Pono nā hoʻolālā diagnostic kuʻuna no ka ʻike ʻana i nā maʻi infectious i ka hoʻohana ʻana i nā mea hana benchtop i kūpono ʻole no ka hoʻāʻo ʻana i ka point-of-care testing (POCT).ʻO ka microfluidics e puka mai nei he ʻenehana liʻiliʻi loa, ʻokoʻa, a i hoʻohui ʻia he mea hiki ke hoʻololi i nā ʻano kuʻuna no ka wikiwiki, haʻahaʻa, pololei ma ka pūnaewele diagnostics.Hoʻohana nui ʻia nā ʻano diagnostic molecular i nā mea microfluidic e like me nā ala kūpono loa no ka ʻike pathogen.Hōʻuluʻulu kēia loiloi i nā holomua hou i ka microfluidic-based molecular diagnostics o nā maʻi infectious mai ka ʻike kula a me ka ʻoihana.ʻO ka mea mua, wehewehe mākou i ka hana maʻamau o nā ʻakika nucleic, me ka hoʻomaʻamaʻa mua ʻana, ka hoʻonui ʻana, a me ka heluhelu hōʻailona.Hoʻohālikelike ʻia nā ʻano, nā pono a me nā pōʻino o nā ʻano microfluidic ʻehā.Ma hope aʻe, e kūkākūkā mākou i ka hoʻohana ʻana i nā hōʻike kikohoʻe no ka helu piha ʻana o nā waikawa nucleic.Ua hōʻuluʻulu ʻia nā ʻenehana diagnostic microfluidic-based ʻelua a me nā ʻoihana hou e like me ka hōʻike o ke kūlana o ka mākeke.ʻO ka hope, ke hāpai nei mākou i nā kuhikuhi i ka wā e hiki mai ana no ka ʻike microfluidic o nā maʻi maʻi.
Hoʻokumu ʻia nā maʻi maʻi e nā pathogens, e komo pū ana me nā maʻi bacteria, nā maʻi maʻi, a me nā parasites, i laha ʻia ma ka honua holoʻokoʻa.ʻAʻole e like me nā maʻi ʻē aʻe, lilo koke nā pathogens i ka maʻi a hoʻolaha i waena o nā kānaka a me nā holoholona hoʻokipa ma o ka inoculation, ka ea a me ka media wai [1].He mea koʻikoʻi ka pale ʻana i ka maʻi maʻi ma ke ʻano he ʻano olakino olakino.ʻEkolu mau kumu nui no ka hakakā ʻana i nā maʻi lele: (1) hoʻomalu i ke kumu o ka maʻi;(2) keakea o ke ala lawe;(3) ka pale ʻana i nā poʻe maʻalahi.Ma waena o nā hoʻolālā nui, ʻo ka mālama ʻana i ke kumu o ka maʻi i manaʻo ʻia ʻo ia ka hoʻolālā koʻikoʻi ma muli o kona ʻoluʻolu a me ke kumukūʻai haʻahaʻa.ʻO ka maʻi wikiwiki, ka hoʻokaʻawale ʻana, a me ka mālama ʻana i nā poʻe maʻi he mea koʻikoʻi, e koi ana i nā hoʻolālā diagnostic wikiwiki, maʻalahi a pololei [2].ʻO ka hōʻailona o nā maʻi maʻi maʻamau e hui pū i ka hoʻokolohua lapaʻau e pili ana i nā hōʻailona a me nā hōʻailona a me nā noiʻi lab e like me ka moʻomeheu cell a me nā diagnostics molecular, e koi ana i nā limahana i hoʻomaʻamaʻa ʻia, nā kaʻina hana hana, a me nā mea hoʻāʻo kūʻai nui [3, 4].ʻO ka pale ʻana i nā maʻi maʻi maʻi e pono ai ka maʻi maʻi maʻi wikiwiki, maʻalahi, a pololei hoʻi i ka ʻike kūloko, ʻoi aku hoʻi ma nā wahi i kaupalena ʻia i nā maʻi maʻi maʻamau a paʻakikī hoʻi [5], a me ka mālama ʻana i ka wao nahele a i ke kahua kaua, kahi hiki ʻole ke ʻike ʻia nā pilikia..Ua kaupalena ʻia ka mālama olakino [6].I loko o kēia pōʻaiapili, microfluidics kahi ʻenehana e hui pū ai i nā ʻenehana ʻōnaehana microelectromechanical, nanotechnology, a i ʻole nā mea ʻepekema no ka hoʻoheheʻe ʻana i ka wai [7,8,9,10], e hāʻawi ana i nā mea hou no ka ʻike ʻana i ka point-of-care detection (POCT).) nā mea maʻi maʻi ma waho o nā halemai a me nā hale hana.Ke hoʻohālikelike ʻia me nā diagnostics hoʻopau manawa, hāʻawi ka ʻenehana microfluidic i ka laʻana a me ka mālama kālā no nā diagnostics molecular i ka wā o ka maʻi maʻi.ʻO ka hoʻolaha honua ʻana o ka maʻi coronavirus 2019 (COVID-19) ke kumu o ka maʻi maʻi respiratory syndrome coronavirus 2 (SARS-CoV-2), no laila ua hoʻokūpaʻa hou ʻia ke koʻikoʻi o nā microfluidics no ka pale ʻana a me ka mālama ʻana i ka maʻi maʻi maʻi [11, 12] , 13].ʻAʻole like me nā diagnostics kuʻuna, hoʻohana ka microfluidic POCT i nā mea liʻiliʻi liʻiliʻi mai nā mea hoʻopili benchtop a i nā ʻāpana hoʻāʻo ʻaoʻao liʻiliʻi e hoʻāʻo ai ma kahi kokoke i kahi laʻana [14].Hōʻike kēia mau hoʻāʻo i ka hoʻomākaukau ʻana maʻalahi a ʻaʻohe kumu hoʻohālike, ka hoʻonui ʻana i ka hōʻailona wikiwiki, a me nā heluhelu hōʻailona koʻikoʻi e hopena i ka manawa pōkole a me nā hopena pololei i loko o nā minuke.ʻO ka loaʻa ʻana a me ka hana nui ʻana o nā mea mālama ola microfluidic-based i hoʻonui i kā lākou mau kumukūʻai kūpono a pololei i nā noi diagnostic ma waho o ka haukapila, kokoke i ka mea maʻi, a ma ka home.
Ma waena o nā hoʻolālā e kū nei no ka ʻike ʻana i nā maʻi infectious, ʻo ka diagnostics molecular kekahi o nā mea maʻalahi loa [15, 16].Eia kekahi, hoʻohana pinepine ʻia nā diagnostics molecular e like me ke kūlana gula no ka ʻike mau ʻana o COVID-19, e ʻae ana i ka ʻike pololei ʻana i nā wahi kikoʻī o RNA a i ʻole DNA ma mua o ka hoʻomaka ʻana o kahi pane immune [17, 18].Ma ka loiloi o kēia manawa, hōʻike mākou i nā holomua hou loa i nā kaʻina diagnostic molecular microfluidics-based no nā maʻi infectious, mai kahi hiʻohiʻona hoʻonaʻauao i nā hiʻohiʻona ʻoihana e hiki mai ana (Fig. 1).E hoʻomaka mākou me nā ʻanuʻu koʻikoʻi ʻekolu i ka ʻike ʻana i ka waika nucleic: ma ka chip sample pretreatment, nucleic acid amplification, a me ka heluhelu hōʻailona.A laila hoʻohālikelike mākou i nā ʻano like ʻole o ka microfluidic platforms me kā lākou hana a me ka hana, e hōʻike ana i nā hiʻohiʻona kūʻokoʻa (ikaika a me nā nāwaliwali).Kūkākūkā hou ʻia ka ʻike nucleic acid a hāʻawi ʻia ma ke ʻano he kumu hoʻohālike o ke kolu o ka ʻenehana no ka helu piha ʻana o nā molekele pathogen infectious.Eia kekahi, e hōʻike ʻia kekahi mau polokalamu POCT maʻamau a hou loa e hōʻike i ke kūlana o kēia manawa o ka mākeke POCT microfluidic no nā diagnostics molecular.E kūkākūkā pū mākou a wehewehe i kā mākou ʻike no nā noi e hiki mai ana.
Hiki ke hoʻokaʻawale ʻia nā modules o nā microfluidic chips no ka ʻike nucleic acid i ʻekolu mau ʻāpana (sampling, recognition, and signaling) e like me kā lākou hana [19].Ma waena o kēia mau modula, ʻike nui ka module sampling i ka laʻana lysis a me ka ʻokiʻoki nucleic acid.ʻO ka module sensor ka mana nui i ka hoʻololi a me ka hoʻonui ʻana i nā hōʻailona nucleic acid.ʻIke ka module hōʻailona i ka hōʻailona i hoʻololi ʻia a hana ʻia e ka module sensing.Ma muli o ke kaʻina hana o ka ʻike ʻana i nā waikawa nucleic ma kahi chip, e hōʻuluʻulu mākou i nā ʻāpana like ʻole e hiki ke hoʻomaopopo i ka hana "input and output".
ʻO ka ʻanuʻu mua i ka ʻike ʻana i ka waikawa nucleic, ʻo ia ka hoʻokaʻawale ʻana i ka waikawa nucleic i manaʻo ʻia mai ka hāpana kumu.Hana ʻia ka ʻili ʻana o ka waika nucleic e hoʻomaʻemaʻe i nā ʻakika nucleic mai nā mea haumia ʻē aʻe, e hōʻoia i ka paʻa o ka hana mua o nā molekala nucleic acid, a me nā hopena maikaʻi.Pono ka lawe ʻana i ka waika nucleic i ka lysis laʻana e pono ai a me ka hopu ʻana i ka waikawa nucleic, ʻo ka maikaʻi a me ka pono o ia mea he hopena nui i ka noiʻi a me nā hopena diagnostic.Hiki ke kaupalena ʻia nā hopena ʻaoʻao maʻalahi i ka wā o ka unuhi ʻana.ʻO kahi laʻana, ka polymerase chain reaction (PCR) a me ka loop isothermal amplification (LAMP) ua kāohi ʻia e kekahi mau mea hoʻoheheʻe kūlohelohe e like me ka ethanol a me ka isopropanol i loko o ka nucleic acid isolation reagents [20].ʻO ka ʻili wai-wai a me ka hoʻoheheʻe paʻa-paʻa ʻo ia nā ala kaulana loa no ka hoʻokaʻawale ʻana i nā ʻakika nucleic [21], akā naʻe, ʻoi loa ka palena o ka ʻili wai-wai ma kahi chip, no ka mea, ʻo nā reagents i hoʻohana ʻia i ka ʻili wai-wai ke kumu o ka ʻino o ka hapa nui o nā ʻāpana microfluidic. .Maʻaneʻi, hōʻike mākou i nā ʻano hana hoʻokaʻawale paʻa paʻa microarray a hoʻohālikelike i kā lākou pono a me nā hemahema.
ʻO ka silikoni kahi mea substrate kūpono me nā waikawa nucleic ma muli o kona biocompatibility, kūpaʻa, a me ka maʻalahi o ka hoʻololi [22].ʻO ka mea nui, i ka wā i hoʻololi ʻia me ka silica a i ʻole nā mea ʻē aʻe, hōʻike kēia composite i nā waiwai e adsorb negatively charged nucleic acids ma lalo o ka pH haʻahaʻa, nā kūlana paʻakai kiʻekiʻe aʻo ka eluting me ka pH kiʻekiʻe, nā haʻahaʻa paʻakai haʻahaʻa.Ma muli o kēia hanana, hiki ke hoʻomaʻemaʻe i ka waikawa nucleic.
Ua hoʻohana ʻia nā ʻano mea like ʻole o ka silica-based no ka unuhi ʻana o ka nucleic acid i nā microfluidics, e like me nā silica beads, pauka, nā kānana microfiber, a me nā membrane silica [23, 24, 25, 26].Ma muli o nā waiwai o ka mea, hiki ke hoʻohana ʻia nā mea i hoʻokumu ʻia i ka silicon i nā microcircuits i nā ʻano like ʻole.No ka laʻana, hiki ke hoʻokomo wale ʻia nā silica granules, pauka, a me nā nanofilters pāʻoihana i loko o nā pores a i ʻole microchannels o nā microfluidic chips a kōkua i ka unuhi ʻana i nā waikawa nucleic mai nā laʻana [27, 28, 29].Hiki ke hoʻohana ʻia nā membrane silica i hoʻololi ʻia i ka ʻili e hoʻomaʻemaʻe wikiwiki i ka DNA mai nā pathogens ma ke kumu kūʻai haʻahaʻa.No ka laʻana, ʻo Wang et al.[30] Ma ka hoʻohui ʻana i nā hopena hoʻonui denaturing me ka hoʻololi kaulahao-mediated vesicle me nā membrane silica i uhi ʻia me ka chitosan oligosaccharides, ua hoʻokomo ʻia kahi ʻōnaehana lawe lima nui i ʻike maikaʻi ʻia 102-108 colony forming units.(CFU)/ml Vibrio parahaemolyticus., a ua ʻike maʻalahi ka hele ʻana o ka maʻi.Powell et al.[31] Ua hoʻohana ʻia nā microarrays e pili ana i ka silikon no ka ʻike ʻana i ka maʻi hepatitis C (HCV), ka maʻi immunodeficiency kanaka (HIV), ka maʻi maʻi Zika, a me ka papillomavirus kanaka a me ka hoʻolaha maʻalahi, kahi i kūkulu ʻia ai kahi microreactor tortuous 1.3 μl e hopu i nā maʻi RNA.a hana i ka hoʻonui in situ.Ma waho aʻe o kēia mau ʻano, hana nui nā silica microcolumns i hoʻololi ʻia i ka ʻili i ka ʻili o ka nucleic acid, no ka mea, ʻo ka geometry a me nā waiwai o ka mea hoʻololi e hoʻonui nui i ka pono o ka unuhi.ʻO Chen et al.[32] hoʻolālā i kahi microfluidic platform no ka hoʻokaʻawale ʻana i ka RNA haʻahaʻa haʻahaʻa e pili ana i nā microcolumns silicon-coated amino.Hoʻohui kēia mea microfluidic i kahi ʻano o nā micropillars 0.25 cm2 ma kahi substrate silicon e loaʻa ai ka ʻoi aku ka maikaʻi o ka unuhi ʻana ma o kahi kiʻekiʻe kiʻekiʻe i ka hoʻolālā ratio ratio.ʻO ka pōmaikaʻi o kēia hoʻolālā, ʻo ia ka mea hiki i ka microfluidic device ke hoʻokō a hiki i ka 95% nucleic acid extraction efficiency.Hōʻike kēia mau hoʻolālā silika i ka waiwai o ka hoʻokaʻawale wikiwiki ʻana i nā waikawa nucleic ma ke kumu kūʻai haʻahaʻa.I ka hui pū ʻana me nā microfluidic chips, ʻaʻole hiki ke hoʻonui wale i nā hoʻolālā hoʻoheheʻe ʻia e pili ana i ka silicon i ka pono o ka ʻike nucleic acid, akā e hoʻomaʻamaʻa hoʻi i ka miniaturization a me ka hoʻohui ʻana o nā mea analytical [20].
Hoʻohana nā ʻano hoʻokaʻawale magnetic i nā ʻāpana magnetic e hoʻokaʻawale i nā ʻakika nucleic i ke alo o kahi kahua magnetic waho.Hoʻohana pinepine ʻia nā ʻāpana magnetic Fe3O4 a i ʻole γ-Fe2O3 i uhi ʻia me ka silica, amino a me ka carboxyl [33,34,35,36].ʻO ka hiʻohiʻona ʻokoʻa o nā ʻāpana magnetic i hoʻohālikelike ʻia i nā kaʻina SPE e pili ana i ka silicon, ʻo ia ka maʻalahi o ka manipulation a me ka hoʻomalu ʻana me nā magnet o waho.
Ke hoʻohana nei i ka pilina electrostatic ma waena o ka nucleic acids a me ka silica, ma lalo o nā kūlana o ka paʻakai kiʻekiʻe a me ka pH haʻahaʻa, ua hoʻopili ʻia nā waikawa nucleic ma ka ʻili o nā ʻāpana magnetic silica-coated, ʻoiai ma lalo o nā kūlana o ka paʻakai haʻahaʻa a me ka pH kiʻekiʻe, hiki ke holoi ʻia nā molekala. hou..Hiki i nā peʻa magnetic i uhi ʻia i ka silica ke hiki ke unuhi i ka DNA mai nā laʻana leo nui (400 μL) me ka hoʻohana ʻana i ka neʻe magnetically controlled [37].Ma keʻano he hōʻike, Rodriguez-Mateos et al.[38] hoʻohana i nā magnet hiki ke hoʻomalu i ka hoʻoili ʻana o nā peʻa magnetic i nā keʻena like ʻole.Ma muli o ka silica-coated magnetic particles, 470 kope/mL o SARS-CoV-2 genomic RNA hiki ke unuhi ʻia mai nā ʻano wai ʻōpala no ka LAMP reverse transcription detection (RT-LAMP) a hiki ke heluhelu ʻia ka pane i loko o 1 hola.maka olohelohe (Fig. 2a).
Nā mea hana e pili ana i nā mea magnetic a me nā mea porous.ʻO ke kiʻi manaʻo o ka IFAST RT-LAMP microfluidic device no ka ʻike ʻana i ka SARS-CoV-2 RNA (i hoʻololi ʻia mai [38]).b Centrifugal micro device for dSPE of buccal swab nucleic acid (hoʻololi ʻia mai [39]).c Hoʻohana ʻia ka mea hoʻopono ponoʻī i hoʻohana ʻia me kahi kāleka FTA® (i hoʻololi ʻia mai [50]).d Ua hoʻololi ʻia ka pepa kānana Fusion 5 me ka chitosan (i hoʻololi ʻia mai [51]).ʻO SARS-CoV-2 maʻi ʻeha ʻeha koʻikoʻi coronavirus 2, RT-LAMP reverse transcription loop mediated isothermal amplification, FTA finders technology partners, NA nucleic acid
He mea kūpono nā ʻāpana magnetic i hoʻopaʻa ʻia no ka hoʻopili ʻana i ka iwi kuamoʻo phosphate o kahi waikawa nucleic.Ma kahi paʻakai paʻakai, hiki ke hoʻopiʻi maikaʻi ʻia nā pūʻulu phosphate i hoʻopiʻi ʻia o ka nucleic acid ma luna o ka ʻili o nā mea hoʻohui magnetic.No laila, ua hoʻomohala ʻia nā nanoparticles magnetic me kahi ʻili ʻili a me kahi kiʻekiʻe kiʻekiʻe o nā hui amino no ka unuhi ʻana o nā waikawa nucleic.Ma hope o ka hoʻokaʻawale ʻana a me ka hoʻopaʻa ʻana, hiki ke hoʻohana pololei ʻia nā nanoparticles magnetic a me nā DNA complexes i PCR, kahi e hoʻopau ai i ka pono no ka hoʻomaʻemaʻe paʻakikī a me ka hoʻopau manawa a me nā hana elution [35].Ua hoʻohana pū ʻia nā nanoparticles magnetic i uhi ʻia me nā hui carboxyl maikaʻi ʻole e hoʻokaʻawale i nā waikawa nucleic i hoʻopili ʻia ma luna o nā ʻili i ka polyethylene glycol a me ka sodium chloride solutions [36].Me kēia mau peʻa magnetic i hoʻololi ʻia i ka ʻili, kūpono ka unuhi ʻana o DNA me ka hoʻonui ʻana.ʻO Dignan et al.[39] wehewehe ʻia kahi kahua microfluidic centrifugal maʻemaʻe a lawe ʻia no ka nucleic acid pretreatment, e ʻae ana i nā limahana hana ʻole e hoʻohana ia mea ma ka pūnaewele.Eia kekahi, ʻo ka hoʻohālikelike ʻana o ka DNA i hoʻokaʻawale ʻia me LAMP, kahi ala kūpono no ka nānā ʻana i ka wai nucleic acid point-of-care, e hōʻike hou ana i nā pono lako liʻiliʻi a me ke kūpono no nā hoʻokolohua colorimetric (Fig. 2b).
Hāʻawi nā ʻano kuʻuna makeke i ka hiki ke hoʻokaʻawale ʻokoʻa, aia kekahi o ia mau mea i loko o nā mea hoʻoheheʻe waika nucleic automated [KingFisher;ThermoFisher (Waltham, MA, USA), QIAcube® HT;CapitalBio (Beijing, Kina) a me Biomek®;Beckman (Miami, USA).), Florida, USA)].Hiki ke hoʻohana ʻia nā mea maikaʻi o ka hui ʻana i nā peʻa magnetic me nā microfluidics no ka unuhi ʻana i nā ʻakika nucleic, hiki ke holomua i ka hoʻomohala ʻana i nā diagnostics molecular;akā naʻe, ke hilinaʻi nui nei ka hui ʻana o nā peʻa magnetic me nā microfluidics i nā ʻōnaehana hoʻomalu paʻakikī no ka hoʻoponopono pololei ʻana i nā peʻa magnetic, e wehewehe ana i ka kaulana o nā huahana kalepa he nui a pipiʻi, e kaupalena ana i ka hoʻohana hou ʻana o nā peʻa magnetic ma POCT.
Ua hoʻohana pū ʻia kekahi mau mea porous e like me nā kānana nitrocellulose i hoʻololi ʻia, nā kāleka Finders Technology Associates (FTA), nā pepa kānana polyethersulfone, a me nā mea i uhi ʻia i ka glycan no ka ʻike ʻana i ka waikawa nucleic [40, 41, 42, 43, 44].Ua hoʻohana mua ʻia nā mea fibrous porous e like me ka pepa fibrous e hoʻokaʻawale i ka DNA ma o ka hoʻopili kino ʻana i nā molekala DNA lōʻihi me nā fiber.ʻO nā pores liʻiliʻi e alakaʻi i kahi kaohi kino ikaika o nā molekala DNA, e hoʻopilikia maikaʻi i ka hoʻoiho DNA.Ma muli o ka nui o ka pore o ka pepa fibrous, ʻaʻole hiki ke hoʻokō pono i ka pono o ka amplification DNA [45, 46].ʻO ke kāleka FTA he pepa kānana pāʻoihana i hoʻohana ʻia ma ke kahua o ka lāʻau forensic a hoʻohana nui ʻia i nā wahi ʻē aʻe o ka diagnostics molecular.Ma o ka hoʻohana ʻana i ka pepa kānana cellulose i hoʻopili ʻia me nā kemika like ʻole e lyse i nā membrane cell i loko o ka laʻana, pale ʻia ka DNA i hoʻokuʻu ʻia mai ka degradation a hiki i 2 mau makahiki.ʻO ka mea hou loa, ua hoʻomohala ʻia ka pepa cellulose impregnated no ka ʻike ʻana i ka molecular o nā pathogens like ʻole, me SARS-CoV-2, leishmaniasis, a me ka malaria [47,48,49].Hoʻopili pololeiʻia ka HIV i loko o ka plasma i hoʻokaʻawaleʻia, a ua hoʻonuiʻia ka nucleic acid viral i ka FTA® flow membrane i kūkuluʻia i loko o ka concentrator, e hiki ai ke hana pono i ka nucleic acid [50] (Fig. 2c).ʻO ka pilikia nui me ka ʻike ʻana i ka waika nucleic me ka hoʻohana ʻana i nā kāleka FTA, ʻo nā mea kemika e like me guanidine a me isopropanol ke kāohi i nā hopena amplification ma hope.No ka hoʻoponopono ʻana i kēia pilikia, ua kūkulu mākou i ka pepa kānana Fusion 5 chitosan-hoʻololi ʻia, ka mea e hui pū ai i nā pono o ka interlacing kino o nā molekele DNA a me ka pepa kānana fibrous, a me ka electrostatic adsorption o DNA ma nā pūhui chitosan-hoʻololi ʻia e hoʻokō i ka hoʻoneʻe ʻana o ka nucleic acid. ..nā pulupulu kānana [51] (Fig. 2d).Pēlā nō, ʻo Zhu et al.[52] hōʻike i kahi ʻano PCR i hoʻololi ʻia e chitosan e pili ana i kahi ʻōnaehana microfluidic capillary in situ no ka hoʻokaʻawale wikiwiki ʻana a me ka ʻike ʻana i ka RNA maʻi Zika.Hiki ke hoʻopili ʻia nā ʻakika nucleic i loko o kahi lysate i hui ʻia / PCR medium, kēlā me kēia, e pili ana i ka waiwai o ka chitosan.on and off", pane i ka pH.
E like me ka mea i ʻōlelo ʻia ma luna nei, hoʻohui kēia mau hoʻolālā i nā pono o nā ʻano mea paʻa paʻa a hoʻonui i ka pono o ka hoʻoneʻe nucleic acid i nā microfluidics.Ma nā noi kūpono, ʻaʻole pono ka hoʻohana ʻana i kēia mau mea i ka nui, a ʻo ka mālama ʻana i ka ʻili kūpono a i ʻole ka hoʻololi ʻana i ka ʻili o nā mea maʻamau me kēia mau mea hiki ke mālama i kā lākou hana.No laila, ua manaʻo ʻia ʻo ka hoʻokō ʻana i kēia mau hoʻolālā ma hope o kahi haʻawina hoʻokele hiki ke hōʻemi i nā kumukūʻai.
Hoʻohana pinepine ka hoʻāʻo ʻana o ka waikawa nucleic ma nā microfluidic i nā liʻiliʻi liʻiliʻi liʻiliʻi (<100 µl), no laila pono e hoʻonui ʻia nā ʻakika nucleic target me nā ʻimi kikoʻī no ka hoʻololi ʻana i kahi hōʻailona kūpono no ka ʻike ʻana i lalo (optical, uila, a me ka magnetic) [53, 54]. Hoʻohana pinepine ka hoʻāʻo ʻana o ka waikawa nucleic ma nā microfluidic i nā liʻiliʻi liʻiliʻi liʻiliʻi (<100 µl), no laila pono e hoʻonui ʻia nā ʻakika nucleic target me nā ʻimi kikoʻī no ka hoʻololi ʻana i kahi hōʻailona kūpono no ka ʻike ʻana i lalo (optical, uila, a me ka magnetic) [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]. Ke hoʻāʻo nei i nā waikawa nucleic ma nā microfluidic platforms, hoʻohana pinepine ʻia nā helu liʻiliʻi liʻiliʻi (<100 µL), no laila e koi ʻia ka hoʻonui ʻana o nā ʻakika nucleic target me nā probes kūikawā e hoʻololi iā ia i hōʻailona kūpono no ka ʻike ma hope (optical, uila, a me ka magnetic) [53, 54].微流控 平台平台 的 核酸 检测小样本量: <100 μ μ 信号 信号 信号 信号信号 信号 [5] 电学 和和 磁学 信号信号. 54, 电学 54 和 磁学].微流控 平台 上 的 检测 检测 使用 (<100 μ 游 信号 信号探针 信号信号 信号信号 信号, 信号, 54 544 ]. 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]]. ʻO ka ʻike ʻana i nā ʻakika nucleic ma nā platform microfluidic e hoʻohana maʻamau i nā puʻupuʻu liʻiliʻi liʻiliʻi (<100 μl), e koi ana i ka hoʻonui ʻana i nā ʻakika nucleic target me nā probes kūikawā e hoʻohuli iā lākou i mau hōʻailona no ka ʻike ʻana (optical, uila, a me ka magnetic) [53, 54]] .Hiki i ka hoʻonui ʻana o ka waikawa nucleic i nā microfluidics ke wikiwiki i nā hopena, hoʻopaʻa i nā palena ʻike, hōʻemi i nā koi hoʻohālike, a hoʻomaikaʻi i ka pololei ʻike [55, 56].I nā makahiki i hala iho nei, me ka hoʻokō ʻana i ka ʻike wikiwiki a me ka pololei, ua hoʻohana ʻia nā ʻano hana amplification nucleic acid i nā microfluidics, me ka PCR a me kekahi mau hopena isothermal amplification.E hōʻuluʻulu kēia ʻāpana i nā ala no ka ʻike ʻana i ka waikawa nucleic e pili ana i nā ʻōnaehana microfluidic.
ʻO ka PCR kahi hoʻohālikelike o ke kaʻina hana hoʻopiʻi DNA o kahi mea ola, ʻo ke kumumanaʻo i wehewehe ʻia ma nā wahi ʻē aʻe a ʻaʻole e kūkākūkā ʻia ma aneʻi.Hiki i ka PCR ke hoʻonui i kahi helu liʻiliʻi loa o ka DNA/RNA i hoʻopaʻa ʻia ma ka helu exponential, e hana ana i ka PCR i mea hana ikaika no ka ʻike wikiwiki ʻana i nā waikawa nucleic.I nā makahiki i hala iho nei, ua hoʻomohala ʻia nā polokalamu microfluidic portable me nā PCR thermal cycling system e hoʻokō i nā pono o nā diagnostics point-of-care [57, 58].Hiki ke hoʻokaʻawale ʻia ka PCR on-chip i ʻehā mau ʻano (maʻamau, kahe mau, hoʻololi spatially, a me PCR convective) e like me nā ʻano kaohi wela like ʻole [59].No ka laʻana, ʻo Gee et al.[60] hoʻomohala i kahi ʻano PCR (RT-qPCR) hoʻohuli pololei i kā lākou microfluidic platform no ka ʻike nui ʻana o nā maʻi maʻi SARS-CoV-2, influenza A a me B i nā laʻana swab puʻu (Fig. 3a).Park et al.[61] kūkulu ʻia kahi puʻupuʻu loiloi pathogen maʻalahi ma o ka hoʻohui ʻana i ka PCR kiʻiʻoniʻoni lahilahi, nā electrodes, a me kahi module microfluidic-based polydimethylsiloxane i hoʻohana ʻia me ka manamana lima.Eia naʻe, hoʻokomo nā hana ʻelua i nā hemahema maʻamau o PCR maʻamau.Pono ka PCR i ka paikikala wela, kahi e kaupalena ʻia ai ka hoʻonui ʻana o ka mīkini a hoʻemi i ka manawa hoʻāʻo.
He mea koʻikoʻi ka hoʻomohala ʻana o ka microfluidic a me ka PCR hoʻololi hoʻololi hoʻomau e hoʻoponopono i kēia pilikia.Me ka hoʻohana ʻana i kahi kaila serpentine lōʻihi a i ʻole kahi ala pōkole pōkole, hiki i ka PCR kahe mau ke hāʻawi i ka hoʻonui wikiwiki ʻana ma o ka hoʻolalelale ʻana i nā reagents i ʻekolu mau ʻāpana preheat me kahi pauma off-chip.Ke pale pono nei kēia hana i ka pae hoʻololi ma waena o nā mahana hopena like ʻole a pēlā e hoʻemi nui ai i ka manawa hoʻāʻo [62] (Fig. 3b).Ma kekahi haʻawina a Jung et al.[63] hoʻopuka i ka PCR genetic analyser rotary hou e hoʻohui i nā ʻano o ka PCR paʻa a kahe no ka ultrafast a me ka multiplex reverse transcription PCR (Fig. 3c).No ka hoʻonui ʻana i ka waikawa nucleic, e hoʻololi ʻia ka PCR microchip ma o ʻekolu poloka hoʻomehana i nā mahana like ʻole: 1. Palena denaturation 94 ° C, 2. Block Annealing ma 58 ° C, 3. Block expansion ma 72 ° C.
Ka hoʻohana ʻana o PCR i nā microfluidics.Hōʻike schematic o dirRT-qPCR ma kahi microfluidic platform (i hoʻololi ʻia mai [60]).b Hōʻike hoʻohālikelike o kahi microarray PCR kahe mau e pili ana i kahi kahawai serpentine (i hoʻololi ʻia mai [62]).c Hōʻike manaʻo o ka PCR genetic analyzer rotary, ʻo ia hoʻi kahi microchip, ʻekolu poloka hoʻomehana a me kahi kaʻa stepper (i hoʻololi ʻia mai [63]).d Diagram o ka PCR thermoconvection me ka centrifugation a me ka hoʻonohonoho (hoʻololi ʻia mai [64]).ʻO DirRT-qPCR, ka hoʻihoʻi ʻana i ke kaulahao polymerase chain reaction
Me ka hoʻohana ʻana i nā capillaries a me nā puka lou a i ʻole nā papa lahilahi, hiki i ka convection PCR ke hoʻonui wikiwiki i nā ʻakika nucleic e ka convection thermal manuahi kūlohelohe me ka ʻole o ka pono o kahi pamu waho.No ka laʻana, ua hoʻomohala ʻia kahi ʻōnaehana microfluidic cyclic olefin polymer microfluidic ma luna o kahi kahua hoʻomehana hoʻololi i hana ʻia e hoʻohana ana i ka paikikala wela me ka centrifugation i loko o kahi PCR loop microchannel [64] (Fig. 3d).Hoʻoulu ʻia ka hopena hopena e ka convection wela, e hoʻololi mau ana i ka wela kiʻekiʻe a me ka haʻahaʻa i loko o kahi microchannel me kahi ʻano annular.Hiki ke hoʻopau ʻia ke kaʻina hana amplification holoʻokoʻa i 10 mau minuke me ka palena ʻike o 70.5 pg/channel.
E like me ka mea i manaʻo ʻia, ʻo ka PCR wikiwiki kahi mea hana ikaika no ka hoʻopili piha ʻana i ka sample-pane molecular diagnostic and multiplex analysis system.Hoʻemi nui ka Rapid PCR i ka manawa e pono ai e ʻike iā SARS-CoV-2, ka mea e hāʻawi i ka hoʻomalu pono o ka maʻi maʻi COVID-19.
Pono ka PCR i kahi kaʻa kaʻa wela paʻakikī i kūpono ʻole no POCT.ʻO ka mea hou aku nei, ua hoʻohana ʻia nā ʻenehana hoʻonui isothermal i nā microfluidics, e komo pū ana akā ʻaʻole i kaupalena ʻia i ka LAMP, recombinase polymerase amplification (RPA), a me ka amplification e pili ana i ka nucleic acid sequences [65,66,67,68].Me kēia mau ʻenehana, hoʻonui ʻia nā nucleic acid i kahi mahana mau, e hoʻomaʻamaʻa i ka hana ʻana i nā mea haʻahaʻa haʻahaʻa, nā hāmeʻa POCT portable koʻikoʻi no nā diagnostics molecular.
Hiki i nā hōʻike LAMP e pili ana i ka microfluidics kiʻekiʻe ke ʻike i nā maʻi maʻi maʻi [42, 69, 70, 71].I ka hui pū ʻana me kahi ʻōnaehana microfluidic centrifugal, hiki i ka LAMP ke hoʻomaʻamaʻa hou i ka automation o ka ʻike wai nucleic [69, 72, 73, 74, 75].Ua hoʻomohala ʻia ka spin-and-react SlipChip no ka ʻike maka ʻana o nā hua bacteria parallel lehulehu me ka hoʻohana ʻana i ka LAMP [76] (Fig. 4a).I ka hoʻohana ʻana i ka LAMP i hoʻopaʻa ʻia i ka assay, ʻo ka fluorescence signal-to-noise ratio ma kahi o 5-fold, a hiki i ka palena ʻike i ka 7.2 kope / μl o genomic DNA. Eia kekahi, ʻo ka noho ʻana o ʻelima mau maʻi bacterial digestive maʻamau, me Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis a me Vibrio parahaemolyticus, ua ʻike ʻia ma muli o ke ʻano ma <60 min. Eia kekahi, ʻo ka noho ʻana o ʻelima mau maʻi bacterial digestive maʻamau, me Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis a me Vibrio parahaemolyticus, ua ʻike ʻia ma muli o ke ʻano ma <60 min.Eia kekahi, ʻo ka hele ʻana o ʻelima mau maʻi bacterial maʻamau o ka digestive tract, me Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis a me Vibrio parahaemolyticus, ua ʻike ʻia me ka hoʻohana ʻana i kēia ʻano ma lalo o 60 mau minuke.此外, 基于 该 方法 在 <24 分钟 可 视化 细菌病 细菌病消化道 的 存在存在, 弧菌 河流 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌.此外, 基于 该 方法 在 <24 分钟 视化 了 消化道 消化道 存在细菌病, 包括 氏 弧菌副溶血副溶血 性 ... 弧菌 弧菌 弧菌弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 HIPEia kekahi, ua ʻike ʻia ka loaʻa ʻana o ʻelima mau maʻi maʻi bacterial maʻamau, me Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvius, a me Vibrio parahaemolyticus, i ʻike ʻia me ka hoʻohana ʻana i kēia ʻano ma lalo o 60 mau minuke.
ʻO nā mea maikaʻi o ka LAMP i nā microfluidics, me nā mea ʻē aʻe, ka pane wikiwiki a me ka ʻike miniaturized.Eia nō naʻe, ma muli o ka wela o ka hopena (ma kahi o 70 ° C), hiki ke hana ʻia nā aerosol i ka wā LAMP, e loaʻa ana i kahi helu kuhi hewa kiʻekiʻe.Pono e hoʻonui ʻia ka ʻike kikoʻī, ka hoʻolālā mua, a me ka mālama ʻana i ka mahana no LAMP.Eia kekahi, he waiwai nui nā hoʻolālā chip e hoʻokō ana i ka ʻike maka nui ma ka pahu hoʻokahi a pono e hoʻomohala ʻia.Eia kekahi, kūpono ʻo LAMP no ka ʻike ʻana i nā kumu he nui i hoʻohui ʻia i hoʻokahi chip, he mea koʻikoʻi nui ia, akā aia nō ka nui o ka lumi no ka hoʻomohala ʻana.
Hiki ke hoʻemi ʻia ka nui o ka maikaʻi wahaheʻe o LAMP me RPA, no ka mea, ʻo ka hopena haʻahaʻa haʻahaʻa haʻahaʻa (~ 37 °C) ka hopena i nā pilikia evaporation liʻiliʻi [77].I loko o ka ʻōnaehana RPA, ʻelua mau kumu mua e hoʻomaka i ka synthesis DNA ma ka hoʻopaʻa ʻana i kahi recombinase a hiki ke hoʻopau ʻia i loko o 10 mau minuke [78,79,80,81].No laila, ʻoi aku ka wikiwiki o ka hana RPA holoʻokoʻa ma mua o PCR a i ʻole LAMP.I nā makahiki i hala iho nei, ua hōʻike ʻia ka ʻenehana microfluidic e hoʻomaikaʻi hou i ka wikiwiki a me ka pololei o RPA [82,83,84].No ka laʻana, Liu et al.[85] hoʻomohala i kahi microfluidic integrated lateral flow polymerase recombinase amplification assay no ka ʻike wikiwiki a paʻakikī hoʻi o SARS-CoV-2 ma o ka hoʻohui ʻana i ka reverse transcription RPA (RT-RPA) a me kahi ʻōnaehana hōʻike hōʻike hōʻike ākea ākea ākea.i loko o kahi ʻōnaehana microfluidic hoʻokahi.Helu 4b).ʻO ka palena o ka ʻike ʻana he 1 kope/µl a i ʻole 30 kope/lapana, a hiki ke hoʻopau ʻia ka ʻike ma kahi o 30 mau minuke.Kong et al.ua hoʻomohala i kahi mea microfluidic hiki ke komo.[86] hoʻohana i ka wela o ke kino a me kahi ʻōnaehana ʻike fluorescence e pili ana i ke kelepona paʻa e ʻike wikiwiki a ʻike pololei i ka HIV-1 DNA me ka hoʻohana ʻana i ka RPA (Figure 4c).ʻIke ʻia ka hoʻāʻo RPA hiki ke hoʻohana ʻia he 100 kope/mL o ke kaʻina i hoʻopaʻa ʻia i loko o 24 mau minuke, e hōʻike ana i ka mana nui no ka hoʻomaʻamaʻa wikiwiki ʻana i nā pēpē HIV-1-i maʻi i nā ʻōnaehana palena waiwai.
ʻO ka hoʻonui ʻana i ka isothermal i ka hoʻāʻo ʻana i ka point-of-care testing (POCT).Ka hoʻomohala ʻana a me ka hana ʻana o ka spin a me ka hopena SlipChip.Ma hope o ka hoʻoheheʻe ʻana i ka plasma, ua hui pū ʻia nā ʻāpana o luna a me lalo me kahi pūʻulu o nā nati e hana i ka chip hope loa (i hoʻololi ʻia mai [76]).b Schematic o ka ʻōnaehana MI-IF-RPA no ka ʻike COVID-19 (i hoʻololi ʻia mai [85]).c Hoʻolālā o kahi hōʻike RPA hiki ke hoʻohana ʻia no ka ʻike wikiwiki ʻana i ka HIV-1 DNA (i hoʻololi ʻia mai [86]).SE Salmonella enterica, VF Vibrio fluvius, VP Vibrio parahaemolyticus, BC Bacillus cereus, EC Escherichia coli, FAM carboxyfluorescein, virus immunodeficiency kanaka HIV, RPA recombinase polymerase amplification, LED light emitting diode, MI-IF-RPA Microfluidics F Integrated Lacombina Hoʻohuihui. Hoʻonui
Ke ulu wikiwiki nei ka RPA e pili ana i ka microfluidic, akā naʻe, ʻoi aku ka kiʻekiʻe o ke kumukūʻai o ka hana ʻana i ka chip a me ka hoʻohana ʻana i ka hopena a pono e hoʻemi ʻia e hoʻonui i ka loaʻa o kēia ʻenehana.Eia kekahi, hiki i ke kiʻekiʻe kiʻekiʻe o RPA ke hoʻopili i ka hoʻonui ʻana i nā huahana kikoʻī ʻole, ʻoi aku hoʻi i ke alo o ka contamination.Hiki i kēia mau palena ke hoʻopili i ka hoʻohana ʻana o RPA i nā ʻōnaehana microfluidic a pono e hoʻonui hou ʻia.Pono nō hoʻi nā primers i hoʻolālā maikaʻi ʻia no nā pahuhopu like ʻole e hoʻomaikaʻi i ka hiki ke kūpono o nā hoʻolālā microfluidic e pili ana i ka RPA ma POCT.
Loaʻa iā Cas13 a me Cas12a ka hiki ke hoʻokaʻawale i nā ʻakika nucleic a no laila hiki ke hoʻomohala ʻia ma ke ʻano he ʻike a me nā mea hana diagnostic.Hoʻohana ʻia ʻo Cas13 a me Cas12a i ka hoʻopaʻa ʻana i ka DNA a i ʻole RNA.I ka manawa e ho'ā ai, hoʻomaka ka pūmua e hoʻokaʻawale i nā ʻakika nucleic ʻē aʻe e pili ana, a ma hope o ke alakaʻi ʻana i nā RNA e kuhikuhi ana i nā ʻakika nucleic pathogen-specific hiki ke hoʻokaʻawale i nā probes fluorescent pio a hoʻokuʻu i ka fluorescence.Ma muli o kēia kumumanaʻo, Kellner et al.[87] hoʻomohala i kahi ʻano hana Cas13 [Specific High-sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK)], a me Broughton et al.[88] hoʻomohala i kahi ala ʻē aʻe e pili ana i Cas12a [CRISPR Trans Reporter targeting DNA endonuclease (DTECR)].
I nā makahiki i hala iho nei, ua ʻike ʻia nā ʻano hana like ʻole no ka ʻike ʻana i nā waikawa nucleic e pili ana i ka CRISPR [89, 90].ʻO nā hana maʻamau CRISPR e hoʻohana pinepine ana i ka manawa a me ka hana ikaika ma muli o nā kaʻina hana he nui e pili ana i ka ʻili nucleic acid, amplification a me ka ʻike CRISPR.Hiki ke hoʻonui ʻia ka loaʻa ʻana o nā hopena maikaʻi ʻole i ka ʻike ʻana o ka wai i ka ea.Hāʻawi ʻia i ka mea i luna, pono nui nā ʻōnaehana CRISPR i ka optimization.
Ua hoʻomohala ʻia kahi kahua microfluidic pneumatically controlled e hiki ke hana i nā kānana 24 i ka like no nā noi ʻike CRISPR-Cas12a a me CRISPR-Cas13a [91].Hoʻolako ʻia ka ʻōnaehana me kahi mea ʻike fluorescence e kāpae ana i ka hoʻonui ʻana i ka nucleic acid a ʻike maʻalahi i ka femtomolar DNA a me RNA samples.ʻO Chen et al.[92] hoʻohui hou i ka hoʻonui ʻana me ka ʻōnaehana CRISPR-Cas12a i nā microfluidics centrifugal (Fig. 5a).Ua lanakila kēia hana i ka paʻakikī o ka hoʻohui ʻana i kēia mau kaʻina hana ʻelua no ka mea hiki iā Cas12a ke ʻeli i ka DNA ʻelele a keʻakeʻa i ke kaʻina hana amplification.Eia kekahi, ʻo Chen et al.[92] hoʻopaʻa mua i nā mea hoʻonā i loko o kahi mana microfluidic centrifugal e hoʻopau pono i ke kaʻina holoʻokoʻa.Ma kekahi hana, Silva et al.[93] hoʻomohala i kahi ala diagnostic me ka hoʻonui ʻole ʻana o CRISPR/Cas12a a me kahi atamai e ʻike ai iā SARS-CoV-2 (Fig. 5b).ʻO kēia ho'āʻo, i ʻike ʻia he ʻōnaehana hoʻonui ʻole ma ke kelepona, aia kahi enzyme CRISPR/Cas-dependent i hoʻokumu ʻia ma ka ʻike maka ʻana o nā hōʻailona bubble i hana ʻia e catalase i nā kahawai microfluidic.ʻO ka ʻike makaʻala ʻaʻole i emi iho ma mua o 50 kope/µl o ka nucleic acid me ka hoʻonui ʻole ʻana, ʻo ke kaʻina holoʻokoʻa mai ka hāpana ʻana a hiki i ka heluhelu hōʻailona he 71 mau minuke wale nō.
Nā ʻano hana ʻike waika nucleic ma muli o CRISPR.Centrifugal POCT no nā diagnostics molecular integrated e pili ana i ka CRISPR (i hoʻololi ʻia mai [92]).b Ka hoʻomohala ʻana i ka hoʻāʻo CASCADE no ka nānā ʻana i ka polokalamu kelepona o SARS-CoV-2 (i hoʻololi ʻia mai [93]).Hoʻonui hou ʻia ka RAA recombinase, PAM pili protospacer kumu, CRISPR puʻupuʻu palindromic pōkole i nā manawa maʻamau, ʻōnaehana CASCADE me ka hoʻonui ʻole kelepona kelepona me nā enzymes hilinaʻi CRISPR/CAS, 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride EDC
E like me ka hana hope loa i ka ʻike ʻana i ka waikawa nucleic, hōʻike pololei ka ʻike hōʻailona i nā hopena diagnostic a he mea koʻikoʻi ia i ka hoʻomohala ʻana i kahi POCT kūpono, paʻakikī, a pololei.Hiki ke heluhelu ʻia nā hōʻailona me ka hoʻohana ʻana i nā ʻano like ʻole e like me ka fluorescent, electrochemical, colorimetric a me nā hoʻolālā magnetic.Ma kēia ʻāpana, wehewehe mākou i ke kumu no kēlā me kēia ala a hoʻohālikelike i nā diagnostics molecular o nā maʻi infectious i microfluidics.
Hoʻohana nui ʻia nā hoʻolālā e pili ana i ka fluorescence no ka POCT diagnostics o nā maʻi infectious ma muli o ko lākou mau pōmaikaʻi maikaʻi loa o ka naʻau maikaʻi, ka haʻahaʻa haʻahaʻa, ka maʻalahi o ka hana, a me ka nānā ʻana o ka mālama ʻana [94, 95].Hoʻohana kēia mau hoʻolālā i nā fluorophores i hōʻailona ʻia e like me nā mea kala fluorescent a me nā nanomaterials e hana i kahi hōʻailona hiki ke ʻike ʻia (hoʻonui a hoʻopau paha i ka fluorescence).Hōʻike kēia ʻike e hiki ke hoʻokaʻawale ʻia nā hoʻolālā e pili ana i ka fluorescence i ka lepili fluorescent pololei, hōʻailona-i, a me ka ʻike ʻana i ka fluorescent hōʻailona [96].Hoʻohana ka ʻike ʻana i ka lepili fluorescent i nā lepili fluorescent kūikawā e lepili i nā ligand kikoʻī e hoʻohua i ka nui o ka fluorescence ke hoʻopaʻa ʻia i kahi pahu hopu.No ka ʻike ʻana i ka fluorescence e pili ana i ka hōʻailona, pili maikaʻi ka maikaʻi o ka hōʻailona fluorescent i ka nui o ka hoihoi.ʻAʻole liʻiliʻi ka ikaika o ka fluorescence i ka loaʻa ʻole o kahi pahu hopu a ʻike ʻia ke ʻike ʻia ke loaʻa ka nui o ka pahuhopu.ʻO ka ʻokoʻa, ʻo ka ikaika o ka fluorescence i ʻike ʻia e ka "signal-off" fluorescence, ua like ia me ka nui o ka pahu hopu, i ka hoʻomaka ʻana i ka waiwai nui a me ka emi iki ʻana i ka wā e hoʻonui ʻia ai ka pahu hopu.No ka laʻana, me ka hoʻohana ʻana i ka CRISPR-Cas13a target-dependent trans-cleavage mechanism, Tian et al.[97] hoʻomohala i kahi hoʻolālā ʻike hou no ka ʻike ʻana i nā RNA e kāʻalo pololei i ka transcription hoʻohuli (Fig. 6a).Ma ka hoʻopaʻa ʻana i nā RNA i hoʻokō ʻia, hiki ke hoʻāla ʻia ka paʻakikī CRISPR-Cas13-RNA, e hoʻoulu ai i ka cleavage transcollateral e nā RNA mea hoʻolaha ʻole kikoʻī.Hoʻopiʻi ʻia ka mea hoʻolaha i kapa ʻia ʻo fluorescently [fluorophore (F)] e ka quencher (Q) i paʻa a lele i ka wā i hoʻokaʻawale ʻia e ka paʻakikī i hoʻāla ʻia.
ʻO ka pōmaikaʻi o ka ʻike electrochemical ka wikiwiki ʻike kiʻekiʻe, hana maʻalahi, haʻahaʻa kumu kūʻai, maʻalahi ke lawe a me ka mana maʻalahi.ʻO ia ke ʻano loiloi ikaika no nā noi POCT.Ma muli o ka graphene field-effect transistors Gao et al.[98] hoʻomohala i kahi nanobiosensor no ka ʻike nui ʻana o nā antigens maʻi Lyme mai ka bacteria Borrelia burgdorferi me ka palena ʻike o 2 pg / mL (Fig. 6b).
Ua hoʻohana ʻia nā hōʻike colorimetric i nā noi POCT, e pōmaikaʻi ana i nā pono o ka portability, ke kumu kūʻai haʻahaʻa, ka maʻalahi o ka hoʻomākaukau, a me ka heluhelu ʻike.Hiki i ka ʻike kala ke hoʻohana i ka oxidation o peroxidase a i ʻole peroxidase-like nanomaterials, ka hōʻuluʻulu ʻana o nā nanomaterials, a me ka hoʻohui ʻana o nā dyes hōʻailona e hoʻololi i ka ʻike e pili ana i ka loaʻa ʻana o nā waikawa nucleic i ʻike ʻia i nā loli kala [99, 100, 101].ʻO ka mea nui, hoʻohana nui ʻia nā nanoparticles gula i ka hoʻomohala ʻana i nā hoʻolālā colorimetric, a ma muli o ko lākou hiki ke hoʻoulu i ka loli wikiwiki a me ka nui o ka waihoʻoluʻu, ke piʻi nei ka hoihoi i ka hoʻomohala ʻana i nā kahua kala POCT no ka maʻi maʻi infectious [102].Me kahi mea hana microfluidic centrifugal i hoʻohui ʻia [103], hiki ke ʻike ʻia nā pathogens i lawe ʻia i ka meaʻai i loko o nā laʻana waiū i hoʻohaumia ʻia ma ke kiʻekiʻe o nā cell bacterial 10, a hiki ke heluhelu ʻia nā hopena i loko o nā minuke 65 (Fig. 6c).
Hiki i nā ʻenehana hoʻonaʻauao magnet ke ʻike pololei i nā analyte me ka hoʻohana ʻana i nā mea magnetic, a ua nui ka hoihoi i nā noi POCT i nā makahiki i hala iho nei.Loaʻa i nā ʻenehana ʻike magnetic kekahi mau pono kūʻokoʻa e like me ke kumu kūʻai haʻahaʻa magnetic ma mua o nā ʻāpana optical pipiʻi.Eia nō naʻe, ʻo ka hoʻohana ʻana i kahi māla magnetic e hoʻomaikaʻi i ka pono ʻike a hōʻemi i ka manawa hoʻomākaukau laʻana [104].Eia kekahi, ʻo nā hopena o ka hoʻāʻo magnetic e hōʻike i ka kikoʻī kiʻekiʻe, ka naʻau, a me ka lākiō hōʻailona-a-noise kiʻekiʻe ma muli o ka hōʻailona ʻike ʻole magnetic o nā ʻano biological [105].Sharma et al.ua hoʻohui pū i kahi biosensor e pili ana i ka biosensor i loko o kahi paepae microchip portable.[106] no ka ʻike nui ʻana o nā pathogens (Fig. 6d).ʻIke ʻia nā biosensors i nā ʻakika nucleic subnanomolar i hoʻokaʻawale ʻia mai nā pathogens.
ʻAno ʻike hōʻailona maʻamau.ʻO ka manaʻo o ka ʻike hyperlocalized o Cas13a (hoʻololi ʻia mai [97]).b Graphene nanobiosensor FET i hui pū me Lyme GroES scFv (hoʻololi ʻia mai [98]).c Nā hōʻailona colorimetric no ka ʻike nui ʻana o nā pathogens lawe ʻia i ka meaʻai i loko o kahi microfluidic chip centrifugal: No. 1 a me No. 3 samples me nā pathogens target, a me No. 2, No. 4 a me No. 5 nā laʻana me ka ʻole o nā pathogens target (hoʻololi ʻia mai [103]) .d Biosensor e pili ana i kahi hui ʻana o ka tunnel magnetic, me kahi paepae, kahi mea hoʻokiʻekiʻe paʻa i kūkulu ʻia, kahi ʻāpana hoʻomalu, a me kahi lako mana no ka hoʻokumu ʻana i nā hōʻailona (i hoʻololi ʻia mai [106]).GFET Graphene FET, Escherichia coli, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus, Vibrio parahaemolyticus, Listeria monocytogenes, PC PC, PDMS Dimethicone, PMMA polymethyl methacrylate
ʻOiai nā hiʻohiʻona maikaʻi loa o nā ʻano ʻike ma luna, aia nō nā hemahema.Hoʻohālikelike ʻia kēia mau ʻano (papa 1), me kekahi mau noi me nā kikoʻī (pono a maikaʻi ʻole).
Me ka hoʻomohala ʻana o nā microfluidics, nā ʻōnaehana microelectromechanical, nanotechnology a me nā ʻepekema waiwai, ke hoʻohana mau nei ka hoʻohana ʻana i nā ʻāpana microfluidic no ka ʻike ʻana i nā maʻi maʻi maʻi e holomua mau ana [55,96,107,108].ʻO ka hoʻoponopono pololei ʻana i nā mea hana liʻiliʻi a me nā wai e hāʻawi i ka pololei diagnostic a me ka uku-pono.No laila, no ka hoʻomohala hou ʻana, ua hoʻoikaika ʻia e hoʻomaikaʻi a hoʻomaikaʻi i nā chips, ka hopena i nā ʻāpana microfluidic like ʻole me nā hale a me nā hana like ʻole.Ma ʻaneʻi mākou e hoʻolauna pōkole i kekahi mau ʻano maʻamau o nā platform microfluidic a hoʻohālikelike i ko lākou mau hiʻohiʻona (pono a me nā mea ʻino).Eia kekahi, ʻo ka hapa nui o nā hiʻohiʻona i helu ʻia ma lalo nei e pili ana i ka hakakā ʻana iā SARS-CoV-2.
ʻO nā LOCC nā ʻōnaehana analytical paʻakikī liʻiliʻi maʻamau a ʻoi aku ka liʻiliʻi o kā lākou hana, hoʻohui ʻia, automated a hoʻohālikelike ʻia mai ka hoʻohālikelike ʻana a me ka hoʻomākaukau ʻana, ka mana kahe a me ka ʻike wai [109, 110].Hoʻohana ʻia nā wai wai ma o ka geometry i hoʻolālā ʻia a me ka launa pū ʻana o nā hopena kino e like me nā gradients pressure, capillary action, electrodynamics, magnetic fields and acoustic waves [111].Hōʻike ka LOCC i nā pōmaikaʻi maikaʻi loa i ka kānana kiʻekiʻe a me ka ʻike he nui, me ka wikiwiki o ka nānā ʻana, ka liʻiliʻi liʻiliʻi, ka hoʻohana haʻahaʻa haʻahaʻa, a me ka hoʻokele kiʻekiʻe a me ka maikaʻi o ka hana;akā naʻe, maʻalahi loa nā mea LOCC, a me ka hana ʻana, ka paʻi ʻana, a me ka hoʻopili ʻana.Eia nō naʻe, ʻo ka multiplexing a me ka hoʻohana hou ʻana i nā pilikia nui [96].Ke hoʻohālikelike ʻia me nā paepae ʻē aʻe, loaʻa i ka LOCC nā pono kūʻokoʻa e pili ana i ka ʻokoʻa o ka noi a me ka hoʻohālikelike ʻenehana maikaʻi loa, akā ʻike ʻia kona mau hemahema, ʻo ia hoʻi ka paʻakikī kiʻekiʻe a me ka hoʻihoʻi maikaʻi ʻole.ʻO ka hilinaʻi ʻana i nā pamu o waho, ʻoi aku ka nui a me ke kumukūʻai, e kaupalena hou i kā lākou hoʻohana ʻana ma POCT.
I ka wā o ka maʻi maʻi COVID-19, loaʻa ka LOCC i ka nānā nui.I ka manawa like, aia kekahi mau chips hou e hoʻohui i nā ʻenehana.No ka laʻana, ua hoʻohana nui ʻia nā smartphones e like me nā polokalamu analytics portable a loaʻa iā lākou ka mana nui no ka hoʻohui LOCC.ʻO Sun et al.[21] hana i kahi puʻupuʻu microfluidic e hiki ai i ka multiplexing kikoʻī nucleic acid sequences o ʻelima pathogens, me SARS-CoV-2, me ka hoʻohana ʻana i ka LAMP a nānā iā lākou me ka hoʻohana ʻana i kahi kelepona i loko o 1 hola ma hope o ka hopena o ka hopena.E like me kekahi laʻana, Sundah et al.[112] hana i ka molecular switch [catalytic amplification by molecular transition state switch (CATCH)] no ka ʻike pololei a me ka maʻalahi o nā pahuhopu SARS-CoV-2 RNA me ka hoʻohana ʻana i nā kelepona. Hoʻopili ʻia ʻo CATCH me LOCC portable a loaʻa i ka hana ʻoi aku ka maikaʻi (ma kahi o 8 RNA kope/μl; <1 h ma ka lumi wela) [112]. Hoʻopili ʻia ʻo CATCH me LOCC portable a loaʻa i ka hana ʻoi aku ka maikaʻi (ma kahi o 8 RNA kope/μl; <1 h ma ka lumi wela) [112]. CATCH совместим с портативным LOCC и обеспечивает превосходную производительность (примерно 8 копий РНК/мклт; Hoʻopili ʻia ʻo CATCH me LOCC portable a hāʻawi i ka throughput maikaʻi loa (ma kahi o 8 RNA kope/µl; <1 h ma ka lumi wela) [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/мкпл; < 1 темтре ). Ua kūpono ʻo CATCH me nā LOCC portable a he hana maikaʻi loa kona (ma kahi o 8 RNA kope/µl; <1 hola ma ka lumi wela) [112].Eia kekahi, hoʻohana pū nā mea LOCC no nā diagnostics molecular i kekahi mau mea hoʻokele e like me ka vacuum, stretch, a me nā māla uila.Kang et al.[113] hōʻike i kahi PCR nanoplasma-on-a-chip i ka manawa maoli no ka maʻi wikiwiki a me ka nui o ka COVID-19 ma ke kula me ka hoʻohana ʻana i kahi pahu PCR wai plasmonic vacuum.Li et al.[114] a laila hoʻomohala i kahi puʻupuʻu microfluidic i hoʻopaʻa ʻia e hiki ai i ka maʻi o COVID-19.Hoʻohana ka paepae i ka ʻōnaehana hoʻonui RT-LAMP e hoʻoholo ai inā maikaʻi a maikaʻi ʻole paha kahi laʻana.Ma hope mai, ʻo Ramachandran et al.[115] loaʻa i nā gradients kahua uila kūpono e hoʻohana ana i ka isotachophoresis (ITP), kahi ʻenehana koho ion i hoʻokō ʻia i nā microfluidics.Me ka ITP, hiki ke ho'oma'ema'e 'akomi 'ia ka RNA o ka nasopharyngeal swab.A laila ʻo Ramachandran et al.[115] ʻO ka hui pū ʻana i kēia hoʻomaʻemaʻe ITP me ITP-enhanced LAMP a me CRISPR assays i ʻike ʻia ʻo SARS-CoV-2 i loko o ka swab nasopharyngeal kanaka a me nā specimens clinical ma kahi o 35 mau minuke.Eia kekahi, ke puka mau nei nā manaʻo hou.ʻO Jadhav et al.[116] hoʻolālā i kahi hoʻolālā diagnostic e pili ana i ka spectroscopy Raman i hoʻonui ʻia i ka ʻili i hui pū ʻia me kahi mea microfluidic i loaʻa i nā nanotubes kalapona i hoʻopaʻa ʻia i ke gula a i ʻole nā notubes electrospun hiki ke hoʻohana.Hiki ke hoʻopau ʻia nā microchannels kānana i hana ʻia me ka membrane.Hoʻopili ka hāmeʻa i nā maʻi mai nā wai kino like ʻole e like me ka saliva, nasopharynx a me nā waimaka.No laila, ʻoi aku ka nui o ka titer virus a hiki ke ʻike pololei ʻia ka maʻi ma ka pūlima Raman.
ʻO LOAD kahi kahua microfluidic centrifugal kahi e mālama ʻia ai nā kaʻina āpau e kahi protocol frequency e hoʻololi i kahi substrate microstructured [110].Hōʻike ʻia ka mea LOAD me ka hoʻohana ʻana i ka ikaika centrifugal ma ke ʻano he mea hoʻokele koʻikoʻi.Hoʻopili ʻia nā wai i nā kaha capillary, Euler a me Coriolis.Ke hoʻohana nei i kahi mea centrifuge, hana ʻia nā loiloi i ka hana hoʻomau wai mai kahi radial i loko a i waho kūlana, e hoʻopau ana i ka pono o nā paipu waho waho, nā pamu, nā mea hana, a me nā kiwi hana.I ka pōkole, hoʻokahi ala hoʻomalu e maʻalahi i ka hana.ʻO nā ikaika e hana ana ma luna o ka wai i loko o ke kahawai microfluidic like ma ka mamao like mai ke kikowaena haʻahaʻa ua like, e hiki ai ke hana hou i ke ʻano o ke kahawai.No laila, ʻoi aku ka maʻalahi o ka lako LOAD a ʻoi aku ka maikaʻi o ka hoʻolālā ʻana a me ka hana ʻana ma mua o nā lako LOCC maʻamau, ʻoiai ka nui o nā ʻano kūʻokoʻa a hoʻohālikelike ʻia;akā naʻe, ma muli o ka ikaika mechanical kiʻekiʻe o nā mea centrifugal, ua kaupalena ʻia nā mea chip i loaʻa a paʻakikī nā puke liʻiliʻi.i ke kaʻa.Ma ka manawa like, ua hoʻolālā ʻia ka hapa nui o nā mea LOAD no ka hoʻohana hoʻokahi wale nō, he kumukūʻai no ka ʻike nui [96, 117, 118, 119].
I nā makahiki i hala iho nei, LOAD, i manaʻo ʻia kekahi o nā mea hana microfluidic maikaʻi loa, ua loaʻa i ka nānā nui ʻana mai nā mea noiʻi a me nā mea hana.No laila, ua loaʻa ka LOAD i ka ʻae ākea a ua hoʻohana ʻia no ka diagnostics molecular o nā pathogens infectious [120, 121, 122, 123, 124], ʻoi aku hoʻi i ka wā o ka hanana COVID-19.No ka laʻana, i ka hopena o 2020, Ji et al.Ua hōʻike ʻo [60] i kahi hōʻike RT-qPCR pololei no ka ʻike wikiwiki a me ka automated parallel detection o SARS-CoV-2 a me ka maʻi influenza A a me B i nā ʻāʻī swab specimens.A laila ʻo Xiong et al.[74] hōʻike i kahi LAMP-integrated discoid microfluidic platform no ka wikiwiki, pololei, a me ka manawa like o nā coronaviruses hanu kanaka ʻehiku, me SARS-CoV-2, i loko o 40 mau minuke.I ka hoʻomaka ʻana o 2021, de Oliveira et al.[73] hōʻike i ka polystyrene toner centrifugal microfluidic chip, hana lima ʻia me ka manamana lima, no ka RT-LAMP molecular diagnostic o COVID-19.Ma hope mai, ʻo Dignan et al.[39] hōʻike i kahi microdevice centrifuge portable automated no ka hoʻomaʻemaʻe ʻana i ka SARS-CoV-2 RNA pololei mai nā ʻāpana buccal swab.ʻO Medved et al.[53] hoʻolālā i kahi ʻōnaehana sampling aerosol SARS-CoV-2 inline me kahi puʻupuʻu liʻiliʻi e hoʻololi ana i ka microfluidic fluorescent chip me ka palena ʻike o 10 kope / μL a me ka paepae pōkole liʻiliʻi o 15 mau minuke.Suarez et al.[75] Ua hōʻike hou aku nei i ka hoʻomohala ʻana o kahi papa hana microfluidic modular modular no ka ʻike pololei ʻana o SARS-CoV-2 RNA i nā laʻana nasopharyngeal swab wela-inactivated me ka hoʻohana ʻana i ka LAMP.Hōʻike kēia mau hiʻohiʻona i nā pōmaikaʻi nui a me ka ʻōlelo hoʻohiki o LOAD i ka diagnostics molecular o COVID-19.
I ka makahiki 1945, ua hōʻike mua ʻo Muller lāua ʻo Clegg [125] i nā kahawai microfluidic ma ka pepa me ka pepa kānana a me ka paraffin.Ma 2007, ua hana ka hui Whitesides [126] i ka paepae pepa hana mua no ka ho'āʻo ʻana i ka protein a me ka glucose.Ua lilo ka pepa i substrate kūpono no ka microfluidics.Loaʻa i ka pepa nā waiwai e like me ka hydrophilicity a me ka porous structure, biocompatibility maikaʻi loa, ke kaumaha māmā, hiki ke hoʻololi, foldability, haʻahaʻa kumu kūʻai, maʻalahi o ka hoʻohana a me ka maʻalahi.Aia nā µPAD maʻamau i nā hale hydrophilic/hydrophobic i kūkulu ʻia ma luna o nā pani pepa.Ma muli o ke ʻano ʻekolu-dimensional, hiki ke hoʻokaʻawale ʻia nā μPAD i ʻelua-dimensional (2D) a me ʻekolu-dimensional (3D) μPADs.Hoʻokumu ʻia nā 2D µPAD ma ka hana ʻana i nā palena hydrophobic e hana i nā kahawai microfluidic, ʻoiai ʻo 3D µPAD e hana mau ʻia mai nā pūʻulu o nā papa o 2D microfluidic pepa, i kekahi manawa ma ka pelupe pepa, nā ʻenehana paheʻe, nā ala wehe, a me ka paʻi 3D [96].Hoʻomalu nui ʻia nā wai wai a i ʻole ke olaola ma ka μPAD e ka ikaika capillary me ka ʻole o kahi kumu mana o waho, e hoʻomaʻamaʻa i ka mālama mua ʻana o nā reagents, ka hoʻohana ʻana i nā laʻana, a me ka ʻike multiplex.Eia nō naʻe, ʻaʻole i lawa ka wikiwiki o ka ʻike ʻana, ka ʻike, a me ka hoʻohana hou ʻana [96, 127, 128, 129, 130].
Ma ke ʻano he kahua microfluidic maʻamau, ua hoʻolaha nui ʻia ʻo μPAD a hoʻomohala ʻia no ka maʻi molekala o nā maʻi maʻi e like me HCV, HIV, a me SARS-CoV-2 [131, 132].No ka ʻike koho a me ka maʻalahi o ka HCV, Tengam et al.[133] hoʻomohala i kahi biosensor hou e pili ana i ka pepa fluorescent me ka hoʻohana ʻana i kahi noiʻi nucleic acid kikoʻī e pili ana i ka pyrrolidinyl peptide.Nucleic acids ua covalently immobilized ma kahi hapa oxidized cellulose pepa e reductive alkylation ma waena o nā amino pūʻulu a me aldehyde pūʻulu, a me ka ike ma luna o fluorescence.Hiki ke heluhelu ʻia kēia mau hōʻailona e kahi hāmeʻa i hana kūikawā ʻia me kahi pahupaʻikiʻi fluorescent portable i hui pū ʻia me kahi pahupaʻikiʻi kelepona.Ma hope mai, ʻo Lu et al.[134] hoʻolālā i kahi electrode maʻalahi ma ka pepa e pili ana i ka nickel/gold nanoparticles/carbon nanotubes/polyvinyl alcohol organometallic framework composites no ka ʻike maka ʻana o ka maʻi HIV e ka DNA hybridization me ka methylene blue ma ke ʻano he hōʻailona DNA redox.I kēia mau lā, ʻo Chowdury et al.[135] hōʻike i kahi hoʻolālā platform hypothetical no ka hoʻāʻo ʻana i ka point-of-care µPAD me ka hoʻohana ʻana i ka wai maʻi maʻi i hui pū ʻia me LAMP a me ka ʻenehana kiʻi paʻi no ka ʻike analyte COVID-19.
Ke alakaʻi nei nā hoʻāʻo hoʻokahe ʻaoʻao i nā wai e nā ikaika capillary a mālama i ka neʻe ʻana o ka wai e ka pulu a me nā hiʻohiʻona o nā substrates porous a i ʻole microstructured.ʻO nā mea hoʻoheheʻe ʻaoʻao he laʻana, conjugate, incubator a me ka ʻike, a me nā pad absorbent.Hoʻomaopopo nā molekala nucleic acid i loko o ka LFA i nā mea hoʻopaʻa paʻa i mālama mua ʻia ma ke kahua paʻa a hoʻopaʻa ʻia e like me nā paʻakikī.I ka hele ʻana o ka wai i loko o ka incubation a me ka ʻike ʻana i nā papa, hopu ʻia nā mea paʻakikī e nā molekala hopu i loaʻa ma nā laina hoʻāʻo a me ka mana, e hōʻike ana i nā hopena hiki ke heluhelu pololei ʻia i ka maka ʻōlohelohe.ʻO ka maʻamau, hiki ke hoʻopau ʻia ka LFA i 2-15 mau minuke, ʻoi aku ka wikiwiki ma mua o ka ʻike kuʻuna.Ma muli o ke ʻano hana kūikawā, pono ka LFA i nā hana liʻiliʻi a ʻaʻole koi i nā mea hana hou aʻe, kahi e hoʻohana pono ai ia.He mea maʻalahi ka hana ʻana a me ka miniaturize, a ʻoi aku ka haʻahaʻa o ke kumukūʻai o nā substrate pepa.Eia nō naʻe, hoʻohana wale ʻia no ka loiloi qualitative, a paʻakikī loa ka ʻike ʻana i ka quantitative, a ua kaupalena ʻia ka mana multiplexing a me ka throughput, a hiki ke ʻike ʻia hoʻokahi wale nō nucleic acid i ka manawa [96,110,127].
ʻOiai ʻo ka hapa nui o nā noi o LFA e pili ana i nā immunoassays, ʻo ka hoʻohana ʻana i ka LFA no nā diagnostics molecular i nā microfluidic chips he maikaʻi a kaulana hoʻi [136].I ka hihia o ka maʻi hepatitis B, HIV a me SARS-CoV-2 LFA Gong et al.[137] hoʻolālā i kahi papa LFA nanoparticle hoʻololi a hōʻike i ka versatility o kēia miniaturized a me ka portable platform ma o ka ʻike makaʻala a me ka nui o nā pahuhopu lehulehu e like me ka HBV nucleic acid.Eia kekahi, ʻo Fu et al.Ua hōʻike ʻo [138] i kahi moʻolelo LFA e pili ana i ka spectroscopy Raman i hoʻonui ʻia no ka nānā ʻana o ka HIV-1 DNA ma nā haʻahaʻa haʻahaʻa.No ka ʻike wikiwiki a paʻakikī o SARS-CoV-2, Liu et al.[85] hoʻomohala i kahi ʻano hoʻoheheʻe kahe ʻana o RPA i hoʻohui ʻia i ka microfluidic ma o ka hoʻohui ʻana i ka RT-RPA a me kahi ʻōnaehana ʻike kahe holo ʻokoʻa āpau i loko o kahi ʻōnaehana microfluidic hoʻokahi.
ʻO ka hoʻohana ʻana i nā ʻano microfluidic platform ʻokoʻa ma muli o nā haʻawina kikoʻī, e lawe pono ana i nā hiki a me nā pono o nā paepae.Me nā valves kūpono, nā pāpaʻi a me nā ducts, ʻo LOCC ka paepae piha loa no ka noiʻi ʻokoʻa a me ka interoperability me ka lumi nui loa no ka hoʻomohala ʻana.No laila, manaʻolana mākou a manaʻo e hoʻokō ʻia nā haʻawina hou loa ma LOCC ma ke ʻano he hoʻāʻo mua a e hoʻokō ʻia nā kūlana.Eia kekahi, ʻoi aku ka maikaʻi a me ka pololei o nā ala e ʻike ʻia a hoʻohana ʻia i loko o ka ʻōnaehana.ʻOi aku ʻo LOAD i ka mana pololei o nā wai mai nā mea LOCC i loaʻa a hōʻike i nā pōmaikaʻi kūʻokoʻa i nā kaʻa hoʻokahi e ka ikaika centrifugal me ka ʻole o ka pono o nā kaʻa waho, ʻoiai hiki ke hoʻokaʻawale ʻia nā pane like.No laila, i ka wā e hiki mai ana, e lilo ʻo LOAD i kahua microfluidic nui me ka liʻiliʻi o nā hana lima a me nā ʻenehana ʻoi aʻe a me nā ʻenehana.Hoʻohui ka paepae µPAD i nā pono o LOCC a me nā mea i hoʻokumu ʻia i ka pepa no ke kumu kūʻai haʻahaʻa, hoʻohana hoʻokahi diagnostics.No laila, pono ka hoʻomohala ʻana i ka wā e hiki mai ana i nā ʻenehana kūpono a paʻa pono.Eia hou, ua kūpono ka LFA no ka ʻike maka ʻōlohelohe, e hoʻohiki ana e hōʻemi i ka hoʻohana ʻana i ka laʻana a me ka wikiwiki o ka ʻike.Hōʻike ʻia kahi hoʻohālikelike kahua kikoʻī ma ka Papa 2.
Hoʻokaʻawale nā kānana kikohoʻe i ka hāpana i loko o nā microreactors he nui, aia kēlā me kēia me kahi helu discrete o nā molekole i loaʻa [139, 140].Hāʻawi nā hōʻike kikohoʻe i nā pono koʻikoʻi no ka hana ʻana i ka helu ʻana ma o ka hana ʻana i nā kaukani o nā hoʻokolohua biochemical like i ka manawa like a me kēlā me kēia i loko o nā keʻena micron scale ma mua o ka manawa mau.Ke hoʻohālikelike ʻia me nā microfluidics kuʻuna, hiki i nā hopena o ke keʻena ke hoʻemi i ka nui o ka laʻana, hoʻonui i ka maikaʻi o ka hopena, a hiki ke hoʻohui maʻalahi me nā ʻano analytical ʻē aʻe me ka ʻole o ka pono o nā kahawai, nā pāpaʻi, nā pahu, a me nā hoʻolālā paʻa [141, 142, 143, 144, 145, 146, 147] .Hoʻohana ʻia nā ʻano hana ʻelua i nā hoʻokolohua kikohoʻe e loaʻa ai ka hoʻokaʻawale ʻokoʻa a me ka pololei o nā haʻina, me nā reagents a me nā laʻana e like me nā cell, nucleic acids, a me nā ʻāpana ʻē aʻe a i ʻole nā molekole: (1) hāʻule nā emulsions e hoʻohana ana i ka instability wai;(2) Hoʻokō ʻia ka mahele ʻana e nā palena geometric o ka hāmeʻa.Ma ke ʻano mua, hiki ke hana ʻia nā droplets i loaʻa nā reagents a me nā laʻana i nā microchannels e nā ʻano passive e like me ka co-current, crossflow, flow focusing, staged emulsification, microchannel emulsification, a me nā membrane ma o nā ikaika shear viscous a me ka emulsification me ka hoʻololi ʻana i ke kahawai.localization [143, 145, 146, 148, 149] a i ʻole ka hoʻohana ʻana i nā hana hana [150, 151], e hoʻolauna i ka ikehu hou ma o ka uila, magnetic, thermal and mechanical control.Ma ka hopena hope loa, ʻo ka like ʻana o ka nui o ka wai maikaʻi loa i loko o nā keʻena microfluidic e kaʻana like ʻia e ka mālama ʻana i nā hale spatial o ka nui like, e like me nā micropits a me nā papa kuhikuhi [152,153,154].ʻO ka mea nui, ʻo nā droplets nā ʻāpana kahe nui e hiki ke hana ʻia a hoʻohana ʻia ma luna o nā electrode arrays e pili ana i nā microfluidics digital (DMF).ʻO ka electrowetting o dielectrics kekahi o nā manaʻo DMF maikaʻi loa i aʻo ʻia, ʻoiai ʻo ka electrowetting o dielectrics e hiki ai i ka hoʻoponopono pololei ʻana i nā hāʻule o kēlā me kēia kanaka, e kāohi ana i ke ʻano o ka wai a me nā hōʻailona uila asymmetric e hele ana ma nā ʻaoʻao like ʻole [141, 144].ʻO nā hana koʻikoʻi me nā droplets i DMF e pili ana i ka hoʻokaʻawale ʻana, ka hoʻokaʻawale ʻana, a me ka hoʻohui ʻana [151, 155, 156], hiki ke hoʻohana ʻia i nā ʻano ʻano o ka nānā ʻana, ʻoi aku hoʻi i ka ʻike molecular [157, 158, 159].
ʻO ka ʻike ʻana i ka ʻakika nucleic kikohoʻe he ʻenehana diagnostic molecular kolu o ka hanauna ma hope o ka PCR maʻamau a me ka quantitative real-time PCR (qPCR), e like me ka hoʻopili kiʻekiʻe-throughput a me ka biopsy wai.I nā makahiki he ʻelua i hala iho nei, ua ulu wikiwiki nā ʻakika nucleic digital i ke kahua o ka diagnostics molecular of infectious pathogens [160, 161, 162].Hoʻomaka ka helu piha ʻana o ka ʻikena nucleic acid me ka hoʻopili ʻana i nā laʻana a me nā reagents i loko o nā keʻena hoʻokahi e hōʻoia i ka like o kēlā me kēia kaʻina pahuhopu ke komo i kēlā me kēia keʻena.Ma ka manaʻo, hiki ke hāʻawi ʻia i kēlā me kēia ʻāpana i nā kaʻina pahuhopu he nui, a i ʻole ʻaʻohe ʻōnaehana microreaction kūʻokoʻa.Ma o nā ʻano hana ʻike like ʻole i hōʻike ʻia ma luna nei, hiki ke ʻike ʻia nā keʻena me nā kaʻina microbial target e hoʻopuka i nā hōʻailona ma luna o kekahi paepae me ka maka ʻōlohelohe a i ʻole e ka mīkini a hōʻailona ʻia ma ke ʻano he maikaʻi, aʻo nā keʻena ʻē aʻe e hoʻopuka i nā hōʻailona ma lalo o ka paepae e hōʻailona ʻia he maikaʻi. .nā mea maikaʻi ʻole, e hoʻolilo i ka hōʻailona no kēlā me kēia ʻāpana he boolean.No laila, ma ka helu ʻana i ka helu o nā keʻena i hana ʻia a me ka nui o nā hopena maikaʻi ma hope o ka pane ʻana, hiki ke hoʻohālikelike ʻia nā kope kumu o nā hōʻike hoʻāʻo me ka hoʻohana ʻana i ke ʻano hoʻohele Poisson me ka ʻole o kahi pihi maʻamau, i koi ʻia no nā loiloi quantitative maʻamau e like me. e like me qPCR.[163] Hoʻohālikelike ʻia me nā ʻano diagnostic molecular kuʻuna, ʻoi aku ka kiʻekiʻe o ka ʻike ʻana i ka nucleic acid kikohoʻe, ʻoi aku ka wikiwiki o ka nānā ʻana a me ka naʻau, ka liʻiliʻi o nā reagents, ka liʻiliʻi o ka contamination, a me ka hoʻolālā a me ka hana maʻalahi.No kēia mau kumu, ua aʻo maikaʻi ʻia ka hoʻohana ʻana i nā hōʻike kikohoʻe, ʻoi aku ka nui o ka hoʻoheheʻe ʻana, no ka diagnostics molecular, ka hoʻohui ʻana i ka amplification a me nā ʻenehana heluhelu hōʻailona, i ka wā o ka maʻi koʻikoʻi o SARS-CoV-2.Eia kekahi laʻana,ʻo Yin et al.[164] hui pū ʻia nā ʻano kikohoʻe droplet a me ka PCR wikiwiki e ʻike i nā genes ORF1ab, N, a me RNase P i SARS-CoV-2 i loko o kahi puʻupuʻu microfluidic.ʻO ka mea nui, ua hiki i ka ʻōnaehana ke ʻike i kahi hōʻailona maikaʻi i loko o 115 kekona, ʻoi aku ka wikiwiki ma mua o ka PCR maʻamau, e hōʻike ana i kona pono i ka ʻike ʻana i ka point-of-care (Figure 7a).ʻO Dong et al.[165], Sow et al.[157], Chen et al.[166] a me Alteri et al.[167] hoʻohana pū kekahi droplet digital PCR (ddPCR) e ʻike iā SARS-CoV-2 i loko o kahi ʻōnaehana microfluidic me nā hopena hoihoi.No ka hoʻomaikaʻi hou ʻana i ka helu ʻike, ʻo Shen et al.[168] loaʻa i ka ddPCR-based chip imaging ma kahi liʻiliʻi e like me 15 s me ka hoʻohana ʻole ʻana i nā ʻenehana humuhumu kiʻi, e wikiwiki ana i ke kaʻina ʻenehana ddPCR mai ka lab a i ka noi.ʻAʻole hoʻohana wale ʻia nā ʻano hana hoʻonui wela e like me PCR, akā hoʻohana ʻia nā ʻano hana hoʻonui isothermal e hoʻomaʻamaʻa i nā kūlana hopena a me ka pane wikiwiki.Lu et al.[71] hoʻomohala i ka SlipChip no ka nānā ʻana i ka droplet, hiki ke hoʻohua i nā kulu o nā ʻano nui i nā kiʻekiʻe kiʻekiʻe i hoʻokahi ʻanuʻu a me ka helu ʻana i nā waikawa nucleic SARS-CoV-2 me ka hoʻohana ʻana i ka LAMP digital (Figure 7b).Ma ke ʻano he ʻenehana e ulu wikiwiki nei, hiki i ka CRISPR ke hana i kahi hana koʻikoʻi i ka ʻike ʻana i ka waika nucleic digital ma o ke kiʻi colorimetric maʻalahi me ka ʻole o ka pono o nā stains nucleic acid hou.ʻO Ackerman et al.ua hoʻomohala i ka hopena matrix combinatorial no ka loiloi multiplex o nā ʻakika nucleic.[158] ʻike ʻia he 169 mau maʻi e pili ana i ke kanaka, me ka SARS-CoV-2, i loko o nā droplets i loaʻa i ka CRISPR-Cas13-based nucleic acid detection reagents i kahi microwell assay (Figure 7c).Eia kekahi, hiki ke hoʻohana ʻia ka isothermal amplification a me ka ʻenehana CRISPR i ka ʻōnaehana hoʻokahi e hoʻohui i nā pono o nā mea ʻelua.Park et al.[169] Ua hoʻomohala ʻia kahi hōʻike kikohoʻe CRISPR/Cas12a i loko o kahi chip microfluidic pāʻoihana no ka ʻike ʻana i ka SARS-CoV-2 i hoʻopaʻa ʻia a me ka wela i hoʻokumu ʻia ma kahi pae RT-RPA hoʻokahi me kahi ʻike pōkole a ʻoi aku ka kiʻekiʻe. ratio manawa., ka laulā dynamic ākea a me ka naʻau maikaʻi (Fig. 7d).Hāʻawi ʻia kekahi mau wehewehe o kēia mau laʻana ma ka Papa 3.
ʻO ke kahua kikohoʻe maʻamau no ka ʻike waika nucleic.He ʻehā mau ʻanuʻu koʻikoʻi ka holo kaʻina hana PCR kikohoʻe wikiwiki: ka hoʻomākaukau ʻana i ka laʻana, ka hāʻawi ʻana i ka hui ʻana o ka hopena, ke kaʻina hana hoʻonui, a me ka helu ʻana (i hoʻololi ʻia mai [164]).b E hōʻike ana i ka nānā ʻana o nā kulu SlipChip no ka hoʻokumu ʻana o ka droplet ma ke kiʻekiʻe kiʻekiʻe (i hoʻololi ʻia mai [71]).c CARMEN-Cas workflow diagram13 (hoʻololi ʻia mai [158]).d Ka nānā ʻana o ka ʻike virus kikohoʻe holomua me CRISPR/Cas i loko o hoʻokahi ipu (i hoʻololi ʻia mai [169]).ʻO ka wai-i-ʻaila W/O, 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, severe acute respiratory syndrome, coronavirus 2 , RT Amplification o reverse transcriptase recombinase polymerase-RPA, S/B hōʻailona ma ke kua.
Ka manawa hoʻouna: Sep-15-2022
