Akụkọ

Usoro nyocha nke ọdịnala maka ịchọpụta ọrịa na-efe efe chọrọ iji ngwa benchtop na-adabaghị maka nnwale nleba anya (POCT).Microfluidics na-apụta bụ teknụzụ dị ntakịrị, akpaaka, yana agbakwunyere nke nwere ike ịbụ ụzọ ọdịnala maka nyocha ngwa ngwa, dị ọnụ ala, ziri ezi na saịtị.A na-eji usoro nyocha nke molecular eme ihe na ngwaọrụ microfluidic dịka ụzọ kachasị dị irè maka nchọpụta ọrịa.Nlebanya a na-achịkọta ọganihu na nso nso a na nyocha mkpụrụ ndụ ihe gbasara microfluidic nke ọrịa na-efe efe site na echiche agụmakwụkwọ na ụlọ ọrụ mmepụta ihe.Nke mbụ, anyị na-akọwa usoro nhazi nke nucleic acid na-adị na mgbawa, gụnyere pretreatment pretreatment, mmụba, na ịgụ mgbaàmà.A na-atụnyere njirimara, uru na adịghị ike nke ụdị anọ nke nyiwe microfluidic.Na-esote, anyị ga-atụle iji nyocha dijitalụ maka ọnụọgụ nucleic acid zuru oke.Achịkọta ngwaọrụ nyocha mkpụrụ ndụ ihe gbasara microfluidic nke oge gboo na nke na-adịbeghị anya dị ka ihe akaebe nke ọnọdụ ahịa ugbu a.N'ikpeazụ, anyị na-atụ aro ntụziaka n'ọdịnihu maka nchọpụta microfluidic nke ọrịa na-efe efe.
Ihe na-akpata ọrịa na-efe efe bụ ọrịa na-efe efe, gụnyere nje bacteria, nje, na nje nje ndị na-ekesa n'ụwa nile.N'adịghị ka ọrịa ndị ọzọ, nje nje na-ebute ngwa ngwa wee gbasaa n'etiti mmadụ na anụmanụ ndị ọbịa site na inoculation, mgbasa ozi ikuku na mmiri [1].Mgbochi ọrịa na-efe efe dị oke mkpa dị ka usoro ahụike ọha.Atụmatụ atọ bụ isi maka ịlụso ọrịa ọgụ ọgụ: (1) ịchịkwa isi iyi nke ọrịa;(2) nkwụsị nke ụzọ nnyefe;(3) nchebe nke ndị mmadụ nwere ike ime.N'ime atụmatụ ndị bụ isi, a na-ewere njikwa isi iyi nke ọrịa dịka atụmatụ kachasị mkpa n'ihi ịdị mma ya na ọnụ ala dị ala.Nchọpụta ngwa ngwa, ikewapụ, na ọgwụgwọ nke ndị butere ọrịa dị oke egwu, chọrọ usoro nyocha ngwa ngwa, nke nwere mmetụta na nke ziri ezi [2].Nchọpụta nke ugbu a nke ọrịa na-efe efe na-ejikọtakarị nyocha ụlọ ọgwụ dabere na akara na akara na nyocha ụlọ nyocha dị ka omenala cell na nchọpụta mkpụrụ ndụ, nke chọrọ ndị ọrụ a zụrụ azụ, usoro na-arụ ọrụ siri ike, na ngwa nyocha dị oke ọnụ [3, 4].Mgbochi nke ọrịa na-efe efe na-achọ nyocha ngwa ngwa, ọnụ ala na nke ziri ezi nke mpaghara, karịsịa na mpaghara ndị nwere oke akụrụngwa ebe ọrịa na-efe efe na-adịkarị ma dị njọ [5], yana ọgwụgwọ na ọzara ma ọ bụ n'ọgbọ agha, ebe ihe mberede na-enweghị atụ..nlekọta ahụike nwere oke [6].N'okwu a, microfluidics bụ teknụzụ na-ejikọta teknụzụ sistemụ microelectromechanical, nanotechnology, ma ọ bụ sayensị ihe maka ịmegharị mmiri nke ọma [7,8,9,10], na-enye ohere ọhụrụ maka nchọpụta nleba anya (POCT).) ndị na-efe efe n'èzí ụlọ ọgwụ na ụlọ nyocha.Tụnyere nyocha nke na-ewe oge ọdịnala, teknụzụ microfluidic na-enye nlele na nchekwa ego maka nyocha mkpụrụ ndụ n'oge ntiwapụ ọrịa.Mgbasa zuru ụwa ọnụ nke ọrịa coronavirus 2019 (COVID-19) bụ ihe na-akpata nnukwu ọrịa iku ume iku ume coronavirus 2 (SARS-CoV-2), yabụ na-ekwusi ike ọzọ mkpa microfluidics maka mgbochi oge na njikwa nke ọrịa a [11, 12] , 13].N'adịghị ka nchọpụta ọdịnala, microfluidic POCT na-eji obere ngwaọrụ ndị a na-ebugharị ebugharị sitere na ndị nyocha benchtop ruo obere mpempe ule sidestream iji nwalee nso ebe nlele [14].Nnwale ndị a na-egosipụta nkwadebe dị mfe ma ọ bụ enweghị nlere anya, nkwalite mgbaama ngwa ngwa, yana ọgụgụ mgbaama na-enwe mmetụta nke na-ebute obere oge na nsonaazụ ziri ezi n'ime nkeji.Nnweta na mmepụta oke nke ngwa nlekọta ahụike nke microfluidic agbasawanyela ngwa nyocha ha dị ọnụ ahịa na kpọmkwem n'èzí ụlọ ọgwụ, nso onye ọrịa, na ọbụna n'ụlọ.
N'ime atụmatụ ndị dị ugbu a maka ịchọpụta ọrịa na-efe efe, nchọpụta mkpụrụ ndụ ihe nketa bụ otu n'ime ihe kacha nwee mmetụta [15, 16].Na mgbakwunye, a na-ejikarị nyocha nke mkpụrụ ndụ ihe dị ka ọkọlọtọ ọla edo maka nchọpụta COVID-19 na-aga n'ihu, na-enye ohere ịchọpụta kpọmkwem mpaghara nje nke RNA ma ọ bụ DNA tupu mmalite nke nzaghachi mgbochi [17, 18].Na nyochaa ugbu a, anyị na-eweta ọganihu kachasị ọhụrụ na usoro nchọpụta ihe omimi nke microfluidics maka ọrịa ndị na-efe efe, site n'echiche agụmakwụkwọ na echiche ụlọ ọrụ n'ọdịnihu (Fig 1).Anyị ga-amalite site na isi nzọụkwụ atọ na nchọpụta nucleic acid: pretreatment on-chip sample pretreatment, nucleic acid amplification, na ịgụ akara.Anyị wee tụnyere ụdị dị iche iche nke microfluidic nyiwe na nhazi na ọrụ ha, na-egosi njirimara pụrụ iche (ike na adịghị ike).A tụlere n'ihu na nchọpụta nucleic acid dijitalụ ma nye ya dị ka ọmụmaatụ nke teknụzụ ọgbọ nke atọ maka ọnụọgụ zuru oke nke ụmụ irighiri ihe na-efe efe.Na mgbakwunye, a ga-egosipụta ọtụtụ ngwa ahịa POCT na-ahụkarị na nke kachasị ọhụrụ iji gosipụta ọnọdụ ahịa microfluidic POCT dị ugbu a maka nyocha mkpụrụ ndụ.Anyị ga-atụle ma kọwaa ọhụụ anyị maka ngwa n'ọdịnihu.
Enwere ike kewaa modul nke ibe microfluidic maka nchọpụta nucleic acid ụzọ atọ (nlekọta, nnabata, na akara) dịka ọrụ ha si dị [19].N'ime modul ndị a, modul nlere anya na-achọpụta lysis sample na mmịpụta nucleic acid.Modul ihe mmetụta na-achịkwa ntụgharị na mmụba nke akara nucleic acid.Igwe nrịbama na-achọpụta mgbama tụgharịrị ma hazie ya site na modul mmetụta.Dabere na usoro nke ịchọpụta nucleic acids na mgbawa, anyị ga-achịkọta ibe dị iche iche nke nwere ike ịghọta ọrụ "ntinye na mmepụta".
Nzọụkwụ mbụ na nchọpụta nucleic acid bụ mmịpụta nucleic acid, ya bụ ikewapụ acid nucleic e lekwasịrị anya na nlele mbụ.A na-eme mmịpụta nucleic acid iji sachaa acids nucleic site na mmetọ ụmụ irighiri ihe ndị ọzọ, hụ na iguzosi ike n'ezi ihe nke usoro izizi nke mkpụrụ ndụ nucleic acid, wee bulie nsonaazụ ya.Mwepụta nucleic acid chọrọ nyocha lysis dị mkpa na njide nucleic acid, ịdị mma na arụmọrụ ya nwere nnukwu mmetụta na nyocha na nsonaazụ nyocha.Mmetụta ọ bụla dị nro n'oge mmịpụta nwere ike igbochi nchọpụta ọzọ.Dịka ọmụmaatụ, mmeghachi omume nke polymerase chain (PCR) na loop isothermal amplification (LAMP) na-egbochi ụfọdụ ihe mgbaze organic fọdụrụnụ dị ka ethanol na isopropanol na nucleic acid reagents reagents [20].Mwepụta mmiri mmiri na mmịpụta nke siri ike bụ ụzọ kachasị ewu ewu maka ikewapụ acid nucleic [21], agbanyeghị, mmịpụta mmiri mmiri na mgbawa nwere oke oke, ebe ọ bụ na reagents eji na mmịpụta mmiri mmiri na-akpata corrosion nke ọtụtụ ibe microfluidic. .N'ebe a, anyị na-akọwapụta usoro mmịpụta usoro siri ike dabere na microarray wee tulee uru na ọghọm ha.
Silicon bụ ihe eji eme ihe kwekọrọ na acid nucleic n'ihi nkwụsi ike ya, nkwụsi ike ya na ịdị mfe nke mgbanwe [22].N'ụzọ dị mkpa, mgbe ejiri silica ma ọ bụ ihe ndị ọzọ gbanwee, ihe mejupụtara a na-egosipụta ihe onwunwe iji gbasaa acid nucleic na-adịghị mma n'okpuru pH dị ala, ọnọdụ nnu dị elu ka ọ na-eji pH dị elu, ngwọta nnu dị ala.Dabere na ihe omume a, ọ ga-ekwe omume ime ka acid nucleic dị ọcha.
A na-eji ụdị dị iche iche nke silica eme ihe maka mmịpụta nucleic acid na microfluidics, dị ka beads silica, powders, microfiber filter, na silica membranes [23, 24, 25, 26].Dabere na njirimara nke ihe ahụ, a pụrụ iji ihe ndị dabeere na silicon mee ihe na microcircuits n'ụzọ dị iche iche.Dịka ọmụmaatụ, silica granules, powders, na nanofilters azụmahịa nwere ike itinye n'ime pores ma ọ bụ microchannels nke microfluidic ibe ma nyere aka wepụ nucleic acids na sample [27, 28, 29].A nwekwara ike iji akpụkpọ anụ silica megharịrị ihu iji mee ka DNA dị ọcha ngwa ngwa site na nje na-efu ọnụ.Dịka ọmụmaatụ, Wang et al.[30] Site n'ijikọta mmeghachi omume nkwuwanye ụda na mgbanwe yinye nke vesicle na-agbaziri agbaziri na membranes silica nke ejiri chitosan oligosaccharides kpuchie, ewebata sistemu enwere ike ibugharị nke achọpụtara 102-108 colony na-emepụta nkeji nke ọma.(CFU)/ml Vibrio parahaemolyticus., na ọnụnọ nke nje na-adị mfe ịhụ.Powell et al.[31] Mgbe ahụ, a na-eji microarrays dabeere na silicon chọpụta nje ịba ọcha n'anya C (HCV), nje immunodeficiency virus (HIV), nje Zika, na papillomavirus mmadụ na mgbasa ozi na-akpaghị aka, bụ nke e mepụtara 1.3 μl microreactor tortuous iji jide nje RNA.ma rụọ ọrụ nkwalite.Na mgbakwunye na usoro ndị a, microcolumns silica ndị na-emezigharị n'elu na-arụkwa ọrụ dị mkpa na mmịpụta nucleic acid, n'ihi na geometry na njirimara nke ihe na-agbanwe agbanwe na-abawanye arụmọrụ mmịpụta nke ukwuu.Chen et al.[32] tụpụtara ikpo okwu microfluidic maka ikewapụ RNA nwere obere uche dabere na microcolumn silicon nwere mkpuchi amino.Ngwa microfluidic a na-ejikọta ọtụtụ micropillars 0.25 cm2 na mkpụrụ silicon iji nweta arụmọrụ mmịpụta dị elu site na mpaghara elu dị elu na nha nha nha.Uru nke imewe a bụ na ngwaọrụ microfluidic nwere ike nweta ihe ruru 95% nucleic acid mmịpụta arụmọrụ.Usoro ndị a dabere na silicon na-egosipụta uru nke ikewapụ acid nucleic ngwa ngwa na ọnụ ala dị ala.Na mgbakwunye na ibe microfluidic, usoro mmịpụta nke silicon nwere ike ọ bụghị naanị ịbawanye arụmọrụ nke nchọpụta nucleic acid, kamakwa ọ na-akwado miniaturization na ntinye nke ngwaọrụ nyocha [20].
Ụzọ nkewa magnetik na-eji irighiri ihe ndọta kewapụ acid nucleic n'ihu oghere ndọta dị n'èzí.Ihe ndọta a na-ejikarị gụnyere Fe3O4 ma ọ bụ γ-Fe2O3 ihe ndọta ejiri silica, amino na carboxyl kpuchie [33,34,35,36].Ihe dị iche iche nke ihe ndọta ma e jiri ya tụnyere usoro SPE dabeere na silicon bụ mfe nke nhazi na njikwa na magnets mpụga.
Iji electrostatic mmekọrịta n'etiti nucleic acid na silica, n'okpuru ọnọdụ nke elu nnu na ala pH, nucleic acids na-adsorbed n'elu silica-kpuchie magnetik ahụ, ebe n'okpuru ọnọdụ nke obere nnu na elu pH, ụmụ irighiri ihe nwere ike ịsacha ụmụ irighiri ihe. ọzọ..Mpempe ndọta nke silica nwere mkpuchi na-eme ka o kwe omume iwepụta DNA site na nnukwu ihe nlele (400 μL) site na iji mmegharị magnetik na-achịkwa [37].Dị ka ihe ngosi, Rodriguez-Mateos et al.[38] na-eji magnets tunable iji chịkwaa nnyefe nke bead magnet n'ime ụlọ dị iche iche.Dabere na ụmụ irighiri ihe ndọta nke silica nwere mkpuchi, 470 mbipụta / mL nke SARS-CoV-2 genomic RNA nwere ike ịwepụta ya na nlele mmiri mkpofu maka nchọpụta ntụgharị ntụgharị LAMP (RT-LAMP) na enwere ike ịgụ azịza ya n'ime elekere 1.anya gba ọtọ (Fig 2a).
Ngwaọrụ dabere na ihe ndọta na porous.Eserese echiche nke ngwaọrụ microfluidic IFAST RT-LAMP maka nchọpụta SARS-CoV-2 RNA (emepụtara na [38]).b Centrifugal Micro ngwaọrụ maka dSPE nke buccal swab nucleic acid (nke sitere na [39]).c Ihe nleba anya nleba anya nke ejiri aka ya rụọ n'ime ya na-eji kaadị FTA® (emetere na [50]).d Fusion 5 akwụkwọ nzacha ejiri chitosan gbanwee (emetere na [51]).SARS-CoV-2 siri ike ọrịa iku ume iku ume coronavirus 2, RT-LAMP reverse transcription loop mediated isothermal amplification, FTA na-achọpụta teknụzụ, NA nucleic acid.
Ụmụ irighiri ihe ndọta chaja nke ọma dị mma maka ijikọta ọkpụkpụ azụ phosphate nke nucleic acid.N'otu oge ntinye nnu, otu phosphate na-adịghị mma nke nucleic acid nwere ike ịgbanye nke ọma n'elu ihe mejupụtara ihe ndọta.Ya mere, e mepụtara nanoparticles magnetik nwere elu siri ike yana nnukwu njupụta nke otu amino maka mmịpụta nke acid nucleic.Mgbe nkewa na igbochi magnetik, enwere ike iji magnetik nanoparticles na ogige DNA mee ihe na PCR, nke na-ewepụ mkpa maka ịdị ọcha dị mgbagwoju anya na nke na-ewe oge na ọrụ elution [35].A na-ejikwa nanoparticles magnetik kpuchie na otu carboxyl na-adịghị mma iji kewapụ acids nucleic na-etinye n'elu elu nke polyethylene glycol na ngwọta sodium chloride [36].Site n'elu bead magnetik emezigharịrị n'elu, mmịpụta DNA dakọtara na mmụba na-esote.Dignan et al.[39] kọwara ikpo okwu centrifugal microfluidic akpaghị aka na obere ibugharị maka pretreatment acid nucleic, na-enye ndị ọrụ na-abụghị teknụzụ ohere iji ya na saịtị.Tụkwasị na nke ahụ, ndakọrịta nke DNA dịpụrụ adịpụ na LAMP, usoro nke dabara nke ọma maka nyocha nucleic acid na-elekọta, na-egosipụtakwa ihe ndị dị ntakịrị na-achọ ngwá ọrụ na ihe kwesịrị ekwesị maka nyocha colorimetric (Fig 2b).
Ụzọ bead magnetik na-enye ohere nke mmịpụta na-akpaghị aka, ụfọdụ n'ime ha dị na ndị na-ewepụta acid nucleic na-emepụta ihe [KingFisher;ThermoFisher (Waltham, MA, USA), QIAcube® HT;CapitalBio (Beijing, China) na Biomek®;Beckman (Miami, USA).), Florida, USA).Enwere ike iji uru nke ijikọta beads magnetik na microfluidics maka mmịpụta nucleic acid nke ọma, nke nwere ike ịkwalite mmepe nke nchọpụta mkpụrụ ndụ;Otú ọ dị, nchikota nke magnetik beads na microfluidics ka na-adabere kpamkpam na mgbagwoju njikwa usoro maka kpọmkwem mkpachapụ nke magnetik beads, nke na-akọwa na ewu ewu nke azụmahịa ngwaahịa ịbụ nnukwu na oké ọnụ, nke na-amachi n'ihu ngwa nke magnetik beads na POCT.
A na-ejikwa ọtụtụ ihe ndị nwere porous dị ka ihe nzacha nitrocellulose gbanwetụrụ, kaadị Finders Technology Associates (FTA), akwụkwọ nzacha nke polyethersulfone, na ihe mkpuchi glycan maka nchọpụta nucleic acid [40, 41, 42, 43, 44].A na-ebu ụzọ jiri ihe ndị nwere fibrous dị ka akwụkwọ fibrous were kewapụ DNA site na iji eriri jikọtara ụmụ irighiri ihe DNA nwere ogologo ogologo.Obere oghere na-eduga na mmachi anụ ahụ siri ike nke mkpụrụ ndụ DNA, nke na-emetụta mmịpụta DNA nke ọma.N'ihi nha pore dị iche iche nke akwụkwọ fibrous, arụmọrụ mmịpụta enweghị ike izute mkpa nke mmụba DNA [45, 46].Kaadị FTA bụ akwụkwọ nzacha azụmaahịa nke a na-eji n'ọhịa ọgwụ nyocha ma na-ejikwa ya n'akụkụ ndị ọzọ nke nyocha mkpụrụ ndụ.Site na iji akwụkwọ nzacha cellulose nke nwere kemịkalụ dị iche iche na-eme ka akpụkpọ anụ ahụ dị n'ime ihe nlele ahụ, a na-echebe DNA a tọhapụrụ pụọ na mmebi ruo afọ 2.N'oge na-adịbeghị anya, e mepụtala akwụkwọ cellulose nke etinyere n'ime maka nchọpụta mkpụrụ ndụ nke nje dị iche iche, gụnyere SARS-CoV-2, leishmaniasis, na ịba [47,48,49].HIV na plasma dịpụrụ adịpụ na-edobere ya ozugbo, na viral nucleic acid na-eme ka ọ dịkwuo mma na akpụkpọ anụ FTA® nke e wuru n'ime concentrator, nke na-enye ohere ịmepụta nucleic acid nke ọma [50] (Fig 2c).Nsogbu bụ isi na nchọpụta nucleic acid site na iji kaadị FTA bụ na kemịkalụ dị ka guanidine na isopropanol na-egbochi mmeghachi omume mmụba na-esote.Iji dozie nsogbu a, anyị mepụtara Fusion 5 chitosan-modified paper filter, nke na-ejikọta uru nke ma njikọ anụ ahụ nke mkpụrụ ndụ DNA na akwụkwọ nzacha fibrous, yana mgbasa ozi electrostatic nke DNA na ogige chitosan gbanwetụrụ iji nweta mmịpụta nucleic acid dị mma nke ukwuu. ..nzacha eriri [51] (Fig. 2d).N'otu aka ahụ, Zhu et al.[52] gosipụtara usoro PCR nke chitosan gbanwetụrụ dabere na sistemu microfluidic capillary in situ maka ikewapụ ngwa ngwa na ịchọpụta nje Zika RNA.Enwere ike ịtinye acid nucleic n'ime ngwakọta lysate/PCR agwakọtara, dabere na mgbanyụ ma gbanyụọ nke chitosan.gbanye na gbanyụọ", na-anabata pH.
Dị ka ekwuru n'elu, atụmatụ ndị a na-ejikọta uru nke ihe dị iche iche siri ike na-emepụta ma na-abawanye arụmọrụ nke mmịpụta nucleic acid na microfluidics.N'ime ngwa bara uru, iji ihe ndị a eme ihe n'ọtụtụ buru ibu bụ enweghị ego, na ọgwụgwọ elu ma ọ bụ ngbanwe elu nke ihe ndị a na-ahụkarị na ihe ndị a nwekwara ike ichekwa ọrụ ha.Ya mere, a kwenyere na mmejuputa atumatu ndị a mgbe nyochachara ụgbọelu nwere ike ibelata ọnụ ahịa.
Nnwale nucleic acid na nyiwe microfluidic na-ejikarị obere ihe nlele (<100 µl), yabụ na-achọ mmụba nke acid nucleic lekwasịrị anya na nyocha pụrụ iche maka ntụgharị gaa na mgbama dabara adaba maka nchọpụta ala (optical, eletriki na magnetik) [53, 54]. Nnwale nucleic acid na nyiwe microfluidic na-ejikarị obere ihe nlele (<100 µl), yabụ na-achọ mmụba nke acid nucleic lekwasịrị anya na nyocha pụrụ iche maka ntụgharị gaa na mgbama dabara adaba maka nchọpụta ala (optical, eletriki na magnetik) [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]. Mgbe a na-anwale acids nucleic na nyiwe microfluidic, a na-ejikarị obere ihe nlele (<100 µL) eme ihe, yabụ achọrọ mmụba nke acid nucleic lekwasịrị anya nwere nyocha pụrụ iche iji gbanwee ya ka ọ bụrụ akara dabara adaba maka nchọpụta na-esote (optical, eletriki na magnetik) [53, 54].平台 上 上 的 的 使用 使用 使用 使用 使用 μ μ μ 需要 需要 转换 转换 转换 转换 转换 下游 下游 下游 下游 和 和 和 和 [53, 54 ]平台 上 上 上 的 检测 使用 使用 使用 使用 μl), 因此 μ μ 扩增 转换 转换 转换 下游 下游 (光学, 磁学 [53, 54, 54) ] 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]]. Nchọpụta nke nucleic acids na microfluidic nyiwe na-ejikarị obere ihe nlele (<100 μl), nke na-achọ mmụba nke acid nucleic lekwasịrị anya na nyocha pụrụ iche iji mee ka ha ghọọ akara maka nchọpụta na-esote (optical, electric, and magnet) [53, 54] .Nkwalite nucleic acid na microfluidics nwekwara ike ime ka mmeghachi omume dị ngwa, bulie oke nchọta, belata ihe nlele chọrọ, ma kwalite izi ezi nchọpụta [55, 56].N'ime afọ ndị na-adịbeghị anya, site n'ịghọta nchọpụta ngwa ngwa na nke ziri ezi, a na-etinye ụzọ dị iche iche nucleic acid amplification na microfluidics, gụnyere PCR na ụfọdụ mmeghachi omume amplification isothermal.Akụkụ a ga-achịkọta ụzọ maka nchọpụta nucleic acid dabere na sistemụ microfluidic.
PCR bụ ịme anwansị nke usoro mmegharị DNA nke akụkụ ahụ, nke akọwara n'ụzọ zuru ezu n'ebe ndị ọzọ na agaghị atụle ya ebe a.PCR nwere ike ịbawanye ntakịrị DNA/RNA e lekwasịrị anya n'ọ̀tụ̀tụ̀ dị mkpirikpi, na-eme PCR ka ọ bụrụ ngwá ọrụ dị ike maka nchọpụta ngwa ngwa nke nucleic acid.N'ime iri afọ ndị na-adịbeghị anya, e mepụtala ọtụtụ ngwaọrụ microfluidic obere obere nwere sistemu ịgba ígwè PCR iji gboo mkpa nke nchọpụta nleba anya [57, 58].Enwere ike kewaa PCR on-chip n'ụdị anọ (usoro omenala, na-aga n'ihu na-aga n'ihu, na-atụgharị oghere, na PCR convective) dịka usoro nchịkwa okpomọkụ dị iche iche [59].Dịka ọmụmaatụ, Gee et al.[60] mepụtara usoro PCR (RT-qPCR) kpọmkwem n'usoro microfluidic nke onwe ha maka nchọpụta multiplex SARS-CoV-2, nje influenza A na B na akpịrị swab sample (Fig 3a).Park et al.[61] wuru mgbawa nyocha pathogen dị mfe site na ijikọ ihe nkiri PCR dị mkpa, electrodes, na modul microfluidic polydimethylsiloxane na-arụ ọrụ.Agbanyeghị, ọrụ abụọ a gụnyere mmejọ nkịtị nke PCR.PCR chọrọ ịnya igwe ọkụ, nke na-egbochi mbelata ngwaọrụ yana belata oge nnwale.
Mmepe nke microfluidic na-aga n'ihu na-aga n'ihu na PCR na-agbanwe oghere dị oke mkpa iji dozie okwu a.Iji ogologo ọwa serpentine ma ọ bụ ọwa dị mkpụmkpụ kwụ ọtọ, PCR na-aga n'ihu nwere ike inye mmụba ngwa ngwa site na ịgbasa reagents na mpaghara preheat atọ nwere mgbapụta mgbapu.Ọrụ a na-ezere oge mgbanwe n'etiti okpomọkụ mmeghachi omume dị iche iche ma si otú a na-ebelata oge ule nke ọma [62] (Fig 3b).Na nyocha ọzọ Jung et al.[63] tụpụtara ọhụrụ rotary PCR mkpụrụ ndụ ihe nketa analyzer nke na-agwakọta àgwà nke ofu na eruba PCR maka ultrafast na multiplex reverse transcription PCR (Fig. 3c).Maka mmụba nke acid nucleic, PCR microchip ga-atụgharị site na mgbochi ọkụ atọ na okpomọkụ dị iche iche: 1. Denaturation block 94°C, 2. Annealing block na 58°C, 3. Mgbasawanye mgbochi na 72°C.
Ngwa nke PCR na microfluidics.Ihe nnochite anya nke dirRT-qPCR n'elu ikpo okwu microfluidic (nke sitere na [60]).b Ihe nlere anya nke microarray PCR na-aga n'ihu dabere na ọwa serpentine (nke sitere na [62]).c Nnọchite anya ihe gbasara mkpụrụ ndụ ihe nketa PCR rotary, nke nwere microchip, mgbochi ọkụ atọ na moto stepper (emetere na [63]).d Eserese nke PCR thermoconvection nwere centrifugation na ntọlite ​​​​(emepụtara na [64]).DirRT-qPCR, mmeghachi omume ntụgharị ntụgharị polymerase ozugbo
N'iji capillaries na loops ma ọ bụ ọbụna efere dị gịrịgịrị, convection PCR nwere ike ịbawanye ngwa ngwa nucleic acid site na convection na-ekpo ọkụ n'efu na-enweghị mkpa maka mgbapụta mpụga.Dịka ọmụmaatụ, e mepụtara ikpo okwu olefin polymer microfluidic cyclic na usoro ikpo ọkụ na-agbanwe agbanwe nke na-eji ịgba ígwè ọkụ na centrifugation na PCR loop microchannel [64] (Fig 3d).Ngwọta mmeghachi omume na-ebute site na convection thermal, nke na-aga n'ihu na-agbanwe oke ma dị ala na microchannel nwere usoro afọ.Enwere ike mezue usoro nkwalite ahụ na nkeji iri na njedebe nke 70.5 pg / ọwa.
Dị ka a tụrụ anya ya, PCR ngwa ngwa bụ ngwá ọrụ dị ike maka nyocha ihe nleba anya n'ụzọ zuru ezu na usoro nyocha multiplex.PCR ngwa ngwa na-ebelata oge achọrọ iji chọpụta SARS-CoV-2, nke na-enye aka na njikwa dị mma nke ọrịa COVID-19.
PCR chọrọ igwe igwe na-ekpo ọkụ dị mgbagwoju anya nke na-adabaghị maka POCT.N'oge na-adịbeghị anya, etinyere usoro nkwalite isothermal na microfluidics, gụnyere ma ọnweghị oke na LAMP, recombinase polymerase amplification (RPA), na nkwalite dabere na usoro nucleic acid [65,66,67,68].Site na usoro ndị a, a na-amụba acids nucleic na okpomọkụ na-adịgide adịgide, na-eme ka ịmepụta ngwaọrụ POCT dị ọnụ ala, na-enwe mmetụta dị ukwuu maka nchọpụta molecular.
Nnwale LAMP nke dabeere na microfluidics dị elu na-enye ohere ịchọta ọtụtụ ọrịa na-efe efe [42, 69, 70, 71].Na mgbakwunye na sistemu microfluidic centrifugal, LAMP nwere ike imeziwanye ngwa ngwa nke nchọpụta nucleic acid [69, 72, 73, 74, 75].Emepụtara SlipChip spin-na-react maka nhụta anya nke ọtụtụ nje bacteria na-eji LAMP [76] (Fig. 4a).Mgbe ị na-eji LAMP kachasị mma na nyocha ahụ, ogo mgbama fluorescence na mkpọtụ dị ihe dịka okpukpu ise, na njedebe nchọta ruru 7.2 mbipụta/μl nke DNA genomic. Ọzọkwa, a na-ahụ anya ịdị adị nke nje nje nje nje iri nri ise, gụnyere Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis na Vibrio parahaemolyticus, dabere na usoro na <60 min. Ọzọkwa, a na-ahụ anya ịdị adị nke nje nje nje nje iri nri ise, gụnyere Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis na Vibrio parahaemolyticus, dabere na usoro na <60 min.Ọzọkwa, a na-ahụ anya ọnụnọ nke nje nje nje ise nke tract digestive, gụnyere Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvialis na Vibrio parahaemolyticus site na iji usoro a n'ihe na-erughị nkeji 60., 基于 该 该 <60 分钟 分钟 分钟 分钟 分钟 分钟 分钟 分钟 分钟 分钟 分钟 内 内 了 了 了 了 了 分钟 了 五 分钟 了 可 了 分钟 了 分钟 五 种 五 五 五 消化道 细菌病 细菌病 消化道 消化道 蜡状 存在 存在, 肠杆菌 肠杆菌, 肠杆菌 沙门 沙门 沙门 弧菌 弧菌 和 和 弧菌., 基于 该 该 该 <60 分钟 了 分钟 分钟 分钟 分钟 分钟 了 分钟 五 了 分钟 分钟 分钟 了 视化 五 五 五 种 消化道 芽孢杆菌, 芽孢杆菌, 芽孢杆菌, 肠杆菌 肠杆菌, 氏 和 副溶血 性 性 弧菌弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 弧菌 HIPNa mgbakwunye, a na-ahụ anya ọnụnọ nke nje nje nje eriri afọ ise, gụnyere Bacillus cereus, Escherichia coli, Salmonella enterica, Vibrio fluvius, na Vibrio parahaemolyticus, site na iji usoro a n'ihe na-erughị nkeji 60.
Uru nke LAMP na microfluidics gụnyere, n'etiti ndị ọzọ, nzaghachi ngwa ngwa na nchọpụta ntakịrị.Otú ọ dị, n'ihi okpomọkụ mmeghachi omume (ihe dị ka 70 Celsius C), aerosols bụ ihe a na-apụghị izere ezere na-emepụta n'oge LAMP, na-ebute ọnụego ụgha dị elu.Assay nkọwapụta, nhazi primer, na njikwa okpomọkụ dịkwa mkpa ka emeziwanye ya maka LAMP.Na mgbakwunye, atụmatụ mgbawa na-emejuputa ọtụtụ nchọpụta ebumnuche na otu mgbawa bara nnukwu uru na ekwesịrị imepụta ya.Na mgbakwunye, LAMP dabara maka nchọpụta ọtụtụ ebumnuche agbakwunyere na otu mgbawa, nke dị oke mkpa, mana a ka nwere ọtụtụ ohere maka mmepe.
Enwere ike iji RPA belata ọnụego ụgha ụgha dị elu nke LAMP, n'ihi na ọnọdụ mmeghachi omume dị ntakịrị (~ 37 Celsius) na-ebute nsogbu evaporation dị ole na ole [77].Na usoro RPA, abụọ na-emegide primers na-amalite njikọ DNA site na ijikọ na recombinase na mmụba nwere ike mechaa n'ime nkeji iri [78,79,80,81].Ya mere, usoro RPA dum dị ngwa ngwa karịa PCR ma ọ bụ LAMP.N'ime afọ ndị na-adịbeghị anya, egosiri na nkà na ụzụ microfluidic na-emewanyewanye ọsọ na izi ezi nke RPA [82,83,84].Dịka ọmụmaatụ, Liu et al.[85] mepụtara microfluidic integrated lateral flow polymerase recombinase amplification assay maka nchọpụta ngwa ngwa na nke siri ike nke SARS-CoV-2 site na ijikọ ntụgharị ntụgharị RPA (RT-RPA) na sistemu nyocha n'akụkụ ihu igwe.n'ime otu microfluidic usoro.Ọgụgụ 4b).Oke nchọpụta bụ 1 oyiri/µl ma ọ bụ 30 nnomi/nlekọta, enwere ike mechaa nchọpụta n'ime ihe dịka nkeji iri atọ.Kong et al.emebela ngwaọrụ microfluidic wearable.[86] ji okpomọkụ ahụ na usoro nchọpụta fluorescence dabeere na ekwentị mkpanaaka iji chọpụta DNA HIV-1 ngwa ngwa na iji RPA (Njirimara 4c).Nyocha RPA wearable na-achọpụta 100 nnomi/mL nke usoro ebumnuche n'ime nkeji iri abụọ na anọ, na-egosipụta nnukwu ikike maka nyocha ngwa ngwa nke ụmụ ọhụrụ bu nje HIV-1 na ntọala nwere oke akụrụngwa.
Isothermal amplification na nleba anya nlekọta (POCT).Mmepe na mmepụta nke spin na mmeghachi omume SlipChip.Mgbe ịgbado ọkụ plasma gasịrị, a na-achịkọta ibe n'elu na nke ala ya na mkpụrụ osisi ka ọ bụrụ mgbawa ikpeazụ (nke sitere na [76]).b Schematic nke usoro MI-IF-RPA maka nchọpụta COVID-19 (emepụtara na [85]).c Schematic nke ule RPA wearable maka nchọpụta ngwa ngwa nke DNA HIV-1 (emetere na [86]).SE Salmonella enterica, VF Vibrio fluvius, VF Vibrio fluvius, VP Vibrio parahaemolyticus, BC Bacillus cereus, EC Escherichia coli, FAM carboxyfluorescein, nje immunodeficiency mmadụ HIV, RPA recombinase polymerase amplification, LED ìhè emitting diode, MI-IF-RPA Microfluidics Integrated Polyfluids. Mmụba
RPA dabere na Microfluidic na-etolite ngwa ngwa, agbanyeghị, ọnụ ahịa nrụpụta mgbawa na oriri mmeghachi omume dị oke oke na a ga-ebelatarịrị iji bulie nnweta teknụzụ a.Tụkwasị na nke ahụ, mmetụta dị elu nke RPA nwere ike imetụta mmụba nke ngwaahịa ndị na-abụghị nke a kapịrị ọnụ, karịsịa na ọnụnọ nke mmetọ.Mmachi ndị a nwere ike imetụta ngwa RPA na sistemu microfluidic yana uru n'ihu n'ihu.Achọkwara primers na nyocha emebere nke ọma maka ebumnuche dị iche iche iji melite ike nke atụmatụ microfluidic dabere na RPA na POCT.
Cas13 na Cas12a nwere ikike nkewapụ acids nucleic na-enweghị usoro wee si otú a mepụta dị ka nchọpụta na ngwaọrụ nyocha.Cas13 na Cas12a na-arụ ọrụ mgbe ejikọtara ya na DNA ma ọ bụ RNA, n'otu n'otu.Ozugbo etinyere ya, protein ahụ na-amalite ịgbajisị acids nucleic ndị ọzọ dị nso, mgbe nke ahụ gasịrị, ntuziaka RNA ndị na-elekwasị anya n'ụdị nucleic acid pụrụ iche nwere ike kụwaa nyocha fluorescent menyụọ ma hapụ fluorescence.Dabere na tiori a, Kellner et al.[87] mepụtara usoro dabere na Cas13 [Specific High-sensitivity Enzymatic Reporter UnlockING (SHERLOCK)], na Broughton et al.[88] mepụtara ụzọ ọzọ dabere na Cas12a [CRISPR Trans Reporter ezubere iche DNA endonuclease (DTECR)].
N'afọ ndị na-adịbeghị anya, ụzọ dị iche iche maka nchọpụta nucleic acids dabeere na CRISPR apụtala [89, 90].Ụzọ dabere na CRISPR omenala na-ewekarị oge ma na-arụsi ọrụ ike n'ihi ọtụtụ usoro gụnyere mmịpụta acid nucleic, mmụba na nchọpụta CRISPR.Ikpughe nke mmiri mmiri na ikuku nwere ike ime ka ohere nke nsonaazụ dị mma dị njọ.Nyere ihe ndị a dị n'elu, usoro ndị dabeere na CRISPR dị oke mkpa nke njikarịcha.
E mepụtala ikpo okwu microfluidic nke na-achịkwa pneumatically nke nwere ike ịrụ nyocha 24 n'otu oge maka ngwa nchọpụta CRISPR-Cas12a na CRISPR-Cas13a [91].Ejiri ngwaọrụ nchọpụta fluorescence na-agafe usoro nucleic acid wee chọpụta DNA femtomolar na RNA na-akpaghị aka.Chen et al.[92] agbakwunyere recombinase amplification na usoro CRISPR-Cas12a na centrifugal microfluidics (Fig. 5a).Ọrụ a na-emeri ihe isi ike nke ijikọ usoro abụọ a n'ihi na Cas12a nwere ike igbari DNA ozi ma gbochie usoro nkwalite ahụ.Na mgbakwunye, Chen et al.[92] echekwabakwara ihe mmeghachi omume reagents na njikwa microfluidic centrifugal iji mechaa usoro niile ozugbo.Na ọrụ ọzọ, Silva et al.[93] mepụtara usoro nyocha na-enweghị CRISPR/Cas12a amplification na smartphone iji chọpụta SARS-CoV-2 (Fig 5b).Nnwale a, nke a maara dị ka sistemụ enweghị nkwalite ekwentị dabere na ekwentị, gụnyere enzyme CRISPR/Cas dabere na nleba anya ama nke akara afụ na-emepụta catalase na ọwa microfluidic.Nchọpụta mmetụta nke ihe na-erughị 50 mbipụta/µl nke nucleic acid na-enweghị nkwalite, usoro dum site na ntụtụ sample ruo n'ịgụ akara na-ewe naanị nkeji 71.
Ụzọ nchọpụta Nucleic acid dabere na CRISPR.Centrifugal POCT maka nchọpụta ihe omimi molecular agbakwunyere dabere na CRISPR (nke sitere na [92]).b Mmepe nke ule CASCADE maka nyocha dabere na smartphone nke SARS-CoV-2 (nke sitere na [93]).RAA recombinase amplification, PAM n'akụkụ protospacer motif, CRISPR kpọkọtara mkpụmkpụ palindromic na-emegharị n'oge oge, usoro CASCADE na-enweghị nkwalite ekwentị na CRISPR / CAS-based enzymes, 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride EDC
Dị ka nzọụkwụ ikpeazụ na nchọpụta nucleic acid, nchọpụta mgbaàmà na-egosipụta kpọmkwem nsonaazụ nchoputa ma bụrụ ihe dị mkpa na mmepe nke POCT dị mma, nke nwere mmetụta na nke ziri ezi.Enwere ike ịgụ akara ngosi site na iji ụzọ dị iche iche dịka fluorescent, electrochemical, colorimetric and magnetization.N'akụkụ a, anyị na-akọwa ihe kpatara usoro nke ọ bụla ma jiri nyocha nke mkpụrụ ndụ nke ọrịa na-efe efe na microfluidics.
A na-eji usoro dabere na fluorescence maka nchọpụta POCT nke ọrịa na-efe efe n'ihi uru dị ịrịba ama nke ezi uche, ọnụ ala dị ala, ịdị mfe nke ọrụ, na nyocha nlekọta nlekọta [94, 95].Atụmatụ ndị a na-eji fluorophores akpọrọ aha dị ka ihe eji esiji fluorescent na nanomaterials mepụta mgbama a na-achọpụta (nkwalite fluorescence ma ọ bụ quenching).Nchoputa a na-egosi na atumatu dabere na fluorescence nwere ike kewaa n'ime akara fluorescent ozugbo, ihe nrịbama, na nchọta fluorescent (96).Nchọpụta akara fluorescent ozugbo na-eji akara fluorescent pụrụ iche iji kpọọ ligands akọwapụtara nke na-emepụta ọnụ ọgụgụ fluorescence akọwapụtara mgbe ahọpụtara ya na ebumnuche.Maka nchọpụta mgbama dabere na fluorescence, ịdị mma nke mgbama fluorescent nwere njikọ chiri anya na oke mmasị.Mmetụta fluorescence bụ nke a na-apụghị ileghara anya na enweghị ebumnuche ma bụrụ nke a na-achọpụta mgbe ọnụọgụ zuru oke dị.N'aka nke ọzọ, ike nke fluorescence achọpụtara site na "signal-off" fluorescence na-adaba na ọnụọgụ nke ebumnuche, na mbụ rute uru kachasị elu ma jiri nwayọọ nwayọọ na-ebelata ka ebumnobi na-abawanye.Dịka ọmụmaatụ, iji CRISPR-Cas13a lekwasịrị anya na-adabere na trans-cleavage usoro, Tian et al.[97] mepụtara atụmatụ njiri mara akwụkwọ akụkọ iji chọpụta RNA ndị na-agafe ntụgharị ederede ozugbo (Fig 6a).Mgbe ejikọtara ya na RNA ndị agbakwunyere, enwere ike ịgbalite mgbagwoju CRISPR-Cas13-RNA, na-ebute mwepu transcollateral site na RNA onye nta akụkọ na-abụghị nke akọwapụtara.A na-emenyụ onye nta akụkọ a na-akpọ fluorescently [fluorophore (F)] site na quencher (Q) na-emebibeghị na fluoresces mgbe mgbagwoju anya na-arụ ọrụ gbawara ya.
Uru nke nchọpụta electrochemical bụ ngwa ngwa nchọpụta dị elu, mmepụta dị mfe, ọnụ ala dị ala, dị mfe iburu na njikwa akpaka.Ọ bụ usoro nyocha siri ike maka ngwa POCT.Dabere na graphene field-mmetụta transistors Gao et al.[98] mepụtara nanobiosensor maka nchọpụta multiplex nke antigens ọrịa Lyme sitere na nje bacteria Borrelia burgdorferi na njedebe nchọpụta nke 2 pg/mL (Fig 6b).
A na-eji nyocha agba agba na ngwa POCT, na-erite uru site na uru dị na mbugharị, ọnụ ala dị ala, ịdị mfe nke nkwadebe, na ọgụgụ anya.Nchọpụta Colorimetric nwere ike iji oxidation nke peroxidase ma ọ bụ peroxidase-dị ka nanomaterials, nchịkọta nke nanomaterials, na mgbakwunye nke ihe e ji esiji ákwà iji gbanwee ozi gbasara ọnụnọ nke nucleic acids lekwasịrị anya na mgbanwe agba a na-ahụ anya [99, 100, 101].N'ụzọ doro anya, a na-eji nanoparticles ọla edo eme ihe n'ọtụtụ ebe na mmepe nke atụmatụ agba agba, na n'ihi ikike ha nwere ime mgbanwe ngwa ngwa na agba agba, enwere mmasị na mmepe nke POCT colorimetric platforms maka na ọnọdụ nyocha nke ọrịa na-efe efe [102].Site na ngwaọrụ centrifugal microfluidic agbakwunyere [103], a na-achọpụta nje ndị na-ebute nri na mmiri ara ehi emetọọ na-akpaghị aka n'ogo nke mkpụrụ ndụ nje iri, na enwere ike ịgụpụta nsonaazụ ya n'ime nkeji 65 (Fig 6c).
Usoro nhụta magnetik nwere ike ịchọpụta nyocha nke ọma site na iji ihe ndọta, ma enweela mmasị dị ukwuu na ngwa POCT n'ime iri afọ ndị na-adịbeghị anya.Usoro nhụta magnetik nwere ụfọdụ uru pụrụ iche dị ka ihe ndọta dị ọnụ ala karịa ngwa anya ngwa anya dị oke ọnụ.Otú ọ dị, iji oghere magnetik na-eme ka nchọpụta nchọpụta dịkwuo mma ma na-ebelata oge nkwadebe sample [104].Tụkwasị na nke ahụ, nsonaazụ nke nyocha magnetik na-egosipụta nkọwa dị elu, mmetụta uche, na mgbama dị elu na ụda mkpọtụ n'ihi nrịbama ndọta na-abaghị uru nke ihe atụ ndụ [105].Sharma et al.agbakwunyere njikọ biosensor nke ọwara ndọta n'ime ikpo okwu microchip enwere ike.[106] maka nchọpụta multiplex nke ọrịa nje (Fig 6d).Biosensor ji nlezianya na-achọpụta acid nucleic subnanomolar dịpụrụ adịpụ na nje nje.
Usoro nchọpụta mgbaama a na-ahụkarị.Echiche nke nchọpụta hyperlocalized nke Cas13a (nke sitere na [97]).b Graphene nanobiosensor FET jikọtara ya na Lyme GroES scFv (nke sitere na [98]).c Ihe ngosi Colorimetric maka nchọpụta multiplex nke nri na-eri nri na mgbawa centrifugal microfluidic: Nke 1 na nke 3 ihe atụ nwere ihe ndị na-ebute ụzọ, na nke 2, No. 4 na No. 5 samples na-enweghị ebumnuche ebumnuche (nke sitere na [103]) .d Biosensor dabere na njikọ ọwara magnetik, gụnyere ikpo okwu, ihe nkwuchita nke arụnyere arụnyere, ngalaba njikwa, na ihe na-enye ike maka ọgbọ mgbaàmà / nweta (nke sitere na [106]).GFET Graphene FET, Escherichia coli, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus, Vibrio parahaemolyticus, Listeria monocytogenes, PC PC, PDMS Dimethicone, PMMA polymethyl methacrylate.
N'agbanyeghị njirimara mara mma nke ụzọ nchọpụta ndị dị n'elu, ha ka nwere adịghị ike.A na-atụnyere usoro ndị a (tebụl 1), gụnyere ụfọdụ ngwa nwere nkọwa (uru na ọghọm).
Site na mmepe nke microfluidics, sistemụ microelectromechanical, nanotechnology na sayensị ihe, iji microfluidic ibe maka ịchọpụta ọrịa na-efe efe na-aga n'ihu mgbe niile [55,96,107,108].Ntuzi aka nke ọma nke obere akụrụngwa na mmiri mmiri na-enye aka n'ịchọpụta izi ezi na ọnụ ahịa ọnụ ahịa.Ya mere, maka mmepe n'ihu, agbaala mbọ iji kwalite na kwalite ibe, na-ebute ibe microfluidic dị iche iche nwere nhazi na ọrụ dị iche iche.N'ebe a, anyị na-ewebata nkenke ụdị dị iche iche nke nyiwe microfluidic a na-ahụkarị ma tụle njirimara ha (uru na ọghọm).Na mgbakwunye, ọtụtụ n'ime ihe atụ ndị edepụtara n'okpuru na-elekwasị anya na mbuso agha SARS-CoV-2.
LOCC bụ sistemụ nyocha dị mgbagwoju anya nke pere mpe na arụ ọrụ ha dị ntakịrị, agbakwunyere, akpaghị aka na nke yiri ya site na ntụtụ na nkwadebe, njikwa mmiri na nchọpụta mmiri [109,110].A na-emegharị mmiri mmiri site na geometry ejiri nlezianya hazie yana mmekọrịta nke ọtụtụ mmetụta anụ ahụ dị ka gradients nrụgide, capillary action, electrodynamics, field magnet and acoustic ebili mmiri [111].LOCC na-egosi uru dị mma na nyocha dị elu na nchọpụta ọtụtụ, na ngwa ngwa nyocha ngwa ngwa, obere ihe nlele, obere ike oriri, na njikwa dị elu na arụmọrụ arụmọrụ;Otú ọ dị, ngwaọrụ LOCC dị nnọọ nro, na imepụta ihe, nkwakọ ngwaahịa, na ntinye.Otú ọ dị, imegharị ihe na imegharị ihe na-eche nnukwu nsogbu ihu [96].Tụnyere nyiwe ndị ọzọ, LOCC nwere uru pụrụ iche n'ihe gbasara ụdị ngwa dị iche iche yana ndakọrịta teknụzụ kacha mma, mana ọghọm ya pụtakwara ìhè, ya bụ mgbagwoju anya dị elu na enweghị ike ikwughachi.Ndabere na nfuli dị n'èzí, nke na-adịkarị oke ọnụ ma dị oke ọnụ, na-egbochikwa ojiji ha na POCT.
N'oge ntiwapụ COVID-19, LOCC nwetara nlebara anya dị ukwuu.N'otu oge ahụ, enwere ọtụtụ ibe ọhụrụ na-ejikọta ọtụtụ teknụzụ.Dịka ọmụmaatụ, a na-eji smartphones eme ihe ugbu a dị ka ngwaọrụ nyocha enwere ike ibugharị ma nwee nnukwu ikike maka ntinye LOCC.Sun et al.[21] chepụtara mgbawa microfluidic nke na-enye ohere ịgbatị usoro nucleic acid kpọmkwem nke nje ise, gụnyere SARS-CoV-2, na-eji LAMP wee nyochaa ha site na iji ekwentị n'ime elekere 1 ka njedebe nke mmeghachi omume ahụ gasịrị.Dị ka ihe atụ ọzọ, Sundah et al.[112] mepụtara mgba ọkụ molecular [catalytic amplification by molecular transition state switch (CATCH)] maka nchọpụta ozugbo na nke nwere mmetụta nke SARS-CoV-2 RNA na-eji ekwentị. CATCH dakọtara na LOCC nwere ike ibugharị ma nweta arụmọrụ dị elu (ihe dịka 8 RNA mbipụta/μl; <1 h na ọnụ ụlọ okpomọkụ) [112]. CATCH dakọtara na LOCC nwere ike ibugharị ma nweta arụmọrụ dị elu (ihe dịka 8 RNA mbipụta/μl; <1 h na ọnụ ụlọ okpomọkụ) [112]. CATCH совместим с портативным LOCC и обеспечивает превосходную производительность ( примерно 8 копий Рентерно 8 копий Реть . CATCH dakọtara na LOCC nwere ike ibugharị ma na-enye mmepụta dị mma (ihe dịka 8 RNA ụdị/µl; <1 h na ọnụ ụlọ okpomọkụ) [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 производной производительностью ( примерно 8 копий Рим . CATCH dakọtara na LOCC ndị a na-ebugharị ebugharị ma nwee ọmarịcha arụmọrụ (ihe dịka 8 RNA nnomi/µl; <1 hour na ụlọ okpomọkụ) [112].Na mgbakwunye, ngwaọrụ LOCC maka nyocha mkpụrụ ndụ na-ejikwa ụfọdụ ike ịnya ụgbọ ala dịka oghere, ịgbatị, na oghere eletrik.Kang et al.[113] gosipụtara ezigbo oge, ultra-fast nanoplasma-on-a-chip PCR maka nyocha ngwa ngwa na ọnụọgụ nke COVID-19 n'ọhịa site na iji mgbawa PCR vacuum plasmonic liquid.Li et al.[114] mechara mepụta mgbawa microfluidic gbatịpụrụ na-enyere aka ịchọpụta COVID-19.Ikpo okwu na-eji sistemụ nkwalite RT-LAMP iji chọpụta ma nlele ọ dị mma ma ọ bụ adịghị mma.N'ikpeazụ, Ramachandran et al.[115] nwetara gradients ọkụ eletrik kwesịrị ekwesị site na iji isotachophoresis (ITP), usoro nlebara anya ion nke etinyere na microfluidics.Site na ITP, RNA lekwasịrị anya sitere na sample nasopharyngeal swab raw nwere ike sachaa ozugbo.Mgbe ahụ Ramachandran et al.[115] Ijikọta nchacha ITP a na LAMP na-akwalite ITP na nyocha CRISPR achọpụtara SARS-CoV-2 na swab nasopharyngeal mmadụ na ụdị ụlọ ọgwụ n'ihe dị ka nkeji iri atọ na ise.Na mgbakwunye, echiche ọhụrụ na-apụta mgbe niile.Jadhav et al.[116] tụpụtara atụmatụ nchoputa nke gbadoro ụkwụ n'elu Raman spectroscopy jikọtara ya na ngwaọrụ microfluidic nwere ma carbon nanotubes ọla edo nwere mkpuchi kwụ ọtọ kwụ ọtọ ma ọ bụ eletrọspun micro/nanotubes nwere ike ịtụfu.Igwe igwe nzacha arụrụ arụrụ arụrụ arụrụ arụrụ n'ime membrane ka enwere ike ịtụfu ya.Ngwa ahụ na-eme ka nje virus si na mmiri dị iche iche dị iche iche dị ka mmiri, nasopharynx na anya mmiri.Ya mere, ọnụọgụ nje dị ukwuu ma enwere ike mata nje a nke ọma site na mbinye aka Raman.
LOAD bụ centrifugal microfluidic ikpo okwu nke a na-achịkwa usoro niile site na usoro ugboro ugboro nke na-atụgharị mkpụrụ nke microstructured [110].A na-eji ngwaọrụ LOAD mara ya site n'iji ike centrifugal dị ka ike ịkwọ ụgbọala dị mkpa.Mmiri mmiri na-anọkwa n'okpuru capillary, ndị agha Euler na Coriolis.N'iji ngwaọrụ centrifuge, a na-eme nyocha na mmiri mmiri na-aga n'ihu na-arụ ọrụ site na radial n'ime ruo n'èzí, na-ewepụ mkpa maka tubing mpụga ọzọ, nfuli, actuators, na valves na-arụ ọrụ.Na nkenke, otu usoro njikwa na-eme ka ọrụ dị mfe.Ndị agha na-arụ ọrụ na mmiri mmiri na otu microfluidic ọwa n'otu ebe dị anya site na ibu ibu bụ hà nhata, nke na-eme ka o kwe omume ịmegharị usoro ọwa ahụ.Ya mere, ngwa LOAD dị mfe ma dị oke ọnụ ahịa iji chepụta na imepụta karịa ngwa LOCC nkịtị, ebe mmeghachi omume na-adabere na ya na nke yiri ya;Otú ọ dị, n'ihi ike ọrụ dị elu nke akụrụngwa centrifugal, ihe mgbawa dị adị nwere oke na obere mpịakọta siri ike.na ụgbọ ala.N'otu oge ahụ, a na-emepụta ọtụtụ ngwaọrụ LOAD maka naanị otu, nke dị oke ọnụ maka nchọpụta buru ibu [96, 117, 118, 119].
N'ime iri afọ ndị na-adịbeghị anya, LOAD, nke a na-ewere dị ka otu n'ime ngwaọrụ microfluidic kachasị mma, enwetala nlebara anya dị ukwuu site n'aka ndị nchọpụta na ndị nrụpụta.Ya mere, LOAD enwetala nnabata zuru oke ma ejiri ya mee ihe maka nchọpụta mkpụrụ ndụ nke nje na-efe efe [120, 121, 122, 123, 124], ọkachasị n'oge ntiwapụ COVID-19.Dịka ọmụmaatụ, na njedebe nke 2020, Ji et al.[60] gosiputara nyocha RT-qPCR ozugbo maka nchọpụta ngwa ngwa na akpaaka nke SARS-CoV-2 yana ọrịa influenza A na B na akpịrị swab.Mgbe ahụ Xiong et al.[74] gosipụtara ikpo okwu discoid microfluidic jikọtara LAMP maka nchọpụta ngwa ngwa, nke ziri ezi na n'otu oge nke coronaviruses iku ume mmadụ asaa, gụnyere SARS-CoV-2, n'ime nkeji iri anọ.Na mbido 2021, de Oliveira et al.[73] gosiputara mgbawa microfluidic polystyrene toner centrifugal, ejiri aka na-eji mkpịsị aka arụ ọrụ, maka nyocha mkpụrụ ndụ RT-LAMP nke COVID-19.N'ikpeazụ, Dignan et al.[39] gosipụtara microdevice centrifuge obere akpaaka maka ịsacha SARS-CoV-2 RNA ozugbo site na ngalaba swab.Medved et al.[53] tụpụtara usoro inline SARS-CoV-2 aerosol mgbawa nwere obere mgbawa microfluidic fluorescent mgbawa nwere oke nchọpụta nke mbipụta 10 / μL yana opekempe okirikiri nke nkeji iri na ise.Suarez et al.[75] kọrọ na nso nso a mmepe nke igwe okwu centrifugal microfluidic modular agbakwunyere maka nchọpụta kpọmkwem nke SARS-CoV-2 RNA na ncha ọkụ nasopharyngeal swab na-ekpo ọkụ na-eji LAMP.Ihe atụ ndị a na-egosi nnukwu uru na nkwa nke LOAD na nyocha mkpụrụ ndụ nke COVID-19.
Na 1945 Muller na Clegg [125] bu ụzọ wepụta ọwa microfluidic na mpempe akwụkwọ site na iji akwụkwọ nzacha na paraffin.Na 2007, ndị otu Whitesides [126] mepụtara ikpo okwu akwụkwọ mbụ na-arụ ọrụ maka nnwale protein na glucose.Akwụkwọ abụrụla ihe dị mma maka microfluidics.Akwụkwọ ahụ nwere ihe ndị dị n'ime ya dị ka hydrophilicity na porous Ọdịdị, ọmarịcha biocompatibility, ịdị arọ dị arọ, mgbanwe, mpịakọta, ọnụ ala dị ala, ịdị mfe iji na ịdị mma.µPAD oge gboo nwere ihe owuwu hydrophilic/hydrophobic wuru na mpempe akwụkwọ.Dabere na nhazi akụkụ atọ, μPAD nwere ike kewaa n'ime akụkụ abụọ (2D) na akụkụ atọ (3D) μPAD.A na-emepụta 2D µPAD site n'ịme oke hydrophobic iji mepụta ọwa microfluidic, ebe 3D µPAD na-emekarị site na nchịkọta nke mpempe akwụkwọ microfluidic 2D, mgbe ụfọdụ site na mpịachi akwụkwọ, usoro slip, oghere oghere, na mbipụta 3D [96].A na-achịkwa mmiri mmiri ma ọ bụ ihe ndị dị ndụ na μPAD site na ike capillary na-enweghị isi iyi ike dị na mpụga, na-eme ka nchekwa nke reagents, njikwa ihe nlele, na nchọpụta multiplex.Otú ọ dị, njikwa ọsọ ziri ezi na nchọpụta multiplex na-egbochi site na ezughị ezu nchọta ọsọ ọsọ, uche, na reusability [96, 127, 128, 129, 130].
Dị ka ikpo okwu microfluidic na-adịghị ahụkebe, μPAD akwalitela ma mepụta ya maka nyocha mkpụrụ ndụ nke ọrịa na-efe efe dị ka HCV, HIV, na SARS-CoV-2 [131, 132].Maka nchọpụta nhọrọ na nke nwere mmetụta nke HCV, Tengam et al.[133] mepụtara biosensor akwụkwọ akụkọ dabere na akwụkwọ fluorescent na-eji nyocha nucleic acid nke ukwuu dabere na pyrrolidinyl peptide.Nucleic acid bụ covalently immobilized na akụkụ oxidized cellulose akwụkwọ site reductive alkylation n'etiti amino otu na aldehyde otu, na nchọpụta dabeere fluorescence.Enwere ike ịgụ akara ngosi ndị a site na ngwa emebere nke ọma yana igwefoto fluorescent nwere ike ijikọ yana igwefoto ekwentị.N'ikpeazụ, Lu et al.[134] haziri electrode na-agbanwe agbanwe dabere na nickel/gold nanoparticles/carbon nanotubes/polyvinyl alcohol organometallic framework composites maka nchọpụta nje HIV site na hybridization DNA site na iji methylene blue dị ka ihe ngosi DNA redox.N'oge na-adịbeghị anya, Chowdury et al.[135] weputara ihe nlere anya n'elu ikpo okwu maka nnwale µPAD nlebara anya site na iji amiri onye ọrịa na ngwakọta LAMP na teknụzụ onyonyo obere maka nchọpụta nyocha COVID-19.
Nnwale nrịba dị n'akụkụ na-eduzi mmiri mmiri site na ike capillary yana ịchịkwa mmegharị mmiri site na wettability na njirimara nke ihe ndị nwere oghere ma ọ bụ obere ihe.Ngwaọrụ eruba n'akụkụ nwere ihe nlele, conjugate, incubator na nchọpụta, na paịlị na-amịkọrọ ihe.Ụmụ irighiri ihe nucleic acid dị na LFA na-amata ihe nkedo akọwapụtara nke edobere na saịtị njikọ wee kee dị ka ogige.Ka mmiri mmiri na-agafe na mpempe akwụkwọ ntinye na nchọpụta, a na-ejide ogige ndị ahụ site na mkpụrụ ndụ ihe nkedo nke dị na ule na akara akara, na-egosi nsonaazụ ndị nwere ike ịgụ ozugbo na anya gba ọtọ.Dịka, LFA nwere ike mechaa na nkeji 2-15, nke dị ngwa karịa nchọpụta ọdịnala.N'ihi usoro pụrụ iche, LFA chọrọ ọrụ ole na ole ma ọ dịghị achọ ngwa mgbakwunye, nke na-eme ka ọ bụrụ enyi na enyi.Ọ dị mfe imepụta na miniaturize, na ọnụ ahịa nke mpempe akwụkwọ dabere na ya dị ala.Otú ọ dị, a na-eji ya naanị maka nyocha nke qualitative, na nchọpụta ọnụọgụ dị oke ike, na ikike multiplexing na mmepụta dị oke oke, na naanị otu nucleic acid zuru ezu nwere ike ịchọta n'otu oge [96,110,127].
Ọ bụ ezie na ọtụtụ ngwa nke LFA na-elekwasị anya na immunoassays, iji LFA maka nchọpụta ihe omimi na ibe microfluidic dịkwa irè na nke a ma ama [136].N'ihe banyere nje ịba ọcha n'anya B, HIV na SARS-CoV-2 LFA Gong et al.[137] tụpụtara usoro ikpo okwu nanoparticle LFA na-agbanwe agbanwe wee gosipụta ịdị iche iche nke ikpo okwu dị ntakịrị na nke enwere ike ibugharị site na nchọta nwere mmetụta na ọnụọgụ nke ọtụtụ ebumnuche dịka HBV nucleic acid.Na mgbakwunye, Fu et al.[138] gosipụtara akwụkwọ akụkọ LFA nke dabere na Raman spectroscopy nke elu maka nyocha ọnụọgụ nke HIV-1 DNA na obere mkpokọta.Maka nchọpụta ngwa ngwa na nke siri ike nke SARS-CoV-2, Liu et al.[85] mepụtara microfluidic-integrated RPA lateral flow analysis site na ijikọta RT-RPA na usoro nchọpụta n'akụkụ ụwa n'ime otu usoro microfluidic.
Ngwa nke dị iche iche microfluidic nyiwe dịgasị iche dabere na kpọmkwem ọmụmụ, na-eji zuru oke nke ike na uru nke nyiwe.Site na valvụ dị ọnụ ala, nfuli na ducts, LOCC bụ ikpo okwu zuru oke maka ụdị ngwa dị iche iche na mmekorita ya na ọnụ ụlọ kachasị maka mmepe.Ya mere, anyị na-atụ anya ma na-akwado na a ga-eme ọmụmụ ihe ọhụrụ na LOCC dị ka mgbalị mbụ na ka emeziwanye ọnọdụ ahụ.Tụkwasị na nke ahụ, a na-atụ anya ka a chọpụta ụzọ dị irè na nke ziri ezi ma jiri ya mee ihe na usoro ahụ.LOAD karịrị n'ụzọ ziri ezi njikwa mmiri sitere na ngwaọrụ LOCC dị ma gosipụta uru pụrụ iche na draịva otu site na ike centrifugal na-enweghị mkpa draịva mpụga, ebe nzaghachi ndị yiri ya nwere ike kewapụ ma mekọrịta.Ya mere, n'ọdịniihu, LOAD ga-abụ isi okwu microfluidic na-arụ ọrụ obere aka yana teknụzụ tozuru oke na akpaaka.Igwe okwu µPAD na-ejikọta uru nke LOCC na ihe ndị dabeere na akwụkwọ maka ọnụ ala dị ọnụ ala, nchọpụta otu ojiji.Ya mere, mmepe n'ọdịnihu kwesịrị ilekwasị anya na teknụzụ dị mma na nke ọma.Na mgbakwunye, LFA dabara nke ọma maka nchọpụta anya gba ọtọ, na-ekwe nkwa ibelata oriri sample na ngwa ngwa nchọpụta.E gosipụtara ntụnyere ikpo okwu zuru ezu na Tebụl 2.
Nyocha dijitalụ na-ekewa ihe nlele ahụ n'ime ọtụtụ microreactors, nke ọ bụla n'ime ha nwere ọnụọgụ pụrụ iche nke ụmụ irighiri ihe dị iche iche [139, 140].Nnwale dijitalụ na-enye uru dị ukwuu maka ịrụ ọnụọgụgụ zuru oke site n'ịme puku kwuru puku nnwale biochemical n'otu oge na n'otu n'otu na ngalaba ọnụ ọgụgụ micron karịa na usoro na-aga n'ihu.E jiri ya tụnyere microfluidics ọdịnala, mmeghachi omume nke ngalaba nwere ike ibelata olu nlele, ịbawanye arụmọrụ mmeghachi omume, wee jikọta ya na usoro nyocha ndị ọzọ na-enweghị mkpa ọwa, nfuli, valves na atụmatụ kọmpat [141, 142, 143, 144, 145, 146. 147].A na-eji ụzọ abụọ na-esonụ na nyocha dijitalụ iji nweta otu na nkewa ziri ezi nke ngwọta, gụnyere reagents na ihe atụ dị ka sel, nucleic acid, na ihe ndị ọzọ ma ọ bụ molecules: (1) dobe emulsions na-erigbu enweghị ntụkwasị obi nke mmiri mmiri;(2) A na-eme nkewa n'usoro site na mgbochi geometric nke ngwaọrụ ahụ.Na usoro nke mbụ, enwere ike ịmepụta ụmụ irighiri mmiri nwere reagents na ihe nlele na microchannels site na ụzọ ndị na-agafe agafe dị ka co-current, crossflow, flow focus, emulsification, microchannel emulsification, na membranes site na viscous shear Forces and emulsification with channel change.localization [143, 145, 146, 148, 149] ma ọ bụ iji ụzọ arụ ọrụ [150, 151], nke na-ewebata ike ọzọ site na njikwa eletriki, magnetik, thermal na n'ibu.Na usoro nke ikpeazụ, a na-ekerịta nha nha nha kachasị mma n'ime ụlọ microfluidic site na idobe ihe owuwu oghere nke otu nha, dị ka micropits na arrays elu [152,153,154].N'ụzọ doro anya, ụmụ irighiri mmiri bụ akụkụ dị mkpa nke na-asọpụta nke nwekwara ike ịmepụta ma jiri ya mee ihe n'usoro electrode dabere na microfluidics dijitalụ (DMF).Electrowetting nke dielectrics bụ otu n'ime echiche DMF kacha mma amụrụ, ebe ọ bụ na electrowetting nke dielectrics na-enye ohere ịmegharị nke ọma nke onye ọ bụla, na-achịkwa ọdịdị nke mmiri mmiri na akara eletrik asymmetric na-agafe n'akụkụ dị iche iche [141, 144].Ọrụ bụ isi na ụmụ irighiri mmiri dị na DMF gụnyere nhazi, kewaa, na ijikọ [151, 155, 156], nke enwere ike itinye n'ọrụ n'akụkụ dị iche iche nke nyocha, karịsịa na nchọpụta molecular [157, 158, 159].
Nchọpụta nucleic acid dijitalụ bụ teknụzụ nyocha mkpụrụ ndụ nke ọgbọ nke atọ na-eso PCR ọdịnala na PCR nke adịgboroja (qPCR), n'otu aka ahụ yana usoro ntinye dị elu yana biopsy mmiri mmiri.N'ime afọ iri abụọ gara aga, acid nucleic dijitalụ etolitela ngwa ngwa n'ọhịa nke nchọpụta ihe omimi nke nje na-efe efe [160, 161, 162].Nchọpụta zuru oke nke nchọpụta nucleic acid dijitalụ na-amalite site n'ịkwakọba ihe nlele na reagents n'ime ụlọ nke ọ bụla iji hụ na usoro ebumnuche ọ bụla nwere otu ohere ịbanye n'ime ụlọ ọ bụla.N'usoro iwu, enwere ike kenye ngalaba ọ bụla ọtụtụ usoro ebumnuche, ma ọ bụ enweghị ike ịnwe sistemụ microreaction nọọrọ onwe ya.Site na usoro nhụta dị iche iche akọwara n'elu, akụkụ nwere usoro ebumnuche microbial nke na-emepụta akara n'elu otu ụzọ nwere ike iji anya gba ọtọ ma ọ bụ igwe jiri anya ya wee kpọọ ya dị ka ihe dị mma, ebe a na-akpọ akụkụ ndị ọzọ na-emepụta akara n'okpuru ọnụ ụzọ dị ka ihe dị mma. .ndị na-adịghị mma, nke na-eme ka mgbaàmà maka ngalaba ọ bụla bụrụ boolean.Ya mere, site n'ịgbakọ ọnụ ọgụgụ nke ngalaba ndị e kere na ọnụ ọgụgụ nke nsonaazụ dị mma mgbe mmeghachi omume ahụ gasịrị, a ga-ejikọta akwụkwọ mbụ nke ihe nlele ule site na iji usoro nkesa Poisson na-enweghị mkpa maka ọkọlọtọ ọkọlọtọ, nke a chọrọ maka nyocha ọnụọgụgụ oge niile dị otú ahụ. dị ka qPCR.[163] E jiri ya tụnyere usoro nchọpụta mkpụrụ ndụ ọdịnala, nchọpụta nucleic acid dijitalụ nwere ogo dị elu nke akpaaka, ọsọ nyocha dị elu na nghọta, obere reagents, obere mmetọ, yana nhazi na nrụpụta dị mfe.Maka ebumnuche ndị a, ojiji nke nyocha dijitalụ, ọkachasị ụzọ dabere, maka nyocha mkpụrụ ndụ, ijikọ mmụba na usoro ịgụta akara, amụrụla nke ọma n'oge ntiwapụ nke SARS-CoV-2.Dịka ọmụmaatụ, Yin et al.[164] ụzọ PCR dijitalụ jikọtara ọnụ na ngwa ngwa iji chọpụta mkpụrụ ndụ ihe nketa ORF1ab, N na RNase P na SARS-CoV-2 na mgbawa microfluidic.N'ụzọ doro anya, usoro ahụ nwere ike ịchọpụta mgbaàmà dị mma n'ime 115 sekọnd, nke dị ngwa ngwa karịa PCR nkịtị, na-egosi ịdị irè ya na nchọpụta nleba anya (Nyocha 7a).Dong et al.[165], Sow et al.[157], Chen et al.[166] na Alteri et al.[167] tinyekwara PCR dijitalụ droplet (ddPCR) iji chọpụta SARS-CoV-2 na sistemụ microfluidic nwere nsonaazụ dị egwu.Iji meziwanye ọnụego nchọpụta ahụ, Shen et al.[168] nwetara onyonyo mgbawa dabere na ddPCR n'ihe dị ka 15 s na-ejighi usoro ịnya ihe onyonyo, na-eme ka usoro teknụzụ ddPCR dị ngwa site na ụlọ nyocha gaa na ngwa.Ọ bụghị naanị ụzọ nkwalite okpomọkụ dị ka PCR ka a na-etinye, mana a na-ejikwa ụzọ nkwalite isothermal mee ka ọnọdụ mmeghachi omume dị mfe na nzaghachi ngwa ngwa.Lu et al.[71] mepụtara SlipChip maka nyocha droplet, nke nwere ike iwepụta ụmụ irighiri mmiri dị iche iche na njupụta dị elu n'otu nzọụkwụ yana ọnụọgụ SARS-CoV-2 nucleic acid site na iji LAMP dijitalụ (Nyocha 7b).Dị ka nkà na ụzụ na-agbanwe ngwa ngwa, CRISPR nwekwara ike ịrụ ọrụ dị mkpa na nchọpụta nucleic acid dijitalụ site na imaging colorimetric dị mma na-enweghị mkpa ọzọ nucleic acid stains.Ackerman et al.mepụtara mmeghachi omume matrix combinatorial maka nyocha multiplex nke nucleic acid.[158] achọpụtara nje 169 metụtara mmadụ, gụnyere SARS-CoV-2, n'ime ụmụ irighiri mmiri nwere CRISPR-Cas13 nucleic acid reagents reagents na microwell assay (Nyocha 7c).Tụkwasị na nke ahụ, a pụrụ iji nkwalite isothermal na nkà na ụzụ CRISPR mee ihe n'otu usoro iji jikọta uru nke abụọ ahụ.Park et al.[169] Emebere nyocha dijitalụ CRISPR/Cas12a n'ime mgbawa microfluidic azụmahịa maka nchọpụta SARS-CoV-2 amịpụtara na ọkụ gburu dabere na otu ọkwa RT-RPA nwere njiri dị mkpụmkpụ na nke dị elu na nchọpụta ndabere. oge ruru., obosara ike nso na mma uche (Fig. 7d).Enyere nkọwa ụfọdụ nke ihe atụ ndị a na Tebụl 3.
Igwe okwu dijitalụ a na-ahụkarị maka nchọpụta nucleic acid.a Usoro PCR dijitalụ ngwa ngwa nwere usoro isi anọ: nkwadebe nlele, nkesa ngwakọta mmeghachi omume, usoro mmụba, na nha nha (nke sitere na [164]).b Schematic na-egosi nyocha nke ụmụ irighiri mmiri SlipChip maka nhazi droplet na njupụta dị elu (nke sitere na [71]).c CARMEN-Cas workflow eserese13 (nke sitere na [158]).d Nyochaa nchọpụta nje dijitalụ dị elu yana CRISPR/Cas n'otu ite (emetere na [169]).W/O mmiri na mmanụ, polydimethylsiloxane PDMS, PCR polymerase chain reaction, DAQ data collection, PID proportional integral derivative, CARMEN combinatorial matrix mmeghachi omume maka multiplex nucleic acid nwale, SARS-CoV-2, oké iku ume ọrịa, coronavirus 2 , Mmụba RT nke reverse transcriptase recombinase polymerase-RPA, akara S/B n'azụ.