 4b).Malire ozindikira ndi 1 kopi/µl kapena 30 makope/chitsanzo, ndipo kuzindikira kumatha pafupifupi mphindi 30.Kong et al.apanga chipangizo cha microfluidic chovala.[86] adagwiritsa ntchito kutentha kwa thupi komanso njira yodziwira ma fluorescence pafoni yam'manja kuti azindikire mwachangu komanso mwachindunji HIV-1 DNA pogwiritsa ntchito RPA (Chithunzi 4c).Kuyeza kwa RPA komwe kumavala kumazindikira makope 100/mL yazomwe zatsata mkati mwa mphindi 24, kuwonetsa kuthekera kwakukulu kozindikira mwachangu makanda omwe ali ndi kachilombo ka HIV-1 m'malo opanda zida.
Isothermal amplification in point-of-care test (POCT).Kupanga ndi kupanga spin ndi reaction SlipChip.Pambuyo kuwotcherera kwa plasma, tchipisi chapamwamba ndi chapansi chinasonkhanitsidwa ndi mtedza kuti apange chip chomaliza (chosinthidwa kuchokera ku [76]).b Schematic of the MI-IF-RPA system for COVID-19 kuzindikira (yosinthidwa kuchokera [85]).c Schematic ya mayeso ovala a RPA kuti azindikire mwachangu HIV-1 DNA (yosinthidwa kuchokera ku [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 kuwala emitting diode, MI-Flumeral Recifina Akuluakulu Akuluakulu Akuluakulu a LED, MI-fluorescen RecombiteA-FI-Idara Info Kukulitsa
RPA yochokera ku Microfluidic ikukula mwachangu, komabe, mtengo wopangira chip ndikugwiritsa ntchito kachitidwe ndi wokwera kwambiri ndipo uyenera kuchepetsedwa kuti uwonjezere kupezeka kwaukadaulowu.Kuphatikiza apo, kukhudzika kwakukulu kwa RPA kumatha kukhudza kukulitsa kwazinthu zomwe sizili zenizeni, makamaka pamaso pa kuipitsidwa.Zolepheretsa izi zitha kukhudza kugwiritsa ntchito RPA m'makina a microfluidic ndikuyenera kukhathamiritsanso.Zoyambira zopangidwira bwino komanso zofufuzira pazolinga zosiyanasiyana zimafunikiranso kuti athe kukonza njira za RPA-based microfluidic strategy mu POCT.
Cas13 ndi Cas12a amatha kung'amba mwachisawawa ma nucleic acid ndipo motero amatha kupangidwa ngati zida zodziwira komanso zowunikira.Cas13 ndi Cas12a amayatsidwa akamangirira ku DNA kapena RNA, motsatana.Akayatsidwa, puloteniyo imayamba kung'amba ma nucleic acids ena oyandikana nawo, pambuyo pake ma RNA omwe akulozera ma nucleic acid omwe amatha kuphatikizira ma probe ozimitsidwa ndikutulutsa fluorescence.Kutengera chiphunzitsochi, Kellner et al.[87] adapanga njira yochokera ku Cas13 [Specific High-sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK)], ndi Broughton et al.[88] adapanga njira ina yotengera Cas12a [CRISPR Trans Reporter yolunjika ku DNA endonuclease (DTECR)].
M'zaka zaposachedwa, njira zosiyanasiyana zodziwira ma nucleic acid kutengera CRISPR zawoneka [89, 90].Njira zodziwika bwino za CRISPR nthawi zambiri zimawononga nthawi komanso ntchito zambiri chifukwa cha njira zingapo kuphatikiza nucleic acid extraction, amplification ndi CRISPR kuzindikira.Kuwonetsa zamadzimadzi mumlengalenga kungapangitse mwayi wopeza zotsatira zabodza.Potengera zomwe tafotokozazi, machitidwe ozikidwa pa CRISPR akufunika kwambiri kukhathamiritsa.
Pulatifomu ya microfluidic yoyendetsedwa ndi pneumatic yomwe imatha kusanthula 24 mofananira yapangidwira ntchito zowunikira za CRISPR-Cas12a ndi CRISPR-Cas13a [91].Dongosololi lili ndi chipangizo chodziwikiratu cha fluorescence chomwe chimalambalala nucleic acid amplification ndikuzindikira zokha zitsanzo za femtomolar DNA ndi RNA.Chen et al.[92] Integrated recombinase amplification ndi CRISPR-Cas12a dongosolo mu centrifugal microfluidics (Mkuyu 5a).Ntchitoyi imagonjetsa zovuta zophatikiza njira ziwirizi chifukwa Cas12a imatha kukumba DNA ya messenger ndikuletsa njira yokulitsa.Kuphatikiza apo, Chen et al.[92] Komanso chisanadze kusungidwa reagents anachita mu ulamuliro centrifugal microfluidic basi kumaliza ndondomeko yonse.Mu ntchito ina, Silva et al.[93] adapanga njira yodziwira matenda popanda CRISPR / Cas12a amplification ndi foni yamakono kuti azindikire SARS-CoV-2 (Mkuyu 5b).Kuyesa uku, komwe kumadziwika kuti foni-based amplification-free system, kumaphatikizapo enzyme yodalira CRISPR/Cas yomwe imachokera pakuwonetsa ma smartphone a ma siginecha opangidwa ndi catalase mumayendedwe a microfluidic.Kuzindikira kovutirapo kwa makope ochepera 50/µl a nucleic acid popanda kukulitsa, njira yonse kuyambira jekeseni wachitsanzo kupita ku kuwerenga kwazizindikiro kumatenga mphindi 71 zokha.
Njira zodziwira za Nucleic acid zochokera ku CRISPR.Centrifugal POCT yowunikira ma cell ophatikizika otengera CRISPR (yosinthidwa kuchokera ku [92]).b Kukula kwa mayeso a CASCADE pakuwunika kwapa foni yam'manja kwa SARS-CoV-2 (yosinthidwa kuchokera [93]).RAA recombinase amplification, PAM moyandikana ndi protospacer motif, CRISPR yophatikizana kubwereza kwakanthawi kochepa kwa palindromic pafupipafupi, dongosolo la CASCADE lopanda kukulitsa foni yam'manja ndi ma enzymes odalira CRISPR/CAS, 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride EDC
Monga gawo lomaliza la kuzindikira kwa nucleic acid, kuzindikira kwa chizindikiro kumawonetsa zotsatira zowunikira ndipo ndizofunikira kwambiri pakupanga POCT yogwira mtima, yozindikira, komanso yolondola.Zizindikiro zimatha kuwerengedwa pogwiritsa ntchito njira zosiyanasiyana monga fulorosenti, electrochemical, colorimetric ndi maginito.Mu gawo ili, ife kufotokoza zomveka kwa njira iliyonse ndi kuyerekeza diagnostics maselo a matenda opatsirana mu microfluidics.
Njira zozikidwa pa fluorescence zimagwiritsidwa ntchito kwambiri pakuwunika kwa POCT kwa matenda opatsirana chifukwa cha zabwino zake zodziwika bwino, zotsika mtengo, zosavuta kugwira ntchito, komanso kusanthula kwachisamaliro [94, 95].Njirazi zimagwiritsa ntchito ma fluorophores olembedwa ngati utoto wa fulorosenti ndi nanomaterials kuti apange chizindikiro chodziwika (fluorescence enhancement or quenching).Zomwe anapezazi zikusonyeza kuti njira zogwiritsira ntchito fulorosenti zitha kugawidwa m'malembo achindunji a fulorosenti, kuyatsa, ndi kuzindikira kwa fluorescent [96].Kuzindikira label ya fluorescent molunjika kumagwiritsa ntchito zilembo zapadera za fulorosenti kuti zilembe ma ligand omwe amapanga kuchuluka kwa fulorosenti akamangika ku chandamale.Kuti muzindikire zamtundu wa fluorescence, mtundu wa chizindikiro cha fulorosenti umagwirizana bwino ndi kukula kwa chidwi.Kuchuluka kwa Fluorescence sikungatheke popanda chandamale ndipo kumawoneka ngati chandamale chokwanira chilipo.Mosiyana ndi zimenezi, mphamvu ya fluorescence yodziwika ndi "signal-off" fluorescence imagwirizana mosagwirizana ndi chiwerengero cha chandamale, poyambira kufika pamtengo wapatali ndipo pang'onopang'ono imachepa pamene cholingacho chikukulitsidwa.Mwachitsanzo, pogwiritsa ntchito njira ya CRISPR-Cas13a yodalira trans-cleavage, Tian et al.[97] adapanga njira yodziwikiratu kuti azindikire ma RNA omwe amadumpha modumphadumpha molunjika (mkuyu 6a).Mukamangiriza ku ma RNAs owonjezera, CRISPR-Cas13-RNA zovuta zitha kuyambitsidwa, ndikuyambitsa kupatukana ndi mtolankhani wosadziwika wa RNAs.Mtolankhani wolembedwa ndi fluorescently [fluorophore (F)] amazimitsidwa ndi chozimitsa (Q) chosasunthika komanso ma fluoresces akamang'ambika ndi zovuta zomwe zidalowetsedwa.
Ubwino wa kuzindikira kwa electrochemical ndikuthamanga kwambiri, kupanga kosavuta, mtengo wotsika, kosavuta kunyamula komanso kuwongolera zokha.Ndi njira yamphamvu yowunikira pamapulogalamu a POCT.Kutengera graphene field-effect transistors Gao et al.[98] adapanga nanobiosensor kwa multiplex kuzindikira ma antigen a matenda a Lyme kuchokera ku mabakiteriya a Borrelia burgdorferi okhala ndi malire a 2 pg / mL (Mkuyu 6b).
Zoyesa zamtundu zakhala zikugwiritsidwa ntchito mu mapulogalamu a POCT, kupindula ndi zabwino za kunyamula, mtengo wotsika, kukonzekera kosavuta, komanso kuwerenga kowoneka.Kuzindikira kwamtundu kumatha kugwiritsa ntchito okosijeni wa peroxidase kapena peroxidase-like nanomaterials, kuphatikiza kwa nanomaterials, komanso kuwonjezera utoto wamtundu kuti asinthe zambiri za kukhalapo kwa chandamale cha nucleic acid kukhala kusintha kowoneka bwino kwamitundu [99, 100, 101].Makamaka, ma nanoparticles agolide amagwiritsidwa ntchito kwambiri popanga njira zamitundu, ndipo chifukwa cha kuthekera kwawo kupangitsa kusintha kwamitundu mwachangu komanso kofunikira, pali chidwi chochulukirapo pakupanga nsanja zamtundu wa POCT zowunikira matenda opatsirana [102].Ndi chipangizo chophatikizika cha centrifugal microfluidic [103], tizilombo toyambitsa matenda m'zakudya zamkaka zowonongeka zimatha kudziwika pamlingo wa maselo a bakiteriya a 10, ndipo zotsatira zake zikhoza kuwerengedwa mowonekera mkati mwa mphindi 65 (mkuyu 6c).
Njira zodziwira maginito zimatha kuzindikira zowunikira pogwiritsa ntchito maginito, ndipo pakhala chidwi chachikulu pakugwiritsa ntchito kwa POCT m'zaka zaposachedwa.Njira zowonera maginito zili ndi zabwino zina zapadera monga zida zotsika mtengo za maginito m'malo mwa zida zowoneka bwino.Komabe, kugwiritsa ntchito maginito kumathandizira kuzindikira komanso kumachepetsa nthawi yokonzekera [104].Kuphatikiza apo, zotsatira za kufufuza kwa maginito zikuwonetsa kutsimikizika kwakukulu, kukhudzika, komanso chiŵerengero chapamwamba cha maginito ndi phokoso chifukwa cha chizindikiro chochepa cha maginito cha zitsanzo zamoyo [105].Sharma et al.adaphatikizira njira yolumikizira maginito yochokera ku biosensor kukhala nsanja yonyamula ya microchip.[106] pofuna kudziwa zambiri za tizilombo toyambitsa matenda (mkuyu 6d).Ma biosensors amazindikira mosamala ma subnanomolar nucleic acid otalikirana ndi tizilombo toyambitsa matenda.
Njira yodziwira chizindikiro.Lingaliro la kuzindikira kwa hyperlocalized kwa Cas13a (losinthidwa kuchokera ku [97]).b Graphene nanobiosensor FET kuphatikiza Lyme GroES scFv (yosinthidwa kuchokera ku [98]).c Colorimetric zizindikiro kwa multiplex kudziwa tizilombo toyambitsa matenda chakudya mu centrifugal microfluidic chip: No. 1 ndi No. 3 zitsanzo ndi chandamale tizilombo toyambitsa matenda, ndi No. 2, No. 4 ndi No. .d Biosensor yochokera pamphambano yamaginito, kuphatikiza nsanja, chotchingira chotchinga chomangirira, gawo lowongolera, ndi magetsi opanga ma siginecha / kupeza (zosinthidwa kuchokera ku [106]).GFET Graphene FET, Escherichia coli, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus, Vibrio parahaemolyticus, Listeria monocytogenes, PC PC, PDMS Dimethicone, PMMA polymethyl methacrylate
Ngakhale kuti ali ndi makhalidwe abwino kwambiri a njira zodziwira pamwambazi, ali ndi zovuta zake.Njirazi zimafaniziridwa (tebulo 1), kuphatikiza mapulogalamu ena okhala ndi tsatanetsatane (zabwino ndi zoyipa).
Ndi chitukuko cha microfluidics, microelectromechanical systems, nanotechnology ndi sayansi ya zipangizo, kugwiritsa ntchito tchipisi tating'onoting'ono tomwe timayang'ana matenda opatsirana kumapita patsogolo [55,96,107,108].Kuwongolera molondola kwa zida zazing'ono ndi zamadzimadzi kumathandizira kuti zidziwitso zikhale zolondola komanso zotsika mtengo.Chifukwa chake, kuti apititse patsogolo, kuyesayesa kwapangidwa kukhathamiritsa ndikukweza tchipisi, zomwe zimapangitsa kuti pakhale tchipisi tating'ono tating'ono tating'ono tating'ono tating'ono tating'ono ndi magwiridwe antchito osiyanasiyana.Apa tikuwonetsa mwachidule mitundu ingapo yodziwika bwino yamapulatifomu a microfluidic ndikuyerekeza mawonekedwe awo (zabwino ndi zoyipa).Kuphatikiza apo, zambiri mwazitsanzo zomwe zalembedwa pansipa zimayang'ana kwambiri polimbana ndi SARS-CoV-2.
LOCCs ndizomwe zimagwiritsidwa ntchito kwambiri ndi miniaturized complex analytical systems ndipo ntchito zawo zimakhala zochepa kwambiri, zophatikizika, zowonongeka komanso zofanana kuchokera ku jekeseni wa zitsanzo ndi kukonzekera, kuyendetsa kayendedwe ka madzi ndi kuzindikira kwamadzimadzi [109, 110].Zamadzimadzi zimasinthidwa kudzera mu geometry yopangidwa mwaluso komanso kuyanjana kwazinthu zambiri zakuthupi monga ma pressure gradient, capillary action, electrodynamics, magnetic fields and acoustic wave [111].LOCC ikuwonetsa zabwino kwambiri pakuwunika kwapamwamba kwambiri komanso kuzindikira kangapo, ndikufulumira kusanthula, kukula kwachitsanzo chaching'ono, kugwiritsa ntchito mphamvu zochepa, komanso kasamalidwe kapamwamba komanso magwiridwe antchito;komabe, zida za LOCC ndizovuta kwambiri, ndikupanga, kulongedza, komanso kulumikizana.Komabe, kuchulukitsa ndikugwiritsanso ntchito kumakumana ndi zovuta zazikulu [96].Poyerekeza ndi nsanja zina, LOCC ili ndi maubwino apadera potengera kusiyanasiyana kwa magwiridwe antchito komanso ukadaulo wabwino kwambiri, koma zovuta zake ndizodziwikiratu, zomwe ndizovuta kwambiri komanso kusabwerezabwereza.Kudalira mapampu akunja, omwe nthawi zambiri amakhala ochulukirapo komanso okwera mtengo, amalepheretsanso kugwiritsidwa ntchito kwawo mu POCT.
Panthawi ya mliri wa COVID-19, LOCC idalandira chidwi kwambiri.Nthawi yomweyo, pali tchipisi tatsopano tomwe timaphatikiza matekinoloje angapo.Mwachitsanzo, mafoni a m'manja tsopano amagwiritsidwa ntchito kwambiri ngati zida zowerengera ndipo ali ndi kuthekera kwakukulu kophatikiza LOCC.Sun et al.[21] adapanga chip cha microfluidic chomwe chimalola kuti ma nucleic acid achuluke machulukidwe asanu a tizilombo toyambitsa matenda, kuphatikiza SARS-CoV-2, pogwiritsa ntchito LAMP ndikusanthula pogwiritsa ntchito foni yamakono mkati mwa ola la 1 pambuyo pomaliza.Monga chitsanzo china, Sundah et al.[112] adapanga masinthidwe a mamolekyulu [kukulitsa chothandizira ndi mamolekyulu osintha ma cell (CATCH)] kuti azindikire molunjika komanso momveka bwino za zolinga za SARS-CoV-2 RNA pogwiritsa ntchito mafoni. CATCH imagwirizana ndi LOCC yonyamula ndipo imakwaniritsa bwino kwambiri (pafupifupi makope a 8 RNA / μl; <1 h kutentha kwachipinda) [112]. CATCH imagwirizana ndi LOCC yonyamula ndipo imakwaniritsa bwino kwambiri (pafupifupi makope a 8 RNA / μl; <1 h kutentha kwachipinda) [112]. CATCH совместим с портативным LOCC ndi обеспечивает превосходную производительность (примерно 8 копий РНК/мкл; <1 ч при компечи12]. CATCH imagwirizana ndi LOCC yonyamulika ndipo imapereka njira yabwino kwambiri (pafupifupi makope 8 a RNA/µl; <1 h kutentha kwachipinda) [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 ndi обладает превосходной производительностью (примерно 8 копий РНК/мкл; <1 часа при комнат121 тем) [ CATCH imagwirizana ndi ma LOCC osunthika ndipo imagwira ntchito bwino kwambiri (pafupifupi makope 8 a RNA/µl; <ola 1 pa kutentha kwachipinda) [112].Kuphatikiza apo, zida za LOCC zowunikira ma cell zimagwiritsanso ntchito mphamvu zina zoyendetsa monga vacuum, kutambasula, ndi minda yamagetsi.Kang et al.[113] adawonetsa nthawi yeniyeni, yothamanga kwambiri ya nanoplasma-on-a-chip PCR yowunikira mwachangu komanso mochulukira wa COVID-19 m'munda pogwiritsa ntchito vacuum plasmonic liquid PCR chip.Li et al.[114] pambuyo pake adapanga chipangizo chowongolera chowongolera cha microfluidic chomwe chidathandizira kuzindikira COVID-19.Pulatifomu imagwiritsa ntchito RT-LAMP amplification system kuti idziwe ngati chitsanzo chili chabwino kapena cholakwika.Pambuyo pake, Ramachandran et al.[115] adapeza ma gradients oyenerera amagetsi pogwiritsa ntchito isotachophoresis (ITP), njira yosankha ya ion yomwe imagwiritsidwa ntchito mu microfluidics.Ndi ITP, chandamale cha RNA kuchokera ku zitsanzo za swab za nasopharyngeal zitha kuyeretsedwa zokha.Ndiye Ramachandran et al.[115] Kuphatikiza kuyeretsedwa kwa ITP uku ndi zoyeserera za ITP-zowonjezera za LAMP ndi CRISPR zidazindikira SARS-CoV-2 mu swab ya nasopharyngeal yamunthu ndi zitsanzo zachipatala pafupifupi mphindi 35.Kuonjezera apo, malingaliro atsopano akutuluka nthawi zonse.Jadhav et al.[116] anakonza njira yodziwira matenda pogwiritsa ntchito mawonekedwe apamwamba a Raman spectroscopy pamodzi ndi chipangizo cha microfluidic chokhala ndi golide / siliva-wokutidwa ndi carbon nanotubes kapena ma electrospun micro/nanotubes otayika.Ma membrane-functionalized build-in fltering microchannels amatha kutaya.Chipangizochi chimatsatsa ma virus kuchokera kumadzimadzi osiyanasiyana amthupi / ma exudations monga malovu, nasopharynx ndi misozi.Chifukwa chake, kachilombo ka HIV kachuluka ndipo kachilomboka kamatha kudziwika bwino ndi siginecha ya Raman.
LOAD ndi nsanja ya centrifugal microfluidic momwe njira zonse zimayendetsedwa ndi ma frequency protocol omwe amazungulira gawo laling'ono lopangidwa ndi microstructured [110].Chipangizo cha LOAD chimadziwika pogwiritsa ntchito mphamvu ya centrifugal ngati mphamvu yoyendetsa galimoto.Zamadzimadzi zimakhalanso ndi mphamvu za capillary, Euler ndi Coriolis.Pogwiritsa ntchito chipangizo cha centrifuge, kusanthula kumachitika mosalekeza pakugwira ntchito kwamadzimadzi kuchokera ku radial kulowa mkati kupita kumalo akunja, kuchotsa kufunikira kwa machubu owonjezera akunja, mapampu, ma actuators, ndi ma valve ogwira ntchito.Mwachidule, njira imodzi yowongolera imathandizira kugwira ntchito.Mphamvu zomwe zimagwira pamadzi mu njira yofanana ya microfluidic pamtunda wofanana ndi malo olemetsa ndi ofanana, zomwe zimapangitsa kuti zitheke kubwereza ndondomekoyi.Chifukwa chake, zida za LOAD ndizosavuta komanso zotsika mtengo kupanga ndi kupanga kuposa zida wamba za LOCC, pomwe machitidwewo amakhala odziyimira pawokha komanso ofanana;komabe, chifukwa cha mphamvu zamakina apamwamba a zida za centrifugal, zida za chip zomwe zilipo ndizochepa ndipo ma voliyumu ang'onoang'ono ndi ovuta.ku galimoto.Panthawi imodzimodziyo, zipangizo zambiri za LOAD zimapangidwira ntchito imodzi yokha, zomwe zimakhala zokwera mtengo kuti zizindikire zazikulu [96, 117, 118, 119].
M'zaka zaposachedwa, LOAD, yomwe imadziwika kuti ndi imodzi mwazinthu zodalirika kwambiri za microfluidic, yalandira chidwi kwambiri kuchokera kwa ofufuza ndi opanga.Chifukwa chake, LOAD yalandiridwa kwambiri ndipo yakhala ikugwiritsidwa ntchito pozindikira ma cell a tizilombo toyambitsa matenda [120, 121, 122, 123, 124], makamaka pakubuka kwa COVID-19.Mwachitsanzo, kumapeto kwa 2020, Ji et al.[60] adawonetsa kuyesa kwachindunji kwa RT-qPCR pakuzindikira mwachangu komanso kodziwikiratu kwa SARS-CoV-2 ndi matenda a fuluwenza A ndi B pazitsanzo zapakhosi.Ndiye Xiong et al.[74] adapereka nsanja ya LAMP-integrated discoid microfluidic kuti azindikire mwachangu, molondola, komanso munthawi yomweyo ma virus asanu ndi awiri opuma a anthu, kuphatikiza SARS-CoV-2, mkati mwa mphindi 40.Kumayambiriro kwa 2021, de Oliveira et al.[73] adawonetsa chip cha polystyrene tona centrifugal microfluidic, chogwiritsidwa ntchito pamanja ndi chozungulira chala chala, pozindikira matenda a RT-LAMP a COVID-19.Pambuyo pake, Dignan et al.[39] adapereka chida chodzitchinjiriza cha centrifuge choyeretsera SARS-CoV-2 RNA mwachindunji kuchokera ku magawo a buccal swab.Medved et al.[53] anaganiza zokhala ndi inline SARS-CoV-2 aerosol sampling system yokhala ndi chip yaying'ono yozungulira ya microfluidic fulorescent yokhala ndi malire ozindikira makope 10/μL ndi gawo locheperako la mphindi 15.Suarez et al.[75] Posachedwapa linanena za chitukuko cha Integrated modular centrifugal microfluidic nsanja kuti azindikire mwachindunji SARS-CoV-2 RNA mu kutentha-inactivated nasopharyngeal swab zitsanzo ntchito LAMP.Zitsanzo izi zikuwonetsa maubwino ndi lonjezo la LOAD pakuwunika kwa ma cell a COVID-19.
Mu 1945 Muller ndi Clegg [125] adawonetsa koyamba njira za microfluidic pamapepala pogwiritsa ntchito pepala losefera ndi parafini.Mu 2007, gulu la Whitesides [126] lidapanga nsanja yoyamba yogwira ntchito yoyesa mapuloteni ndi shuga.Pepala lakhala gawo lapansi labwino la microfluidics.Pepalali lili ndi zinthu zachilengedwe monga hydrophilicity ndi porous structure, biocompatibility yabwino, kulemera kopepuka, kusinthasintha, kupindika, mtengo wotsika, kugwiritsa ntchito mosavuta komanso kosavuta.Ma µPAD akale amakhala ndi zinthu za hydrophilic/hydrophobic zomangidwa pamapepala.Kutengera ndi mawonekedwe amitundu itatu, ma μPAD amatha kugawidwa m'magulu awiri (2D) ndi atatu-dimensional (3D) μPAD.2D µPADs amapangidwa popanga malire a hydrophobic kuti apange mayendedwe a microfluidic, pomwe 3D µPADs nthawi zambiri amapangidwa kuchokera kumagulu a mapepala a 2D microfluidic, nthawi zina ndi mapepala, njira zozembera, njira zotseguka, ndi kusindikiza kwa 3D [96].Madzi amadzimadzi kapena achilengedwe pa μPAD amayendetsedwa makamaka ndi mphamvu ya capillary popanda gwero lamphamvu lakunja, kuthandizira kusungiratu kusungirako kwa reagents, kasamalidwe ka zitsanzo, ndi kuzindikira ma multiplex.Komabe, kuwongolera kolondola komanso kuzindikira kwa ma multiplex kumalepheretsedwa ndi kuthamanga kosakwanira kuzindikira, kukhudzika, komanso kusinthikanso [96, 127, 128, 129, 130].
Monga nsanja yachilendo ya microfluidic, μPAD yakhala ikukwezedwa kwambiri ndikupangidwa kuti izindikire matenda opatsirana monga HCV, HIV, ndi SARS-CoV-2 [131, 132].Posankha komanso kuzindikira kwa HCV, Tengam et al.[133] adapanga buku la biosensor lotengera pepala la fulorosenti pogwiritsa ntchito kafukufuku wodziwika bwino wa nucleic acid wozikidwa pa pyrrolidinyl peptide.Ma Nucleic acids amakhala osasunthika pamapepala a cellulose omwe ali ndi okosijeni pang'ono pochepetsa kuphatikizika pakati pamagulu amino ndi magulu a aldehyde, ndipo kuzindikira kumatengera fluorescence.Zizindikirozi zimatha kuwerengedwa ndi chida chopangidwa mwapadera chokhala ndi kamera yonyamula fulorosenti kuphatikiza ndi kamera ya foni yam'manja.Pambuyo pake, Lu et al.[134] adapanga ma elekitirodi osinthika pamapepala otengera faifi tambala/golide nanoparticles/carbon nanotubes/polyvinyl alcohol organometallic framework composites kuti azindikire chandamale cha HIV ndi DNA hybridization pogwiritsa ntchito methylene buluu ngati chizindikiro cha DNA redox.Posachedwapa, Chowdury et al.[135] adapereka mawonekedwe ongoyerekeza a pulatifomu yoyeserera µPAD pogwiritsa ntchito malovu osaphika a odwala kuphatikiza ndi LAMP ndi ukadaulo wojambula wonyamula kuti azindikire analyte wa COVID-19.
Mayeso oyenda pambuyo pake amawongolera zamadzimadzi pogwiritsa ntchito mphamvu ya capillary ndikuwongolera kusuntha kwamadzimadzi potengera kunyowa komanso mawonekedwe a ma porous kapena microstructured substrates.Zida zoyendera kutsogolo zimakhala ndi zitsanzo, conjugate, incubator ndi kuzindikira, ndi mapepala otsekemera.Ma nucleic acid mu LFA amazindikira zomangira zina zomwe zimasungidwa pamalo omangira ndikumanga ngati zovuta.Pamene madzi akudutsa muzitsulo zopangira ma incubation ndi kuzindikira, zovutazo zimagwidwa ndi mamolekyu ogwidwa omwe ali pa mizere yoyesera ndi kulamulira, kusonyeza zotsatira zomwe zingathe kuwerengedwa mwachindunji ndi maso.Nthawi zambiri, LFA imatha kumalizidwa mu mphindi 2-15, zomwe zimathamanga kuposa zomwe zidapezeka kale.Chifukwa cha makina apadera, LFA imafuna ntchito zochepa ndipo sichifuna zipangizo zowonjezera, zomwe zimapangitsa kuti zikhale zosavuta kugwiritsa ntchito.Ndi zophweka kupanga ndi miniaturize, ndipo mtengo wa magawo opangidwa ndi mapepala ndi wotsika.Komabe, zimangogwiritsidwa ntchito pofufuza zamtengo wapatali, ndipo kudziwa kuchuluka kwake kumakhala kovuta kwambiri, ndipo kuthekera kochulukitsa ndi kupititsa patsogolo kumakhala kochepa kwambiri, ndipo nucleic acid imodzi yokha yokwanira imatha kupezeka panthawi imodzi [96,110,127].
Ngakhale kuti ntchito zambiri za LFA zimangoyang'ana ma immunoassays, kugwiritsa ntchito LFA pakuzindikira ma cell mu tchipisi tating'onoting'ono kumakhala kothandiza komanso kotchuka [136].Pankhani ya kachilombo ka hepatitis B, HIV ndi SARS-CoV-2 LFA Gong et al.[137] anakonza nsanja ya nanoparticle LFA yosinthira ndikuwonetsa kusinthasintha kwa nsanja yaying'ono komanso yosunthika kudzera pakuzindikira kovutirapo komanso kochulukira kwa zolinga zingapo monga HBV nucleic acid.Kuphatikiza apo, Fu et al.[138] adawonetsa buku la LFA lotengera mawonekedwe owoneka bwino a Raman pakuwunika kuchuluka kwa HIV-1 DNA pazambiri zotsika.Kuti muzindikire mwachangu komanso movutikira kwa SARS-CoV-2, Liu et al.[85] adapanga kafukufuku wa microfluidic-integrated RPA lateral flow flow pophatikiza RT-RPA ndi njira yodziwikiratu yapadziko lonse lapansi munjira imodzi ya microfluidic.
Kugwiritsa ntchito nsanja zosiyanasiyana za microfluidic kumasiyanasiyana kutengera maphunziro enaake, kugwiritsa ntchito bwino mphamvu ndi zabwino zamapulatifomu.Ndi ma valve otsika mtengo, mapampu ndi ma ducts, LOCC ndiye nsanja yokwanira kwambiri yogwiritsira ntchito zosiyanasiyana komanso kugwirizanirana ndi chipinda chachikulu kwambiri chachitukuko.Chifukwa chake, tikuyembekeza ndikupangira kuti maphunziro aposachedwa kwambiri achitidwe ku LOCC ngati kuyesa koyamba ndikuti mikhalidweyo ikonzedwe.Kuonjezera apo, njira zowonjezereka komanso zolondola zikuyembekezeka kupezedwa ndikugwiritsidwa ntchito mu dongosolo.LOAD imapambana pakuwongolera bwino kwamadzi kuchokera kuzipangizo zomwe zilipo kale za LOCC ndipo imawonetsa maubwino apadera pamayendedwe amodzi ndi mphamvu yapakati popanda kufunikira kwa ma drive akunja, pomwe mayankho ofanana amatha kukhala osiyana ndi kulumikizidwa.Chifukwa chake, m'tsogolomu, LOAD idzakhala nsanja yayikulu ya microfluidic yokhala ndi ntchito zochepa pamanja komanso umisiri wokhwima komanso wodzipangira.Pulatifomu ya µPAD imaphatikiza phindu la LOCC ndi zida zopangira mapepala zotsika mtengo, zowunika kugwiritsa ntchito kamodzi.Chifukwa chake, chitukuko chamtsogolo chiyenera kuyang'ana pa matekinoloje osavuta komanso okhazikitsidwa bwino.Kuonjezera apo, LFA ndi yoyenera kuyang'ana maso amaliseche, ndikulonjeza kuchepetsa kugwiritsira ntchito zitsanzo ndikufulumira kuzindikira.Kuyerekeza kwatsatanetsatane kwa nsanja kukuwonetsedwa mu Gulu 2.
Kusanthula kwa digito kumagawanitsa zitsanzozo kukhala ma microreactors ambiri, omwe ali ndi mamolekyu angapo omwe amawatsata [139, 140].Kuyeza kwapa digito kumapereka zabwino zambiri poyesa kuchuluka kwathunthu mwa kuyesa masauzande ambiri ofanana a biochemical panthawi imodzi komanso payekhapayekha m'magawo a micron osati mopitilira.Poyerekeza ndi ma microfluidic achikhalidwe, machitidwe a chipinda amatha kuchepetsa kuchuluka kwa zitsanzo, kukulitsa magwiridwe antchito, ndikuphatikizidwa mosavuta ndi njira zina zowunikira popanda kufunikira kwa njira, mapampu, mavavu, ndi mapangidwe ophatikizika [141, 142, 143, 144, 145, 146, 147] .Njira ziwiri zotsatirazi zimagwiritsidwa ntchito poyesa digito kuti akwaniritse kulekanitsa yunifolomu komanso yolondola ya mayankho, kuphatikizapo ma reagents ndi zitsanzo monga maselo, nucleic acid, ndi particles kapena mamolekyu ena: (1) dontho emulsions kugwiritsa ntchito kusakhazikika kwa mawonekedwe a madzi;(2) kugawidwa kwamagulu kumachitika ndi zovuta za geometric za chipangizocho.Mu njira yoyamba, madontho okhala ndi ma reagents ndi zitsanzo mu ma microchannels amatha kupangidwa ndi njira zopanda pake monga co-current, crossflow, flow focusing, stage emulsification, microchannel emulsification, ndi nembanemba kupyolera mu mphamvu za viscous shear ndi emulsification ndi kusintha kwa njira.localization [143, 145, 146, 148, 149] kapena kugwiritsa ntchito njira zogwira ntchito [150, 151], zomwe zimayambitsa mphamvu zowonjezera kudzera mu mphamvu zamagetsi, maginito, kutentha ndi makina.M'njira yomalizayi, kufanana kwamadzimadzi kwabwino kwambiri m'zipinda za microfluidic kumagawidwa mwa kusunga malo amtundu wofanana, monga ma micropits ndi pamwamba [152,153,154].Makamaka, madontho ndi zigawo zazikulu zoyenda zomwe zimatha kupangidwanso ndikusinthidwa pama electrode arrays kutengera digito microfluidics (DMF).Electrowetting of dielectrics ndi imodzi mwazambiri zophunziridwa bwino kwambiri za DMF, popeza kulowetsedwa kwa ma dielectrics kumalola kuwongolera bwino madontho amtundu uliwonse, kuwongolera mawonekedwe amadzimadzi ndi ma siginecha amagetsi akudutsa mbali zosiyanasiyana [141, 144].Ntchito zazikulu zokhala ndi madontho mu DMF zimaphatikizapo kusanja, kupatukana, ndi kuphatikiza [151, 155, 156], zomwe zingagwiritsidwe ntchito m'magawo osiyanasiyana owunikira, makamaka pozindikira maselo [157, 158, 159].
Kuzindikira kwa Digital nucleic acid ndiukadaulo wam'badwo wachitatu wowunikira ma cell omwe amatsatira PCR wamba komanso kuchuluka kwa PCR (qPCR), motsatana ndi kutsatizana kwakukulu komanso kufufuza kwamadzimadzi.M'zaka makumi awiri zapitazi, ma nucleic acids a digito adakula mwachangu pantchito yowunikira ma cell a tizilombo toyambitsa matenda [160, 161, 162].Kuchulukitsidwa kokwanira kwa kuzindikira kwa digito ya nucleic acid kumayamba ndikulongedza zitsanzo ndi ma reagents m'zigawo zapagulu kuti zitsimikizire kuti mndandanda uliwonse wa chandamale uli ndi mwayi wofanana wolowa m'chipinda chilichonse.Mwachidziwitso, gawo lirilonse likhoza kupatsidwa maulendo angapo, kapena sipangakhale makina odziimira okhaokha.Kupyolera mu njira zosiyanasiyana zodziwira zomwe tafotokozazi, zipinda zokhala ndi ma tambala ang'onoang'ono omwe amapanga zizindikiro pamwamba pa malo enaake amatha kuwonedwa ndi maso kapena makina ndipo amalembedwa ngati zabwino, pamene zipinda zina zomwe zimapanga zizindikiro pansi pa khomo zimatchedwa zabwino. .zoipa, zomwe zimapangitsa chizindikiro cha gawo lililonse kukhala boolean.Chifukwa chake, powerengera kuchuluka kwa zigawo zomwe zidapangidwa komanso kuchuluka kwa zotsatira zabwino pambuyo pa zomwe zidachitika, zolemba zoyambirira zoyeserera zitha kufananizidwa pogwiritsa ntchito njira yogawa ya Poisson popanda kufunikira kokhotakhota kokhazikika, komwe kumafunikira pakuwunika kochulukira kwanthawi zonse. ndi qPCR.[163] Poyerekeza ndi njira zachikhalidwe zodziwira mamolekyulu, kuzindikira kwa digito kwa nucleic acid kumakhala ndi digirii yapamwamba yodzipangira zokha, kuthamanga kwambiri kusanthula ndi kukhudzidwa, ma reagents ochepa, kuipitsidwa pang'ono, komanso kupanga ndi kupanga kosavuta.Pazifukwa izi, kugwiritsa ntchito zoyesa za digito, makamaka njira zotsika, zowunikira ma cell, kuphatikiza kukulitsa ndi njira zowerengera ma siginecha, kwaphunziridwa bwino pakuphulika kwakukulu kwa SARS-CoV-2.Mwachitsanzo, Yin et al.[164] njira zophatikizira zamadontho a digito ndi othamanga a PCR kuti azindikire mitundu ya ORF1ab, N, ndi RNase P mu SARS-CoV-2 mu chip microfluidic.Mwachidziwitso, dongosololi linatha kuzindikira chizindikiro chabwino mkati mwa masekondi a 115, omwe ali mofulumira kuposa PCR wamba, kusonyeza mphamvu yake pozindikira mfundo za chisamaliro (Chithunzi 7a).Dong et al.[165], Bzalani et al.[157], Chen et al.[166] ndi Alteri et al.[167] adagwiritsanso ntchito droplet digital PCR (ddPCR) kuti azindikire SARS-CoV-2 mu microfluidic system yokhala ndi zotsatira zochititsa chidwi.Kuti apititse patsogolo kuchuluka kwa chidziwitso, Shen et al.[168] anapeza ddPCR-based chip imaging mu 15 s popanda kugwiritsa ntchito njira zosokera zithunzi, kufulumizitsa ndondomeko ya teknoloji ya ddPCR kuchokera ku labu kupita ku ntchito.Sikuti njira zokulitsa matenthedwe monga PCR zimagwiritsidwanso ntchito, komanso njira zokulirapo za isothermal zimagwiritsidwa ntchito kuti zifewetse zomwe zimachitika komanso kuyankha mwachangu.Lu et al.[71] adapanga SlipChip yowunikira madontho, otha kutulutsa madontho akulu akulu mosiyanasiyana mu sitepe imodzi ndikuwerengera ma SARS-CoV-2 nucleic acids pogwiritsa ntchito LAMP ya digito (Chithunzi 7b).Monga ukadaulo wosinthika mwachangu, CRISPR imathanso kutenga gawo lofunikira pakuzindikirika kwa digito ya nucleic acid kudzera pazithunzithunzi zosavuta za colorimetric popanda kufunikira kwa madontho owonjezera a nucleic acid.Ackerman ndi al.adapanga combinatorial matrix reaction pakuwunika kwa ma nucleic acid.[158] anapeza mavairasi 169 okhudzana ndi anthu, kuphatikizapo SARS-CoV-2, m'malovu omwe ali ndi CRISPR-Cas13-based nucleic acid reagents mu microwell assay (Chithunzi 7c).Kuphatikiza apo, isothermal amplification ndi CRISPR teknoloji ingagwiritsidwe ntchito mu dongosolo lomwelo kuphatikiza ubwino wa onse awiri.Park et al.[169] Kuyesa kwa digito kwa CRISPR/Cas12a kudapangidwa mu chipangizo cha microfluidic chamalonda kuti chizindikirike cha SARS-CoV-2 yochotsedwa ndi kupha kutentha kutengera gawo limodzi la RT-RPA yokhala ndi chidziwitso chachifupi komanso chapamwamba choyambira kumbuyo. chiŵerengero cha nthawi., kusinthasintha kwakukulu komanso kukhudzidwa bwino (mkuyu 7d).Zofotokozera zina za zitsanzozi zaperekedwa mu Gulu 3.
Pulatifomu ya digito yodziwika bwino ya nucleic acid.a Kuyenda mwachangu kwa digito ya PCR kumakhala ndi njira zinayi zofunika: kukonzekera kwachitsanzo, kugawa zomwe zimasakanikirana, kukulitsa, ndi kuchuluka kwa chandamale (kuchokera ku [164]).b Schematic yowonetsa kusanthula kwa madontho a SlipChip kuti apange madontho pakuchulukira kwakukulu (kusinthidwa kuchokera ku [71]).c CARMEN-Cas chithunzi chakuyenda kwa ntchito13 (chosinthidwa kuchokera [158]).d Mwachidule pakuzindikira kwa kachilombo ka digito ndi CRISPR/Cas mumphika umodzi (wosinthidwa kuchokera ku [169]).W/O madzi-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, kwambiri pachimake kupuma matenda, coronavirus 2 , RT Kukulitsa kwa reverse transcriptase recombinase polymerase-RPA, chizindikiro cha S/B chakumbuyo
Nthawi yotumiza: Sep-15-2022
