Ua tsaug rau koj tuaj xyuas www.chinacdoe.com.Qhov browser version koj siv tau txwv CSS kev txhawb nqa.Rau qhov kev paub zoo tshaj plaws, peb xav kom koj siv qhov browser tshiab (lossis lov tes taw Compatibility Hom hauv Internet Explorer).Nyob rau lub sijhawm no, txhawm rau ua kom muaj kev txhawb nqa txuas ntxiv, peb yuav ua rau lub xaib tsis muaj qauv thiab JavaScript.
Lab-on-a-chip systems nrog lub peev xwm ntawm qhov chaw muab lub peev xwm rau kev kuaj mob sai thiab raug thiab muaj txiaj ntsig zoo hauv cov chaw muaj kev txwv tsis pub siv cov khoom siv biomedical thiab cov kws tshaj lij tsis muaj.Txawm li cas los xij, tsim kom muaj qhov ntsuas qhov ntsuas ntawm kev saib xyuas ib txhij muaj tag nrho cov yam ntxwv tsim nyog rau kev siv ntau yam, kev tso tawm raws li qhov xav tau, kev ua haujlwm tau zoo, thiab kev cia ntev ntev ntawm cov tshuaj reagents tseem yog ib qho kev sib tw loj.Ntawm no peb piav qhia txog lub tshuab ua haujlwm micro mus ncig ua si uas tuaj yeem tswj cov kua dej hauv txhua qhov kev taw qhia, muab cov lus teb meej thiab sib npaug rau kev siv cua siab, thiab nyob ruaj khov tiv thaiv tam sim ntawd txav thiab vibrations.Raws li cov thev naus laus zis, peb kuj piav qhia txog kev txhim kho ntawm polymerase saw cov tshuaj tiv thaiv kab mob uas sib koom ua ke reagent qhia, sib xyaw thiab cov tshuaj tiv thaiv ua haujlwm tag nrho hauv ib txheej txheem, uas ua tiav "cov qauv-hauv-lus-tawm" kev ua tau zoo rau txhua qhov kev kuaj mob qhov ntswg los ntawm 18 tus neeg mob. Influenza thiab 18 tus neeg tswj hwm, nyob rau hauv kev sib raug zoo ntawm fluorescence siv nrog tus qauv polymerase saw cov tshuaj tiv thaiv (Pearson coefficients> 0.9).Raws li cov thev naus laus zis, peb kuj piav qhia txog kev txhim kho ntawm polymerase saw cov tshuaj tiv thaiv kab mob uas sib koom ua ke reagent qhia, sib xyaw thiab cov tshuaj tiv thaiv ua haujlwm tag nrho hauv ib txheej txheem, uas ua tiav "cov qauv-hauv-lus-tawm" kev ua haujlwm rau txhua qhov kev kuaj mob qhov ntswg los ntawm 18 tus neeg mob. Nrog Influenza thiab 18 tus neeg tswj hwm, nyob rau hauv kev sib raug zoo ntawm fluorescence siv nrog tus qauv polymerase saw cov tshuaj tiv thaiv (Pearson coefficients> 0.9).Основываясь на этой технологии, мы также описываем разработку системы полимеразной цепной ренквет, ведения реагентов, смешивания и реакции в одном процессе, что обеспечивает выполнение «обрахец-в-ссе» образцов из носа от 18 пациентов с Грипп thiab 18 отдельных контролей, в хорошем соответствии интенсивности флуоресценции стандартной полим ффициенты Пирсона> 0,9).Raws li cov thev naus laus zis no, peb kuj piav qhia txog kev txhim kho cov kab ke polymerase saw tshuaj tiv thaiv uas sib xyaw ua haujlwm ntawm kev txhaj tshuaj, sib tov, thiab ua haujlwm hauv ib txheej txheem, ua rau cov qauv-hauv- teb-tawm rau txhua qhov chaw kho qhov ntswg los ntawm 18 tus neeg mob khaub thuas.thiab 18 tus neeg tswj hwm, hauv kev pom zoo nrog tus qauv polymerase saw tshuaj tiv thaiv fluorescence siv (Pearson's coefficients> 0.9).Raws li cov thev naus laus zis no, peb kuj piav qhia txog kev txhim kho ntawm polymerase saw cov tshuaj tiv thaiv kab mob uas sib xyaw ua ke reagent txhaj, sib tov, thiab cov tshuaj tiv thaiv kev ua haujlwm los tshuaj xyuas txhua qhov chaw kho qhov ntswg los ntawm 18 tus qauv ntawm tus neeg mob qhov ntswg.Influenza thiab 18 tus neeg tswj hwm, fluorescence sib tw. zoo nrog cov txheej txheem polymerase saw cov tshuaj tiv thaiv (Pearson tus coefficient> 0.9).Lub platform tau lees paub kev ntseeg siab ntawm kev tshuaj ntsuam xyuas biomedical thiab yog li tuaj yeem ua kom nrawm rau kev lag luam ntawm ntau yam ntawm cov cuab yeej ntsuas kev saib xyuas.
Cov kab mob tshwm sim hauv tib neeg, xws li 2020 COVID-19 kis thoob qhov txhia chaw uas tau ua rau ntau lab tus tib neeg lub neej, ua rau muaj kev hem thawj loj rau ntiaj teb kev noj qab haus huv thiab tib neeg kev vam meej1.Thaum ntxov, kev kuaj pom cov kab mob sai thiab raug yog qhov tseem ceeb los tswj kev kis tus kab mob thiab txhim kho cov txiaj ntsig kev kho mob.Lub ecosystem kev kuaj mob tseem ceeb raws li lub hauv paus chaw kuaj mob uas kuaj cov qauv xa mus rau tsev kho mob lossis chaw kuaj mob kuaj mob thiab khiav los ntawm cov kws tshaj lij tam sim no txwv kev nkag mus rau ze li 5.8 billion tus neeg thoob ntiaj teb, tshwj xeeb tshaj yog cov neeg nyob hauv cov chaw muaj kev txwv.qhov twg tsis muaj cov khoom siv biomedical kim thiab cov kws tshaj lij uas tsim nyog.Cov kws kho mob 2. Yog li, yuav tsum tau tsim kho qhov pheej yig thiab tus neeg siv-phooj ywg lab-on-a-chip system nrog lub peev xwm ntawm qhov chaw kuaj mob (POCT) uas tuaj yeem muab cov kws kho mob nrog cov ntaub ntawv kuaj mob raws sij hawm los txiav txim siab kuaj mob. .thiab kev kho mob 3.
Lub Koom Haum Ntiaj Teb Kev Noj Qab Haus Huv (WHO) cov lus qhia hais tias POCT zoo tshaj plaws yuav tsum muaj tus nqi pheej yig, siv tau yooj yim (yooj yim siv nrog kev cob qhia tsawg), raug (tsis txhob muaj qhov tsis zoo lossis qhov tsis zoo), nrawm thiab txhim khu kev qha (muab cov khoom zoo rov ua dua), thiab khoom xa tuaj (muaj peev xwm khaws cia ntev thiab yooj yim rau cov neeg siv kawg) 4.Yuav kom ua tau raws li cov kev cai no, POCT systems yuav tsum muab cov yam ntxwv hauv qab no: kev siv ntau yam kom txo tau cov kev cuam tshuam ntawm phau ntawv, kev xav tau tso tawm kom ntsuas cov reagent thauj mus rau qhov ntsuas tau raug, thiab txhim khu kev qha ua haujlwm kom tiv taus ib puag ncig kev co.Tam sim no, cov cuab yeej siv POCT feem ntau yog lateral flow strip 5,6 uas muaj ob peb txheej ntawm cov porous nitrocellulose daim nyias nyias uas thawb ib qho me me ntawm cov qauv mus rau pem hauv ntej, reacting nrog pre-immobilized reagents los ntawm capillary quab yuam.Txawm hais tias lawv muaj qhov zoo ntawm tus nqi qis, kev siv yooj yim, thiab cov txiaj ntsig tau sai, cov khoom siv POCT siv tau tsuas yog siv rau kev sim tshuaj lom neeg (xws li, kuaj ntshav qabzib 7,8 thiab kuaj cev xeeb tub 9,10) yam tsis tas yuav tsum muaj kev tshuaj xyuas ntau theem.cov tshuaj tiv thaiv (xws li thauj khoom ntawm ntau cov tshuaj reagents, sib tov, multiplexing).Tsis tas li ntawd, cov tsav tsheb uas tswj cov dej txav (xws li, capillary rog) tsis muaj qhov sib xws zoo, tshwj xeeb tshaj yog nyob nruab nrab ntawm cov khoom siv, uas ua rau cov khoom siv tsis zoo11 thiab ua rau cov lateral flow bands feem ntau pab tau zoo rau kev kuaj pom 12,13.
Kev nthuav dav kev tsim khoom muaj peev xwm ntawm micro- thiab nanoscale tau tsim cov hauv kev rau kev txhim kho microfluidic POCT cov khoom siv rau kev ntsuas ntau 14,15,16,17.Los ntawm kev kho cov khoom ntawm lub interface 18, 19 thiab geometry ntawm cov channel 20, 21, 22, capillary quab yuam thiab ntws tus nqi ntawm cov khoom siv no tuaj yeem tswj tau.Txawm li cas los xij, lawv txoj kev ntseeg siab, tshwj xeeb tshaj yog rau cov kua dej uas ntub dej heev, tseem tsis tau txais vim yog kev tsim khoom tsis raug, cov khoom tsis xws luag, thiab rhiab heev rau ib puag ncig kev co.Tsis tas li ntawd, txij li thaum lub capillary khiav yog tsim los ntawm cov kua-gas interface, tsis muaj kev khiav dej num ntxiv, tshwj xeeb tshaj yog tom qab sau cov microfluidic channel nrog kua.Yog li ntawd, rau kev tshawb nrhiav ntau dua, ob peb kauj ruam ntawm kev txhaj tshuaj yuav tsum tau ua 24,25.
Ntawm cov khoom siv microfluidic, centrifugal microfluidic li tam sim no yog ib qho kev daws teeb meem zoo tshaj plaws rau POCT26,27.Nws txoj kev tsav tsheb yog qhov zoo nyob rau hauv uas lub zog tsav tsheb tuaj yeem tswj tau los ntawm kev kho cov kev sib hloov ceev.Txawm li cas los xij, qhov tsis zoo yog tias lub zog centrifugal yeej ib txwm coj mus rau sab nraud ntawm lub cuab yeej, ua rau nws nyuaj rau kev siv cov txheej txheem ntau yam uas yuav tsum tau ua rau kev tshuaj xyuas ntau dua.Txawm hais tias kev tsav tsheb ntxiv (xws li capillaries 28, 29 thiab ntau lwm tus 30, 31, 32, 33, 34, 35) ntxiv rau lub zog centrifugal tau qhia rau kev siv ntau yam, kev hloov pauv tsis tau pom dua tuaj yeem tshwm sim vim tias cov rog ntxiv no feem ntau raug txiav txim. ntawm qhov loj me qis dua lub zog centrifugal, ua rau lawv tsuas yog ua haujlwm zoo tshaj qhov kev ua haujlwm me me lossis tsis muaj nyob rau ntawm kev thov nrog cov kua tso tawm.Kev koom ua ke pneumatic manipulations rau hauv centrifugal microfluidics xws li centrifugal kinetic txoj kev 36, 37, 38, thermopneumatic txoj kev 39 thiab active pneumatic txoj kev 40 tau ua pov thawj tias yog ib qho kev ntxim nyiam lwm txoj kev.Nrog rau kev tawm tsam kev tawm tsam, ib qho kab noj hniav ntxiv thiab kev sib txuas microchannels tau muab tso rau hauv cov cuab yeej rau ob qho tib si sab nraud thiab sab hauv, txawm hais tias nws qhov kev ua tau zoo ntawm cov twj tso kua mis (hauv thaj tsam ntawm 75% mus rau 90%) yog nyob ntawm seb muaj pes tsawg lub voj voog thiab cov viscosity. ntawm cov kua.Nyob rau hauv txoj kev thermopneumatic, lub latex membrane thiab kua hloov chamber yog tsim tshwj xeeb los kaw los yog rov qhib lub inlet thaum lub ntim cua ntim tau rhuab los yog txias.Txawm li cas los xij, kev teeb tsa cua sov / cua txias qhia txog cov teeb meem teb qeeb thiab txwv tsis pub nws siv hauv cov kev ntsuas thermosensitive (piv txwv li, polymerase chain reaction (PCR) amplification).Nrog rau kev siv pneumatic mus kom ze, ntawm qhov kev thov tso tawm thiab sab hauv lub zog tau ua tiav los ntawm kev siv tib lub sijhawm ntawm lub siab zoo thiab sib tw sib tw nrawm los ntawm kev kub ceev motors.Muaj lwm txoj hauv kev zoo uas siv cov pneumatic actuators (zoo siab 41, 42 lossis siab tsis zoo 43) thiab feem ntau kaw valve tsim.Los ntawm kev ua tiav kev siv lub siab nyob rau hauv lub pneumatic chamber, cov kua yog pumped rau pem hauv ntej peristaltically, thiab ib txwm kaw valve tiv thaiv cov backflow ntawm cov kua vim peristalsis, yog li realizing complex kua ua hauj lwm.Txawm li cas los xij, tam sim no tsuas muaj tsawg tus lej ntawm cov thev naus laus zis microfluidic uas tuaj yeem ua haujlwm ua kua ua haujlwm nyuaj hauv ib qho khoom siv POCT, suav nrog ntau qhov kev faib tawm, kev tso tawm raws li qhov xav tau, kev ua haujlwm ruaj khov, kev cia ntev ntev, tuav cov kua dej siab viscosity, thiab raug nqi tsim khoom.Tag nrho tib lub sijhawm.Qhov tsis muaj kev ua haujlwm ntau kauj ruam kuj tseem yog ib qho laj thawj vim li cas tsuas yog qee qhov kev lag luam POCT cov khoom lag luam xws li Cepheid, Binx, Visby, Cobas Liat, thiab Rhonda tau ua tiav hauv kev lag luam qhib rau hnub tim.
Nyob rau hauv daim ntawv no, peb thov ib tug pneumatic microfluidic actuator raws li ntsuab nplhaib micro hloov tshuab (FAST).FAST muab tag nrho cov khoom tsim nyog nyob rau tib lub sijhawm rau ntau hom reagents los ntawm microliters rau milliliters.FAST muaj cov elastic membranes, levers thiab blocks.Yog tsis muaj daim ntawv thov ntawm huab cua siab, daim nyias nyias, levers thiab blocks tuaj yeem raug kaw nruj nreem thiab cov kua hauv tuaj yeem khaws cia rau lub sijhawm ntev.Thaum lub siab tsim nyog tau thov thiab hloov mus rau qhov ntev ntawm lub zog, lub diaphragm nthuav dav thiab thawb lub zog mus rau qhov qhib, tso cai rau cov kua dej dhau mus.Qhov no tso cai rau kev ntsuas dej ntau ntawm cov kua hauv ib qho cascade, ib txhij, ua ntu zus lossis xaiv yam.
Peb tau tsim ib lub PCR uas siv FAST los tsim cov lus teb-hauv-cov qauv ntsuas rau kev kuaj pom tus kab mob khaub thuas A thiab B (IAV thiab IBV).Peb tau ua tiav qhov kev txwv qis ntawm kev kuaj pom (LOD) ntawm 102 daim ntawv luam / mL, peb qhov kev ntsuam xyuas multiplex tau pom qhov tshwj xeeb rau IAV thiab IBV thiab tso cai rau tus kab mob khaub thuas kab mob.Cov txiaj ntsig kev soj ntsuam tau siv lub qhov ntswg swab qauv los ntawm 18 tus neeg mob thiab 18 tus neeg noj qab haus huv qhia pom tias muaj kev sib raug zoo hauv fluorescence siv nrog tus qauv RT-PCR (Pearson coefficients> 0.9).Cov txiaj ntsig kev soj ntsuam tau siv lub qhov ntswg swab qauv los ntawm 18 tus neeg mob thiab 18 tus neeg noj qab haus huv qhia pom tias muaj kev sib raug zoo hauv fluorescence siv nrog tus qauv RT-PCR (Pearson coefficients> 0.9).Результаты клинических испытаний с использованием образца мазка из носа от 18 пациентов и 18 здоровых тствие интенсивности флуоресценции стандартной ОТ-ПЦР (коэффициенты Пирсона > 0,9).Cov txiaj ntsig ntawm kev sim tshuaj siv lub qhov ntswg qhov ntswg los ntawm 18 tus neeg mob thiab 18 tus neeg noj qab haus huv pom zoo pom zoo ntawm fluorescence siv ntawm tus qauv RT-PCR (Pearson's coefficients> 0.9).0.9) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Результаты клинических испытаний с использованием образцов назальных мазков от 18 пациентов и 18 зкадлировых ствие между интенсивностью флуоресценции и стандартной ОТ-ПЦР (коэффициент Пирсона > 0,9).Cov txiaj ntsig ntawm kev sim tshuaj siv qhov ntswg qhov ntswg los ntawm 18 tus neeg mob thiab 18 tus neeg noj qab haus huv tau pom zoo pom zoo ntawm fluorescence siv thiab tus qauv RT-PCR (Pearson's coefficient> 0.9).Tus nqi kwv yees ntawm cov khoom siv FAST-POCT yog kwv yees li US $ 1 (Table Ntxiv 1) thiab tuaj yeem txo qis ntxiv los ntawm kev siv cov txheej txheem tsim khoom loj (xws li txhaj tshuaj molding).Qhov tseeb, FAST-based POCT cov cuab yeej muaj tag nrho cov yam ntxwv tsim nyog tau txib los ntawm WHO thiab tau sib xws nrog cov kev sim tshuaj biochemical tshiab xws li plasma thermal cycling44, amplification-free immunoassays45 and nanobody functionalization tests46 that are the backbone of POCT systems.muaj peev xwm.
Ntawm daim duab.1a qhia cov qauv ntawm FAST-POCT platform, uas muaj plaub chav ua kua: chav ua ntej cia, chav sib xyaw, chav tshuaj tiv thaiv, thiab cov khib nyiab chamber.Tus yuam sij rau kev tswj cov dej ntws yog qhov tsim qauv FAST (suav nrog elastic membranes, levers thiab blocks) nyob rau hauv pre-storage chamber thiab mixing chamber.Raws li cov txheej txheem pneumatically actuated, FAST tsim muab kev tswj cov kua dej meej, suav nrog kev kaw / qhib kev hloov pauv, kev siv ntau yam, kev tso tawm cov kua dej raws li qhov xav tau, kev ua haujlwm txhim khu kev qha (xws li, tsis hnov tsw rau ib puag ncig kev co), thiab khaws cia ntev.Lub FAST-POCT platform muaj plaub txheej: txheej txheej rov qab, txheej txheej zaj duab xis elastic, txheej zaj duab xis yas, thiab txheej txheej npog, raws li pom nyob rau hauv daim duab loj hauv daim duab 1b (tseem qhia kom meej hauv cov duab ntxiv S1 thiab S2. ).Tag nrho cov channel thiab cov khoom thauj cov kua dej (xws li cov khoom ntim ua ntej thiab cov tshuaj tiv thaiv kab mob) yog kos rau hauv PLA (polylactic acid) substrates xws li 0.2 hli (thinnest feem) mus rau 5 hli tuab.Cov khoom siv zaj duab xis elastic yog 300 µm tuab PDMS uas yooj yim nthuav thaum huab cua siab siv vim nws "nyias tuab" thiab qis modulus ntawm elasticity (txog 2.25 MPa47).Polyethylene zaj duab xis txheej yog ua los ntawm polyethylene terephthalate (PET) nrog ib tug tuab ntawm 100 µm los tiv thaiv cov elastic zaj duab xis los ntawm ntau deformation vim huab cua siab.Ua ke rau cov chav, cov txheej txheej substrate muaj cov levers txuas nrog lub npog txheej (ua los ntawm PLA) los ntawm cov pob khawm los tswj cov dej ntws.Cov zaj duab xis elastic tau muab khi rau hauv cov txheej txheem rov qab uas siv ob daim kab xev nplaum (ARseal 90880) thiab npog nrog ib zaj duab xis yas.Peb txheej tau sib sau ua ke ntawm lub substrate siv T-clip tsim nyob rau hauv lub npog txheej.Lub T-clamp muaj qhov sib txawv ntawm ob txhais ceg.Thaum cov clip tau muab tso rau hauv qhov zawj, ob txhais ceg khoov me ntsis, tom qab ntawd rov qab mus rau lawv qhov qub thiab nruj nreem khi lub hau thiab nraub qaum thaum lawv dhau los ntawm qhov zawj (Sab ntxiv Fig. S1).Plaub txheej yog ces sib sau ua ke siv connectors.
Schematic daim duab ntawm lub platform qhia txog ntau yam kev ua haujlwm thiab cov yam ntxwv ntawm FAST.b Daim duab loj ntawm FAST-POCT platform.c Diam duab ntawm lub platform nyob ib sab ntawm ib lub quarter duas nyiaj npib.
Cov txheej txheem ua haujlwm ntawm FAST-POCT platform yog qhia hauv daim duab 2. Cov khoom tseem ceeb yog cov blocks ntawm lub hauv paus txheej thiab cov hinges ntawm lub npog txheej, uas ua rau muaj kev cuam tshuam tsim thaum plaub txheej yog sib sau ua ke siv T-zoo li. .Thaum tsis muaj huab cua siab siv (daim duab 2a), qhov cuam tshuam haum ua rau lub pob khawm khoov thiab deform, thiab lub sealing quab yuam yog siv los ntawm lub lever los nias lub elastic zaj duab xis tiv thaiv lub thaiv, thiab cov kua hauv lub seal kab noj hniav yog txhais. raws li lub xeev kaw.Nws yuav tsum tau muab sau tseg tias nyob rau hauv lub xeev no, lub lever yog khoov rau sab nraud, raws li qhia nyob rau hauv lub sab saib nyob rau hauv daim duab 2a.Thaum huab cua nkag (Fig. 2b), lub elastic membrane nthuav tawm sab nraud ntawm lub hau thiab thawb lub zog, yog li qhib qhov sib txawv ntawm lub zog thiab qhov thaiv kom cov dej ntws mus rau hauv lub chamber tom ntej, uas txhais tau tias yog lub xeev qhib. .Tom qab tso tawm ntawm huab cua siab, lub zog tuaj yeem rov qab mus rau nws txoj haujlwm qub thiab nyob twj ywm nruj vim qhov elasticity ntawm pob khawm.Cov yeeb yaj kiab ntawm lub zog txav tau nthuav tawm hauv cov yeeb yaj kiab ntxiv S1.
A. Schematic daim duab thiab cov duab thaum kaw.Thaum tsis muaj lub siab, lub lever presses lub membrane tiv thaiv lub thaiv, thiab cov kua yog kaw.b Nyob zoo.Thaum lub siab yog thov, daim nyias nyias nthuav thiab thawb lub zog, yog li cov channel qhib thiab cov kua dej tuaj yeem ntws.c Txiav txim siab tus yam ntxwv loj ntawm lub siab tseem ceeb.Cov yam ntxwv qhov ntev muaj xws li qhov ntev ntawm lub lever (L), qhov kev ncua deb ntawm lub slider thiab lub pob khawm (l) thiab lub thickness ntawm lub lever protrusion (t).Fs yog lub compaction quab yuam ntawm lub throttle point B. q yog lub uniformly faib load ntawm lub lever.Tx* sawv cev rau lub torque tsim los ntawm hinged lever.Lub siab tseem ceeb yog lub siab xav tau los tsa lub zog thiab ua kom cov kua dej ntws tawm.d Theoretical thiab kev sim cov txiaj ntsig ntawm kev sib raug zoo ntawm qhov tseem ceeb ntawm lub siab thiab lub ntsiab lus loj.n = 6 kev sim ywj pheej tau ua thiab cov ntaub ntawv tau pom tias yog ± tus qauv sib txawv.Cov ntaub ntawv nyoos yog nthuav tawm raws li cov ntaub ntawv raw cov ntaub ntawv.
Ib qho qauv tshuaj ntsuam xyuas raws li txoj kev xav ntawm kab teeb tau tsim los txheeb xyuas qhov kev cia siab ntawm qhov tseem ceeb Pc ntawm qhov sib txawv ntawm qhov tsis sib xws (piv txwv li, L yog qhov ntev ntawm lub zog, l yog qhov kev ncua deb ntawm qhov thaiv thiab qhov sib txawv. pob khawm, S yog lub lever Qhov chaw sib cuag nrog cov kua t yog lub thickness ntawm lub lever protrusion, raws li qhia nyob rau hauv daim duab 2c).Raws li tau piav qhia hauv Cov Lus Qhia Ntxiv thiab Cov Duab Ntxiv S3, qhov sib txawv qhib thaum \({P}_{c}\ge \frac{2{F}_{s}l}{SL}\), qhov twg Fs yog lub zog. \ ({T}_{x}^{\ast}(={F}_{s}l)\) tshem tawm cov rog uas cuam tshuam nrog kev cuam tshuam thiab ua rau lub pob khawm khoov.Cov lus teb rau kev sim thiab cov qauv kev soj ntsuam qhia pom pom zoo (Daim duab 2d), qhia tias qhov tseem ceeb ntawm Pc nce nrog nce t / l thiab txo qis L, uas tau yooj yim piav qhia los ntawm cov qauv beam classic, piv txwv li lub torque nce nrog t / Lift. .Yog li, peb qhov kev tshawb xav tau qhia meej meej tias qhov tseem ceeb ntawm lub siab tuaj yeem tswj tau zoo los ntawm kev kho lub zog ntev L thiab t / l piv, uas muab lub hauv paus tseem ceeb rau kev tsim ntawm FAST-POCT platform.
Lub FAST-POCT platform muab ntau qhov kev faib tawm (pom hauv daim duab 3a nrog inset thiab sim), uas yog qhov tseem ceeb tshaj plaws ntawm kev ua tiav POCT, qhov twg cov kua dej tuaj yeem ntws mus rau txhua qhov kev taw qhia thiab nyob rau hauv ib qho kev txiav txim (cascade, ib txhij, sequential) los yog xaiv multichannel. muab tshuaj.- dosing muaj nuj nqi.Ntawm daim duab.3a(i) qhia tau hais tias cascaded dosing hom nyob rau hauv uas ob los yog ntau lub chamber yog cascaded siv blocks cais cov ntau yam reactants thiab ib tug lever los tswj lub qhib thiab kaw lub xeev.Thaum lub siab yog thov, cov kua ntws los ntawm lub sab sauv mus rau lub qis chamber nyob rau hauv ib tug cascade yam.Nws yuav tsum tau muab sau tseg tias lub cascade chambers tuaj yeem sau nrog cov tshuaj ntub dej lossis cov tshuaj qhuav xws li cov hmoov lyophilized.Nyob rau hauv qhov kev sim hauv daim duab 3a(i), tus number case liab los ntawm lub chamber sab sauv ntws nrog rau cov xim xiav dye hmoov (tooj liab sulfate) mus rau hauv lub chamber thib ob thiab tig xiav tsaus thaum nws mus txog lub chamber qis.Nws kuj qhia tau hais tias kev tswj siab rau cov kua dej uas tau muab tso rau.Ib yam li ntawd, thaum ib lub zog txuas nrog ob chav, nws dhau los ua hom kev txhaj tshuaj ib txhij, raws li qhia hauv daim duab.3a(ii), nyob rau hauv uas cov kua yuav tsum uniformly faib nyob rau hauv ob los yog ntau lub chamber thaum lub siab siv.Txij li thaum lub siab tseem ceeb nyob ntawm qhov ntev ntawm lub zog, qhov ntev ntawm lub zog tuaj yeem hloov kho kom ua tiav cov qauv txhaj tshuaj raws li qhia hauv daim duab.3 a(iii).Lub pas nrig ntev (nrog rau lub siab tseem ceeb Pc_long) tau txuas nrog lub chamber B thiab lub kauj luv luv (nrog rau lub siab tseem ceeb Pc_short> Pc_long) tau txuas nrog lub chamber A. Raws li lub siab P1 (Pc_long < P1 < Pc_short) tau siv, tsuas yog cov kua dej liab. tuaj yeem ntws mus rau chamber B thiab thaum lub siab tau nce mus rau P2 (> Pc_short), cov kua xiav tuaj yeem ntws mus rau chamber A. Hom kev txhaj tshuaj no siv tau rau cov kua sib txawv hloov mus rau lawv cov chav sib txuas ua ke, uas yog qhov tseem ceeb rau kev ua tiav POCT. ntaus ntawv.Lub pas nrig ntev (nrog rau lub siab tseem ceeb Pc_long) tau txuas nrog lub chamber B thiab lub kauj luv luv (nrog rau lub siab tseem ceeb Pc_short> Pc_long) tau txuas nrog lub chamber A. Raws li lub siab P1 (Pc_long < P1 < Pc_short) tau siv, tsuas yog cov kua dej liab. tuaj yeem ntws mus rau chamber B thiab thaum lub siab tau nce mus rau P2 (> Pc_short), cov kua xiav tuaj yeem ntws mus rau chamber A. Hom kev txhaj tshuaj no siv tau rau cov kua sib txawv hloov mus rau lawv cov chav sib txuas ua ke, uas yog qhov tseem ceeb rau kev ua tiav POCT. ntaus ntawv.Длинный рычаг (с критическим давлением Pc_long) был соединен с камерой B, а короткий рычаг (с критическим) соединен с камерой A. При приложении давления P1 (Pc_long < P1 < Pc_short) только жидкость, выделенная красьны м камеру B, и когда давление было увеличено до P2 (> Pc_short), синяя жидкость может течь в камеров A. Эльотрожет применяется к различным жидкостям, последовательно перемещаемым в соответствующие дмеры, что ичешет tsis yog POCT.Lub pas nrig ntev (nrog rau lub siab tseem ceeb Pc_long) tau txuas nrog lub chamber B, thiab lub kauj luv luv (nrog rau lub siab tseem ceeb Pc_short> Pc_long) tau txuas nrog rau chav A. Thaum lub siab P1 (Pc_long < P1 < Pc_short) yog siv, tsuas yog cov kua uas tseem ceeb. nyob rau hauv liab tuaj yeem ntws mus rau hauv chamber B, thiab thaum lub siab tau nce mus rau P2 (> Pc_short), cov kua xiav tuaj yeem ntws mus rau hauv chamber A. Hom kev txhaj tshuaj no yog siv rau cov kua sib txawv sib txuas mus rau lwm chav, uas yog qhov tseem ceeb. rau kev ua tiav POCT.ntaus ntawv. Длинный рычаг (критическое давление Pc_long) соединен с камерой B, а короткий рычаг (критическое давлени давлени > Pc рой A.Lub caj npab ntev (lub siab tseem ceeb Pc_long) txuas nrog lub chamber B thiab lub caj npab luv (lub siab tseem ceeb Pc_short> Pc_long) txuas nrog rau chav A.При приложении давления P1 (Pc_long < P1 < Pc_short) в камеру B может поступать только красная жидкость , Pв приние _short) в камеру A может поступать синяя жидкость.Thaum lub siab P1 (Pc_long < P1 < Pc_short) yog siv, tsuas yog cov kua liab tuaj yeem nkag mus rau hauv chav B, thiab thaum lub siab nce mus rau P2 (> Pc_short), cov kua xiav tuaj yeem nkag mus rau hauv chav A. Qhov kev txhaj tshuaj ua ntu zus no yog tsim rau kev hloov pauv hloov mus ua ntu zus. ntau yam kua rau hauv cov chav, uas yog ib qho tseem ceeb rau kev ua tau zoo ntawm cov khoom siv POCT.Daim duab 3a(iv) qhia txog cov kev xaiv txhaj tshuaj, qhov twg lub chamber tseem ceeb muaj luv luv (nrog rau lub siab tseem ceeb Pc_short) thiab lub zog ntev (nrog rau lub siab tseem ceeb Pc_long < Pc_short) uas tau txuas nrog chamber A thiab chamber B, feem, ntxiv rau. mus rau lwm qhov cua channel txuas nrog chamber B. Txhawm rau hloov cov kua rau chamber A thawj zaug, siab P1 (Pc_long < P1 < Pc_short) thiab P2 (P2 > P1) nrog P1 + P2 > Pc_short tau siv rau lub cuab yeej tib lub sijhawm.Daim duab 3a(iv) qhia txog cov kev xaiv txhaj tshuaj, qhov twg lub chamber tseem ceeb muaj luv luv (nrog rau lub siab tseem ceeb Pc_short) thiab lub zog ntev (nrog rau lub siab tseem ceeb Pc_long < Pc_short) uas tau txuas nrog chamber A thiab chamber B, feem, ntxiv rau. mus rau lwm qhov cua channel txuas nrog chamber B. Txhawm rau hloov cov kua rau chamber A thawj zaug, siab P1 (Pc_long < P1 < Pc_short) thiab P2 (P2 > P1) nrog P1 + P2 > Pc_short tau siv rau lub cuab yeej tib lub sijhawm.Ntawm daim duab.3а(iv) показан режим селективного впрыска, при котором основная камера имела короткий (с критический даc критический даc аг (с критическим давлением Pc_long < Pc_short), которые дополнительно соединялись с камерой A и камерой B свет.3a(iv) qhia txog cov kev xaiv txhaj tshuaj, uas lub ntsiab chamber muaj luv luv (nrog rau lub siab tseem ceeb Pc_short) thiab ib tug ntev lever (nrog rau qhov tseem ceeb siab Pc_long < Pc_short), uas tau txuas ntxiv rau chamber A thiab chamber B, feem.к другому воздушному каналу, соединенному с камерой B. Чтобы сначала передать жидкость в камерустр онумеруса, икладывали давление P1 (Pc_long < P1 < Pc_short) и P2 (P2 > P1), где P1 + P2 > Pc_short.mus rau lwm cov huab cua channel txuas nrog chamber B. Txhawm rau ua ntej hloov cov kua rau chamber A, pressures P1 (Pc_long < P1 < Pc_short) thiab P2 (P2 > P1) tau siv ib txhij rau lub cuab yeej, qhov twg P1 + P2 > Pc_short. 3а(iv) показан режим селективного впрыска, когда основная камера имеет короткий стержень (с критическим с критическим стержень (с критическим давлением Pc_long < Pc_short), соединенные с камерой A и камерой B соответствоннно, и в урнимерой B соответствоншно kuv mas, подключенному к комнате B.3a(iv) qhia txog kev xaiv txhaj tshuaj thaum lub ntsiab chamber muaj lub qia luv (qhov tseem ceeb siab Pc_short) thiab lub qia ntev (lub siab tseem ceeb Pc_long < Pc_short) txuas nrog rau chav A thiab chav B feem, thiab ntxiv rau lwm qhov cua, txuas rau chav B.Yog li, P2 tiv thaiv cov kua nkag mus rau hauv chav B;Lub sijhawm no, tag nrho cov siab P1 + P2 tau dhau qhov tseem ceeb tshaj plaws kom qhib lub qis qis txuas nrog chamber A kom tso cai rau cov kua ntws mus rau chav A. Tom qab ntawd, thaum chamber B yuav tsum tau ntim, peb tsuas yog yuav tsum tau thov P1 (Pc_long < P1 < Pc_short) nyob rau hauv lub ntsiab chamber kom qhib lub siab ntev thiab tso cai rau cov kua ntws mus rau chamber B. Nws tuaj yeem pom meej txij lub sijhawm t = 3 s txog 9 s tias cov kua hauv chamber A tseem nyob tas li thaum nws nce hauv chamber. B thaum siv lub siab P1.Lub sijhawm no, tag nrho cov siab P1 + P2 tau dhau qhov tseem ceeb tshaj plaws kom qhib lub qis qis txuas nrog chamber A kom tso cai rau cov kua ntws mus rau chav A. Tom qab ntawd, thaum chamber B yuav tsum tau ntim, peb tsuas yog yuav tsum tau thov P1 (Pc_long < P1 < Pc_short) nyob rau hauv lub ntsiab chamber kom qhib lub siab ntev thiab tso cai rau cov kua ntws mus rau chamber B. Nws tuaj yeem pom meej txij lub sijhawm t = 3 s txog 9 s tias cov kua hauv chamber A tseem nyob tas li thaum nws nce hauv chamber. B thaum siv lub siab P1.Между тем, общее давление P1 + P2 превысило критическое давление, чтобы активировать более короткагий сы чтобы позволить жидкости течь в камеру A. Затем, когда требуется заполнить камеру B, нам нужно толькько полько полько ) в основной камере, чтобы активировать длинный рычаг и дать жидкости течь в камеру B. Можно яснть на дла да с до 9 с жидкость в камере A оставалась постоянной, в то время как в камере она увеличивалась.Lub caij no, tag nrho lub siab P1 + P2 tau tshaj qhov tseem ceeb ntawm lub siab kom qhib lub kauj luv luv txuas nrog chamber A kom cov kua ntws mus rau hauv chamber A. Tom qab ntawd thaum chamber B yuav tsum tau ntim, peb tsuas yog yuav tsum tau thov P1 (Pc_long
Ib qho piv txwv ntawm ntau txoj haujlwm, xws li (i) cascading, (ii) ib txhij, (iii) ua ntu zus, thiab (iv) xaiv txoj haujlwm.Cov curves sawv cev rau kev ua haujlwm thiab cov kev txwv ntawm plaub hom kev faib tawm no.b Cov txiaj ntsig ntawm kev sim cia ntev ntev hauv dej deionized thiab ethanol.n = 5 qhov kev sim ywj pheej tau ua thiab cov ntaub ntawv qhia tias ± sd c.Kev ntsuas kev ruaj ntseg ua qauv qhia thaum lub FAST ntaus ntawv thiab lub capillary valve (CV) ntaus ntawv nyob rau hauv (i) zoo li qub thiab (ii) vibrating xeev.(iii) Volume vs. time for Fast and CV devices at various angular frequencies.d Kev tshaj tawm cov txiaj ntsig kev xeem ntawm kev thov rau (i) FAST ntaus ntawv thiab (ii) CV ntaus ntawv.(iii) Kev sib raug zoo ntawm ntim thiab lub sijhawm rau FAST thiab CV li siv hom kev sib tshuam.Tag nrho cov scale bar, 1 cm.Cov ntaub ntawv nyoos yog muab raws li cov ntaub ntawv raw cov ntaub ntawv.
Kev khaws cia ntev ntev ntawm cov tshuaj reagents yog lwm qhov tseem ceeb ntawm kev ua tiav POCT cov cuab yeej uas yuav tso cai rau cov neeg ua haujlwm tsis tau txais kev pabcuam los tuav ntau cov tshuaj reagents.Txawm hais tias muaj ntau yam thev naus laus zis tau qhia lawv lub peev xwm rau kev khaws cia ntev (xws li, 35 microdispensers, 48 blister packs, thiab 49 stick packs), yuav tsum muaj cov khoom ntim khoom tshwj xeeb kom haum rau cov pob khoom, uas ua rau cov nqi thiab qhov nyuaj;Tsis tas li ntawd, cov txheej txheem khaws cia no tsis tso cai rau kev xa tawm raws li qhov xav tau thiab ua rau cov khoom pov tseg vim cov khoom seem hauv lub ntim.Lub peev xwm khaws cia ntev tau raug txheeb xyuas los ntawm kev ntsuas lub neej nrawm siv cov khoom siv CNC-machined PMMA vim nws qhov roughness me ntsis thiab tsis kam rau cov pa permeation (Cov duab ntxiv S5).Cov cuab yeej ntsuas tau ntim nrog dej deionized (deionized dej) thiab 70% ethanol (simulating volatile reagents) ntawm 65 ° C rau 9 hnub.Ob qho dej deionized thiab ethanol tau khaws cia siv cov ntawv txhuas los thaiv kev nkag los ntawm saum toj no.Qhov sib npaug Arrhenius thiab nkag mus ua kom lub zog tau tshaj tawm hauv cov ntaub ntawv50,51 tau siv los xam qhov sib npaug ntawm lub sijhawm.Ntawm daim duab.3b qhia qhov nruab nrab qhov hnyav poob rau 5 cov qauv khaws cia ntawm 65 ° C rau 9 hnub, sib npaug rau 0.30% rau cov dej deionized thiab 0.72% rau 70% ethanol tshaj 2 xyoo ntawm 23 ° C.
Ntawm daim duab.3c qhia kev vibration test.Txij li thaum lub capillary valve (CV) yog txoj kev siv dej nrov tshaj plaws ntawm cov khoom siv POCT28,29 uas twb muaj lawm, CV ntaus ntawv 300 µm dav thiab 200 µm sib sib zog nqus tau siv rau kev sib piv.Nws tuaj yeem pom tau tias thaum ob qho khoom siv nyob ruaj ruaj, cov kua dej hauv FAST-POCT platform ntsaws ruaj ruaj thiab cov kua dej hauv CV ntaus ntawv kaw vim qhov kev nthuav dav ntawm cov channel, uas txo cov capillary rog.Txawm li cas los xij, raws li lub orbital vibrator lub angular zaus nce, cov kua hauv lub FAST-POCT platform tseem kaw, tab sis cov kua dej hauv CV ntaus ntawv ntws mus rau hauv lub chamber qis (saib Ntxiv Movie S3).Qhov no qhia tau hais tias lub pob khawm deformable ntawm FAST-POCT platform tuaj yeem siv lub zog txhua yam rau lub module kom kaw cov kua hauv chamber.Txawm li cas los xij, hauv CV cov khoom siv, cov kua tau khaws cia vim qhov sib npaug ntawm cov khoom, huab cua, thiab cov theem ua kua, tsim kev tsis ruaj khov, thiab kev vibrations tuaj yeem cuam tshuam qhov sib npaug thiab ua rau tus cwj pwm tsis txaus ntseeg.Qhov kom zoo dua ntawm FAST-POCT platform yog tias nws muab kev ua haujlwm txhim khu kev qha thiab zam kev ua tsis tiav thaum muaj kev vibrations uas feem ntau tshwm sim thaum xa khoom thiab ua haujlwm.
Lwm qhov tseem ceeb ntawm FAST-POCT platform yog nws qhov kev thov tso tawm, uas yog qhov tseem ceeb rau kev txheeb xyuas ntau yam.Ntawm daim duab.3d sib piv qhov kev thov tso tawm ntawm FAST-POCT platform thiab CV ntaus ntawv.Los ntawm fig.3d(iii) peb pom tias cov cuab yeej FAST teb sai sai rau cov teeb liab siab.Thaum lub siab tau thov rau FAST-POCT platform, cov kua dej ntws tawm, thaum lub siab tso tawm, qhov ntws tam sim ntawd nres (Daim duab 3d(i)).Qhov kev txiav txim no tuaj yeem piav qhia los ntawm qhov nrawm nrawm rov qab ntawm lub pob khawm, uas nias lub zog rov qab tiv thaiv qhov thaiv, kaw lub chamber.Txawm li cas los xij, cov kua dej txuas ntxiv mus rau hauv cov cuab yeej CV, thaum kawg ua rau cov kua dej tsis txaus ntseeg ntawm kwv yees li 100 µl tom qab lub siab tso tawm (Daim duab 3d (ii) thiab Ntxiv Movie S4).Qhov no tuaj yeem piav qhia los ntawm kev ploj mus ntawm capillary pinning nyhuv thaum ua tiav ntub dej ntawm CV tom qab thawj zaug txhaj tshuaj.
Lub peev xwm los tswj cov kua dej sib txawv ntawm cov dej sib txawv thiab cov viscosity hauv tib lub cuab yeej tseem yog qhov nyuaj rau POCT daim ntawv thov.Kev ntub dej tsis zoo tuaj yeem ua rau cov xau lossis lwm yam kev xav tsis txaus ntseeg hauv cov channel, thiab cov khoom siv ntxiv xws li vortex mixers, centrifuges thiab cov ntxaij lim dej feem ntau yuav tsum tau npaj cov kua dej viscous 52.Peb tau soj ntsuam cov kev sib raug zoo ntawm lub siab tseem ceeb thiab cov khoom ua kua (nrog rau ntau yam ntawm wettability thiab viscosity).Cov txiaj ntsig tau pom hauv Table 1 thiab Video S5.Nws tuaj yeem pom tias cov kua dej sib txawv ntawm qhov sib txawv ntawm qhov sib txawv thiab cov viscosity tuaj yeem muab ntim rau hauv lub chamber, thiab thaum siv lub siab, txawm tias cov kua dej uas muaj viscosity txog li 5500 cP tuaj yeem hloov mus rau lub chamber uas nyob ib sab, ua rau nws tuaj yeem ntes cov qauv nrog siab. viscosity (piv txwv li hnoos qeev, ib qho piv txwv viscous siv rau kev kuaj mob ntawm cov kab mob ua pa).
Los ntawm kev sib txuas cov khoom siv ntau yam khoom siv saum toj no, ntau yam ntawm FAST-based POCT cov cuab yeej tuaj yeem tsim tau.Ib qho piv txwv yog muaj nyob rau hauv daim duab 1. Cov nroj tsuag muaj lub pre-storage chamber, ib tug tov chamber, cov tshuaj tiv thaiv chamber thiab ib tug khib chamber.Reagents tuaj yeem khaws cia rau hauv chav ua ntej khaws cia rau lub sijhawm ntev thiab tom qab ntawd tawm mus rau hauv chav sib xyaw.Nrog rau txoj cai siab, cov tshuaj sib xyaw ua ke tuaj yeem xaiv tau hloov mus rau hauv cov khib nyiab los yog cov tshuaj tiv thaiv chamber.
Vim tias PCR kuaj pom yog tus qauv kub rau kev kuaj xyuas cov kab mob xws li H1N1 thiab COVID-19 thiab koom nrog ntau cov kauj ruam tshuaj tiv thaiv, peb siv FAST-POCT platform rau PCR tshawb pom raws li daim ntawv thov.Ntawm daim duab.4 qhia txog cov txheej txheem kuaj PCR siv FAST-POCT platform.Ua ntej, cov eluting reagent, sib nqus microbead reagent, ntxuav kua A, thiab ntxuav kua W tau pipetted rau hauv pre-storage chambers E, M, W1 thiab W2, raws li.Cov theem ntawm RNA adsorption yog qhia hauv daim duab.4a thiab muaj raws li hauv qab no: (1) thaum siab P1 (= 0.26 bar) yog siv, cov qauv txav mus rau hauv chamber M thiab tawm mus rau hauv chav sib tov.(2) Huab cua siab P2 (= 0.12 bar) yog muab los ntawm qhov chaw nres nkoj A txuas rau hauv qab ntawm chav sib xyaw.Txawm hais tias muaj ntau txoj kev sib xyaw ua ke tau qhia lawv lub peev xwm hauv kev sib xyaw ua kua ntawm POCT platforms (xws li serpentine mixing 53, random mixing 54 thiab batch mixing 55), lawv cov kev sib xyaw ua haujlwm thiab kev ua tau zoo tseem tsis txaus siab.Nws siv txoj kev sib xyaw ua npuas dej, uas cov cua nkag mus rau hauv qab ntawm chav sib xyaw kom tsim cov npuas hauv cov kua, tom qab ntawd cov vortex muaj zog tuaj yeem ua tiav kev sib xyaw hauv vib nas this.Kev sim ua npuas sib xyaw tau ua tiav thiab cov txiaj ntsig tau nthuav tawm hauv Daim Duab Ntxiv S6.Nws tuaj yeem pom tau tias thaum lub siab ntawm 0.10 bar tau thov, kev sib xyaw ua tiav yuav siv li 8 vib nas this.Los ntawm kev nce siab rau 0.20 bar, ua tiav kev sib xyaw ua tiav hauv li 2 vib nas this.Cov txheej txheem rau kev suav kev sib xyaw ua haujlwm tau nthuav tawm hauv Cov Txheej Txheem.(3) Siv cov hlau nplaum rubidium los rho cov hlaws dai, ces nias P3 (= 0.17 bar) los ntawm qhov chaw nres nkoj P kom txav cov reagents mus rau hauv cov khib nyiab chamber.Ntawm daim duab.4b, c qhia cov kauj ruam ntxuav kom tshem tawm cov impurities los ntawm cov qauv hauv qab no: (1) Cov tshuaj ntxuav dej A los ntawm lub chamber W1 yog tawm mus rau hauv lub siab sib xyaw chamber P1.(2) Tom qab ntawd ua cov txheej txheem npuas sib xyaw.(3) Lub tshuab ntxhua khaub ncaws A raug xa mus rau cov khib nyiab kua chamber, thiab cov microbeads hauv chav sib xyaw yog rub tawm los ntawm cov hlau nplaum.Ntxuav W (Fig. 4c) zoo ib yam li ntxuav A (Fig. 4b).Nws yuav tsum raug sau tseg tias txhua kauj ruam A thiab W tau ua tiav ob zaug.Daim duab 4d qhia cov kauj ruam elution los elute RNA los ntawm cov hlaws dai;cov elution thiab mixing taw qhia cov kauj ruam yog tib yam li RNA adsorption thiab ntxuav cov kauj ruam tau piav saum toj no.Raws li cov elution reagents raug xa mus rau hauv PCR cov tshuaj tiv thaiv chamber nyob rau hauv lub siab P3 thiab P4 (= 0.23 bar), lub siab tseem ceeb tau mus txog rau khi caj npab ntawm PCR cov tshuaj tiv thaiv chamber.Ib yam li ntawd, lub siab P4 kuj tseem pab txhawm rau txhawm rau txhawm rau nkag mus rau hauv cov khib nyiab chamber.Yog li, tag nrho cov elution reagents tau muab faib sib npaug ntawm plaub lub PCR cov tshuaj tiv thaiv kab mob los pib qhov sib txawv PCR cov tshuaj tiv thaiv.Cov txheej txheem saum toj no tau nthuav tawm hauv Kev Ua Yeeb Yam Ntxiv S6.
Nyob rau hauv RNA adsorption kauj ruam, tus qauv yog nkag mus rau hauv lub qhov nkag M thiab txhaj rau hauv chav sib xyaw nrog rau cov hlaws khaws cia yav dhau los.Tom qab sib tov thiab tshem tawm cov granules, cov reagents muab faib rau hauv cov khib nyiab chamber.b thiab c ntxuav cov kauj ruam, qhia ntau yam ua ntej khaws cov tshuaj reagents rau hauv chav sib tov, thiab tom qab sib tov thiab tshem cov hlaws dai, hloov cov reagents mus rau cov khib nyiab kua chamber.d Elution kauj ruam: Tom qab qhia cov elution reagents, sib tov thiab hlaws rho tawm, cov reagents raug xa mus rau PCR cov tshuaj tiv thaiv chamber.Cov nkhaus qhia txog kev ua haujlwm thiab kev cuam tshuam ntawm ntau theem.Lub siab yog lub siab exerted los ntawm tus kheej chav.Volume yog lub ntim ntawm cov kua nyob rau hauv lub mixing chamber.Tag nrho cov scale bar yog 1 cm.Cov ntaub ntawv nyoos yog muab raws li cov ntaub ntawv raw cov ntaub ntawv.
Cov txheej txheem kuaj PCR tau ua tiav thiab Cov Duab Ntxiv S7 nthuav qhia cov ntaub ntawv thermal nrog rau 20 feeb ntawm lub sijhawm rov qab thiab 60 feeb ntawm lub sijhawm thermal cycling (95 thiab 60 ° C), nrog rau ib lub voj voog thermal yog 90 s (Suplementary Movie S7)..FAST-POCT yuav tsum muaj sijhawm tsawg dua los ua kom tiav ib lub voj voog thermal (90 vib nas this) dua li cov qauv RT-PCR (180 vib nas this rau ib lub voj voog thermal).Qhov no tuaj yeem piav qhia los ntawm thaj chaw siab rau qhov ntim piv thiab qhov qis thermal inertia ntawm micro-PCR cov tshuaj tiv thaiv chamber.Lub chamber nto yog 96.6 mm2 thiab lub chamber ntim yog 25 mm3, ua rau saum npoo rau ntim piv kwv yees li 3.86.Raws li pom hauv Daim Duab Ntxiv S10, PCR qhov chaw kuaj ntawm peb lub platform muaj qhov zawj ntawm lub vaj huam sib luag tom qab, ua rau hauv qab ntawm PCR chamber 200 µm tuab.Ib lub thermally conductive elastic ncoo yog txuas rau lub tshav kub ntawm lub ntsuas kub ntawm lub ntsuas kub, xyuas kom nruj kev sib cuag nrog rau sab nraum qab ntawm lub thawv kuaj.Qhov no txo cov thermal inertia ntawm lub platform thiab txhim kho cov cua sov / txias efficiency.Thaum lub sij hawm thermal cycling, cov paraffin embedded nyob rau hauv lub platform melts thiab ntws mus rau hauv lub PCR cov tshuaj tiv thaiv chamber, ua raws li ib tug sealant los tiv thaiv reagent evaporation thiab ib puag ncig paug (saib Supplementary Movie S8).
Tag nrho cov txheej txheem kuaj xyuas PCR tau piav qhia saum toj no tau ua tiav tag nrho siv lub ntsuas ntsuas FAST-POCT, suav nrog lub programmed siab tswj chav tsev, chav sib nqus rho tawm, chav tswj qhov kub thiab txias, thiab fluorescent teeb liab ntes thiab ua haujlwm.Ntawm qhov kev ceeb toom, peb siv FAST-POCT platform rau RNA kev rho tawm thiab tom qab ntawd siv cov qauv RNA rho tawm rau PCR cov tshuaj tiv thaiv siv FAST-POCT system thiab desktop PCR system rau kev sib piv.Cov txiaj ntsig tau yuav luag zoo ib yam li pom hauv Daim Duab Ntxiv S8.Tus neeg teb xov tooj ua haujlwm yooj yim: qhia tus qauv rau hauv M-chamber thiab ntxig lub platform rau hauv qhov ntsuas.Cov txiaj ntsig kev ntsuas ntsuas muaj nyob hauv li 82 feeb.Cov ncauj lus kom ntxaws txog cov cuab yeej FAST-POCT tuaj yeem pom hauv daim duab ntxiv.C9, C10 thiab C11.
Influenza tshwm sim los ntawm tus kab mob khaub thuas A (IAV), B (IBV), C (ICV) thiab D (IDV) viruses yog ib qho tshwm sim thoob ntiaj teb.Ntawm cov no, IAV thiab IBV yog lub luag haujlwm rau cov neeg mob hnyav tshaj plaws thiab kev sib kis raws caij nyoog, kis 5-15% ntawm cov neeg hauv ntiaj teb, ua rau 3-5 lab tus neeg mob hnyav thiab ua rau 290,000-650,000 tus neeg tuag txhua xyoo.Cov kab mob ua pa 56,57.Kev kuaj mob ntxov ntawm IAV thiab IB yog qhov tseem ceeb los txo qis kev mob nkeeg thiab cuam tshuam txog kev lag luam.Ntawm cov txheej txheem kuaj mob muaj, qhov rov qab transcriptase polymerase saw cov tshuaj tiv thaiv (RT-PCR) yog suav tias yog qhov tseem ceeb tshaj plaws, tshwj xeeb, thiab raug (> 99%) 58,59.Ntawm cov txheej txheem kuaj mob muaj, qhov rov qab transcriptase polymerase saw cov tshuaj tiv thaiv (RT-PCR) yog suav tias yog qhov tseem ceeb tshaj plaws, tshwj xeeb, thiab raug (> 99%) 58,59.Среди доступных диагностических методов полимеразная цепная реакция с обратной транскрчиптабтей (яТ- увствительной, специфичной и точной (> 99%) 58,59.Ntawm cov txheej txheem kuaj mob muaj, rov qab transcriptase polymerase saw cov tshuaj tiv thaiv (RT-PCR) yog suav tias yog qhov tseem ceeb tshaj plaws, tshwj xeeb thiab raug (> 99%) 58,59. Из доступных диагностических методов полимеразная цепная реакция с обратной транскриптазой (Овступная реакция транскриптазой) твительной, специфичной и точной (> 99%) 58,59.Ntawm cov txheej txheem kuaj mob muaj, rov qab transcriptase polymerase saw cov tshuaj tiv thaiv (RT-PCR) yog suav tias yog qhov tseem ceeb tshaj plaws, tshwj xeeb thiab raug (> 99%) 58,59.Txawm li cas los xij, cov txheej txheem RT-PCR ib txwm xav tau rov ua cov raj xa dej, sib tov, xa tawm thiab hloov cov kua dej, txwv tsis pub lawv siv los ntawm cov kws tshaj lij hauv cov chaw muaj kev txwv.Ntawm no, FAST-POCT platform tau siv rau PCR nrhiav pom ntawm IAV thiab IBV, feem, kom tau txais lawv qhov kev txwv qis ntawm kev kuaj pom (LOD).Tsis tas li ntawd, IAV thiab IBV tau muab faib ua ntau hom kev sib cais ntawm cov kab mob sib txawv thoob plaws txhua hom, muab lub platform cog lus rau kev txheeb xyuas caj ces thiab muaj peev xwm kho tus kab mob kom raug.
Ntawm daim duab.5a qhia cov txiaj ntsig ntawm HAV PCR kuaj siv 150 µl ntawm purified viral RNA ua piv txwv.Ntawm daim duab.5a (i) qhia tau hais tias ntawm HAV concentration ntawm 106 daim ntawv / ml, fluorescence siv (ΔRn) tuaj yeem ncav cuag 0.830, thiab thaum lub concentration txo mus rau 102 daim / ml, ΔRn tseem tuaj yeem ncav cuag 0.365, uas sib haum mus rau siab dua qhov ntawd. ntawm pawg tswj tsis zoo (0.002), kwv yees li 100 npaug siab dua.Rau kev ua kom muaj nuj nqis raws li rau 6 qhov kev sim ywj pheej, ib qho kev sib tw linear calibration tau tsim los ntawm cov log concentration thiab lub voj voog qhov pib (Ct) ntawm IAV (Fig. 5a(ii)), R2 = 0.993, xws li 102-106 luam / mL.Cov txiaj ntsig tau pom zoo nrog cov txheej txheem RT-PCR.Ntawm daim duab.5a(iii) qhia cov duab fluorescent ntawm cov txiaj ntsig kev xeem tom qab 40 lub voj voog ntawm FAST-POCT platform.Peb pom tias FAST-POCT platform tuaj yeem ntes HAV tsawg li 102 daim ntawv / mL.Txawm li cas los xij, cov txheej txheem ib txwm muaj tsis muaj Ct tus nqi ntawm 102 daim ntawv / mL, ua rau LOD txog 103 daim / mL.Peb xav tias qhov no yuav yog vim qhov ua tau zoo ntawm kev sib xyaw ua npuas.PCR xeem thwmsim tau ua nyob rau hauv purified IAV RNA los soj ntsuam ntau txoj kev sib tov, nrog rau co mixing (tib yam mixing method raws li nyob rau hauv cov pa RT-PCR lag luam), vial mixing (txoj kev no, 3 s ntawm 0.12 bar) thiab tsis muaj mixing raws li ib tug tswj pawg. ..Cov txiaj ntsig tuaj yeem pom hauv Daim Duab Ntxiv S12.Nws tuaj yeem pom tau tias ntawm qhov siab dua RNA concentration (106 daim ntawv luam / mL), Ct qhov tseem ceeb ntawm cov kev sib xyaw sib txawv yuav luag zoo ib yam li kev sib xyaw ua npuas.Thaum RNA concentration poob mus rau 102 daim ntawv / mL, qhov sib tov sib xyaw thiab kev tswj tsis muaj Ct qhov tseem ceeb, thaum txoj kev sib xyaw ua npuas tseem muab tus nqi Ct ntawm 36.9, uas yog qis dua Ct qhov pib ntawm 38. Cov txiaj ntsig tau qhia tias muaj kev sib xyaw ua ke tseem ceeb. vesicles, uas kuj tau pom nyob rau hauv lwm yam ntaub ntawv, uas tej zaum kuj piav qhia yog vim li cas qhov rhiab heev ntawm FAST-POCT platform yog me ntsis siab dua li cov pa RT-PCR.Ntawm daim duab.5b qhia cov txiaj ntsig ntawm PCR tsom xam ntawm purified IBV RNA cov qauv xws li 101 txog 106 daim ntawv / ml.Cov txiaj ntsig tau zoo ib yam li IAV test, ua tiav R2 = 0.994 thiab LOD ntawm 102 daim ntawv / mL.
PCR tsom xam ntawm tus kab mob khaub thuas A (IAV) nrog IAV concentrations xws li 106 mus rau 101 daim ntawv / mL siv TE buffer raws li kev tswj tsis zoo (NC).(i) Real time fluorescence nkhaus.(ii) Linear calibration curve between logarithmic IAV RNA concentration and cycle threshold (Ct) for Fast and pa test method.(iii) IAV FAST-POCT fluorescent image after 40 cycles.b, PCR kuaj pom tus kab mob khaub thuas B (IBV) nrog (i) fluorescence spectrum real-time.(ii) Linear calibration curve thiab (iii) FAST-POCT IBV fluorescence image tom qab 40 cycles.Qhov kev txwv qis ntawm kev kuaj pom (LOD) rau IAV thiab IBV siv FAST-POCT platform yog 102 daim ntawv luam / mL, uas yog qis dua li cov qauv siv (103 luam / mL).c Multiplex test results for IAV and IBV.GAPDH tau siv los ua kev tswj xyuas qhov zoo thiab TE tsis yog siv los ua kev tswj tsis zoo los tiv thaiv kev kis kab mob thiab kev nthuav dav tom qab.Plaub hom qauv sib txawv tuaj yeem paub qhov txawv: (1) GAPDH-tsuas yog cov qauv tsis zoo (“IAV-/IBV-”);(2) IAV infection (“IAV+/IBV-”) with IAV and GAPDH;(3) IBV infection (“IAV-/IBV+”) with IBV and GAPDH;(4) IAV/IBV infection (“IAV+/IBV+”) with IAV, IBV and GAPDH.Cov kab dotted sawv cev rau txoj kab pib.n = 6 kev sim tshuaj lom neeg ywj pheej tau ua, cov ntaub ntawv tau pom tias yog ± tus qauv sib txawv.Cov ntaub ntawv nyoos yog nthuav tawm raws li cov ntaub ntawv raw cov ntaub ntawv.
Ntawm daim duab.5c qhia cov txiaj ntsig ntawm multiplexing test rau IAV/IBV.Ntawm no, tus kab mob lysate tau siv los ua cov qauv kev daws teeb meem nyob rau hauv qhov chaw ntawm purified RNA, thiab plaub primers rau IAV, IBV, GAPDH (zoo tswj) thiab TE tsis (kev tswj tsis zoo) tau ntxiv rau plaub qhov sib txawv cov tshuaj tiv thaiv ntawm FAST-POCT platform.Kev tswj qhov zoo thiab tsis zoo yog siv ntawm no los tiv thaiv kev kis kab mob thiab txhim kho keeb kwm yav dhau los.Cov kev ntsuam xyuas tau muab faib ua plaub pawg: (1) GAPDH-negative samples (“IAV-/IBV-”);(2) IAV-infected (“IAV+/IBV-”) versus IAV and GAPDH;(3) IB-.kis (“IAV-”) -/IBV+”) IBV and GAPDH;(4) IAV/IBV (“IAV+/IBV+”) infection with IAV, IBV and GAPDH.Ntawm daim duab.5c qhia tau hais tias thaum siv cov qauv tsis zoo, qhov fluorescence siv ΔRn ntawm qhov zoo tswj chamber yog 0.860, thiab ΔRn ntawm IAV thiab IBV zoo ib yam li kev tswj tsis zoo (0.002).Rau IAV +/IBV-, IAV-/IBV+ thiab IAV+/IBV+ pawg, IAV/GAPDH, IBV/GAPDH thiab IAV/IBV/GAPDH cov koob yees duab pom cov fluorescence tseem ceeb, raws li, thaum lwm lub koob yees duab txawm pom fluorescence siv ntawm keeb kwm yav dhau. theem ntawm 40 tom qab thermal cycling.Los ntawm cov kev ntsuam xyuas saum toj no, FAST-POCT platform tau qhia qhov tshwj xeeb thiab tso cai rau peb mus rau tib lub sijhawm tus kab mob khaub thuas sib txawv.
Txhawm rau txheeb xyuas qhov kev siv tau ntawm FAST-POCT, peb tau sim 36 qhov chaw kho mob (qhov ntswg swab specimens) los ntawm cov neeg mob IB (n=18) thiab cov tswj tsis-IB (n=18) (Daim duab 6a).Cov ntaub ntawv tus neeg mob tau nthuav tawm hauv Cov Lus Ntxiv 3. Cov xwm txheej ntawm tus kab mob IB tau lees paub ntawm nws tus kheej thiab cov txheej txheem kawm tau pom zoo los ntawm Zhejiang University First Affiliated Hospital (Hangzhou, Zhejiang).Txhua tus qauv ntawm cov neeg mob tau muab faib ua ob pawg.Ib qho tau ua tiav siv FAST-POCT thiab lwm qhov tau ua tiav siv lub desktop PCR system (SLAN-96P, Suav).Ob qhov kev ntsuam xyuas siv tib yam khoom siv kom huv thiab kuaj pom.Ntawm daim duab.6b qhia cov txiaj ntsig ntawm FAST-POCT thiab cov lus qhia rov qab rov qab PCR (RT-PCR).Peb piv cov fluorescence siv (FAST-POCT) nrog -log2 (Ct), qhov twg Ct yog lub voj voog pib rau cov pa RT-PCR.Muaj kev pom zoo zoo ntawm ob txoj hauv kev.FAST-POCT thiab RT-PCR pom muaj kev sib raug zoo nrog Pearson tus piv (r) tus nqi ntawm 0.90 (Daim duab 6b).Peb mam li ntsuam xyuas qhov tseeb ntawm FAST-POCT.Fluorescence intensity (FL) distributions for positive and negative samples tau muab los ua ib qho kev ntsuas ywj pheej (Fig. 6c).FL qhov tseem ceeb tau nce siab dua hauv cov neeg mob IB dua li kev tswj hwm (****P = 3.31 × 10-19; ob-tailed t-test) (Fig. 6d).Tom ntej no, IBV receiver operating yam ntxwv (ROC) nkhaus tau npaj.Peb pom tias qhov tseeb ntawm kev kuaj mob tau zoo heev, nrog thaj tsam hauv qab ntawm qhov nkhaus ntawm 1 (Fig. 6e).Thov nco ntsoov tias vim qhov yuav tsum tau ua daim npog ntsej muag hauv Suav teb vim yog COVID-19 txij li xyoo 2020, peb tsis tau txheeb xyuas cov neeg mob IBD, yog li txhua qhov kev kuaj mob zoo (xws li, qhov ntswg qhov ntswg) yog rau IBV nkaus xwb.
Kev tsim kho kev kawm.Tag nrho ntawm 36 tus qauv, suav nrog 18 tus neeg mob kuaj thiab 18 qhov kev tswj tsis tau mob khaub thuas, tau txheeb xyuas siv FAST-POCT platform thiab cov qauv RT-PCR.b Txheeb xyuas qhov sib xws ntawm FAST-POCT PCR thiab cov qauv RT-PCR.Cov txiaj ntsig tau muaj txiaj ntsig zoo (Pearson r = 0.90).c Fluorescence siv theem hauv 18 tus neeg mob IB thiab 18 tswj.d Hauv cov neeg mob IB (+), FL qhov tseem ceeb tau nce siab dua hauv pawg tswj hwm (-) (****P = 3.31 × 10-19; ob-tailed t-test; n = 36).Rau txhua daim duab square, tus cim dub nyob rau hauv nruab nrab sawv cev rau qhov nruab nrab, thiab cov kab hauv qab thiab sab saum toj ntawm lub thawv sawv cev rau 25th thiab 75th feem pua, feem.Cov whiskers txuas mus rau qhov tsawg kawg nkaus thiab siab tshaj cov ntaub ntawv cov ntsiab lus, uas tsis suav tias yog outliers.e ROC nkhaus.Cov kab dotted d sawv cev rau tus nqi pib kwv yees los ntawm ROC tsom xam.AUC rau IBV yog 1. Cov ntaub ntawv nyoos yog muab ua cov ntaub ntawv nyoos.
Hauv tsab xov xwm no, peb nthuav qhia FAST, uas muaj cov yam ntxwv xav tau rau POCT zoo tagnrho.Qhov zoo ntawm peb cov thev naus laus zis muaj xws li: (1) Kev siv ntau yam (cascade, ib txhij, ua ntu zus thiab xaiv), tso tawm ntawm qhov kev thov (kev tso tawm sai thiab sib npaug ntawm kev thov siab) thiab kev ua haujlwm ruaj khov (kev vibration ntawm 150 degrees) (2) kev cia ntev (2 xyoos ntawm kev ntsuas nrawm, poob qis txog 0.3%);(3) muaj peev xwm ua hauj lwm nrog cov kua dej uas muaj ntau yam wettability thiab viscosity (viscosity txog 5500 cP);(4) Kev Lag Luam (Kev kwv yees tus nqi ntawm cov khoom siv FAST-POCT PCR yog kwv yees li US $ 1).Los ntawm kev sib txuas cov khoom siv ntau yam, kev sib koom ua ke FAST-POCT platform rau PCR tshawb pom tus kab mob khaub thuas A thiab B tau pom thiab siv.FAST-POCT tsuas yog siv sijhawm 82 feeb xwb.Cov kev kuaj mob nrog 36 qhov ntswg swab kuaj tau pom tias muaj kev sib raug zoo hauv kev siv fluorescence nrog tus qauv RT-PCR (Pearson coefficients> 0.9).Cov kev kuaj mob nrog 36 qhov ntswg swab kuaj tau pom tias muaj kev sib raug zoo hauv kev siv fluorescence nrog tus qauv RT-PCR (Pearson coefficients> 0.9).Клинические тесты с 36 образцами мазков из носа показали хорошее сответствие интенсивности флуойцас Р (коэффициенты Пирсона > 0,9).Kev kuaj mob nrog 36 cov qauv ntawm qhov ntswg qhov ntswg pom pom zoo nrog cov fluorescence siv ntawm tus qauv RT-PCR (Pearson's coefficients> 0.9).RT-PCR Клинические испытания 36 образцов мазков из носа показали хорошее совпадение интенсивности Пноноресцен и флуоресцен (khoэффициент Пирсона > 0,9).Kev kuaj mob ntawm 36 qhov ntswg qhov ntswg tau pom zoo pom zoo ntawm fluorescence siv nrog tus qauv RT-PCR (Pearson's coefficient> 0.9).Nyob rau tib lub sijhawm nrog txoj haujlwm no, ntau yam kev siv tshuaj biochemical (piv txwv li, plasma thermal cycling, amplification-free immunoassays, thiab nanobody functionalization assays) tau qhia lawv lub peev xwm hauv POCT.Txawm li cas los xij, vim tsis muaj kev sib koom ua ke thiab muaj zog POCT platform, cov txheej txheem no inevitably yuav tsum tau cais cov txheej txheem ua ntej (piv txwv li, RNA isolation44, incubation45 thiab washing46), uas ntxiv ntxiv rau kev ua haujlwm tam sim no nrog cov txheej txheem no los siv cov kev ua haujlwm siab tshaj POCT nrog. qhov yuav tsum tau muaj.fetch-in-response-output kev ua tau zoo.Hauv kev ua haujlwm no, txawm hais tias lub twj tso kua mis cua siv los ua kom lub FAST valve me me txaus los ua ke rau hauv lub rooj zaum ntsuas (Daim duab S9, S10), nws tseem siv lub zog tseem ceeb thiab ua suab nrov.Raws li txoj cai, cov twj me me pneumatic tuaj yeem hloov tau los ntawm lwm txoj kev, xws li kev siv hluav taws xob hluav taws xob lossis kev siv ntiv tes.Cov kev txhim kho ntxiv tuaj yeem suav nrog, piv txwv li, hloov cov khoom siv rau cov khoom siv tshuaj lom neeg sib txawv thiab tshwj xeeb, siv cov kev tshawb pom tshiab uas tsis tas yuav tsum muaj cua sov / cua txias, yog li muab cov cuab yeej tsis pub siv POCT platform rau PCR daim ntawv thov.Peb ntseeg hais tias muab qhov FAST platform muab ib txoj hauv kev los tswj cov kua dej, peb ntseeg tias FAST thev naus laus zis tau nthuav tawm lub peev xwm los tsim ib lub platform uas tsis yog rau kev sim tshuaj biomedical nkaus xwb, tab sis kuj tseem rau kev soj ntsuam ib puag ncig, kev soj ntsuam zaub mov zoo, khoom siv thiab tshuaj synthesis. ..
Kev sau thiab siv tib neeg lub qhov ntswg qhov ntswg tau pom zoo los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Zhejiang University First Affiliated Hospital (IIT20220330B).36 qhov ntswg swab kuaj tau raug sau, suav nrog 16 tus neeg laus <30 xyoo, 7 laus> 40 xyoo, thiab 19 txiv neej, 17 poj niam.36 qhov ntswg swab kuaj tau raug sau, suav nrog 16 tus neeg laus <30 xyoo, 7 laus> 40 xyoo, thiab 19 txiv neej, 17 poj niam.Было собрано 36 образцов мазков из носа, в которых приняли участие 16 взрослых < 30 лет, 7 в зрослы 9 в зрослы thiab 17 xyoo.Peb caug-rau qhov hnoos qeev tau sau los ntawm 16 tus neeg laus <30 xyoo, 7 tus neeg laus dua 40 xyoo, 19 tus txiv neej thiab 17 tus poj niam.Cov ntaub ntawv pej xeem tau nthuav tawm hauv Cov Lus Ntxiv 3. Cov ntaub ntawv pom zoo tau txais los ntawm txhua tus neeg koom.Txhua tus neeg koom tau raug xav tias muaj tus mob khaub thuas thiab raug kuaj los ntawm kev yeem yam tsis tau them nyiaj.
Lub hauv paus ceev thiab lub hau yog ua los ntawm polylactic acid (PLA) thiab luam tawm los ntawm Ender 3 Pro 3D tshuab luam ntawv (Shenzhen Transcend 3D Technology Co., Ltd.).Ob chav sided daim kab xev tau yuav los ntawm Adhesives Research, Inc. Model 90880. PET zaj duab xis 100 µm tuab yog yuav los ntawm McMaster-Carr.Ob lub nplaum thiab PET zaj duab xis raug txiav siv Silhouette Cameo 2 cutter los ntawm Silhouette America, Inc. Cov yeeb yaj kiab elastic yog ua los ntawm cov khoom siv PDMS los ntawm kev txhaj tshuaj molding.Ua ntej, 200 µm tuab PET thav duab raug txiav siv lub tshuab laser thiab txuas rau 3 hli tuab PMMA daim ntawv siv 100 µm ob daim kab xev nplaum.Lub PDMS precursor (Sylgard 184; Ntu A: Ntu B = 10: 1, Dow Corning) tau muab nchuav rau hauv pwm thiab siv lub iav pas nrig los tshem tawm PDMS ntau dhau.Tom qab kho ntawm 70 ° C. rau 3 teev, 300 μm tuab PDMS zaj duab xis tuaj yeem raug tshem tawm ntawm cov pwm.
Cov duab rau ntau yam kev faib tawm, kev tshaj tawm raws li qhov xav tau thiab kev ua haujlwm txhim khu kev qha yog coj nrog lub koob yees duab ceev (Sony AX700 1000 fps).Lub orbital shaker siv hauv kev ntsuas kev ntseeg tau tau yuav los ntawm SCILOGEX (SCI-O180).Cov huab cua siab yog tsim los ntawm lub tshuab cua compressor, thiab ob peb lub digital precision siab regulators yog siv los kho tus nqi siab.Txoj kev ntsuas tus cwj pwm ntws yog raws li hauv qab no.Ib qho kev txiav txim siab ntawm cov kua dej tau raug txhaj rau hauv qhov ntsuas ntsuas thiab lub koob yees duab ceev tau siv los sau tus cwj pwm ntws.Cov duab tseem tau raug coj los ntawm cov yeeb yaj kiab ntawm tus cwj pwm ntws ntawm lub sijhawm tas, thiab thaj chaw seem tau suav nrog siv Image-Pro Plus software, uas tau muab faib los ntawm lub koob yees duab qhov tob los xam qhov ntim.Cov ntsiab lus ntawm qhov kev ntsuas tus cwj pwm ntws tuaj yeem pom nyob rau hauv Daim Duab Ntxiv S4.
Txhaj 50 μl ntawm microbeads thiab 100 μl ntawm cov dej deionized rau hauv lub vial sib xyaw.Cov duab sib xyaw ua haujlwm tau muab coj los nrog lub koob yees duab ceev ceev txhua txhua 0.1 vib nas this ntawm qhov siab ntawm 0.1 bar, 0.15 bar thiab 0.2 bar.Pixel cov ntaub ntawv thaum lub sijhawm sib xyaw tuaj yeem tau txais los ntawm cov duab no siv software ua duab (Photoshop CS6).Thiab kev sib xyaw ua haujlwm tuaj yeem ua tiav nrog cov kab lus hauv qab no 53.
qhov twg M yog qhov kev sib xyaw ua haujlwm, N yog tag nrho cov qauv pixels, thiab ci thiab \(\bar{c}\) yog cov normalized thiab xav tau normalized concentrations.Kev sib xyaw ua haujlwm ntawm 0 (0%, tsis sib xyaw) mus rau 1 (100%, tag nrho sib xyaw).Cov txiaj ntsig tau pom hauv Daim Duab Ntxiv S6.
Real-time RT-PCR kit for IAV and IBV, including IAV and IBV RNA samples (cat. no. RR-0051-02/RR-0052-02, Liferiver, China), Tris-EDTA buffer (TE buffer no. B541019 , Sangon Biotech, Tuam Tshoj), Zoo Tswj RNA Cov Khoom Siv Purification (Tshooj No. Z-ME-0010, Liferiver, Tuam Tshoj) thiab GAPDH Solution (Tshooj No. M591101, Sangon Biotech, Tuam Tshoj) yog muag.Cov khoom siv RNA purification muaj xws li cov khoom siv khi, ntxuav A, ntxuav W, eluent, sib nqus microbeads, thiab cov khoom siv acrylic.IAV thiab IBV real-time RT-PCR cov khoom siv suav nrog IFVA nucleic acid PCR tshawb pom sib xyaw thiab RT-PCR enzyme.Ntxiv 6 µl ntawm AcrylCarrier thiab 20 µl ntawm cov hlaws sib nqus rau 500 µl ntawm kev sib khi tsis muaj tshuaj, co zoo thiab tom qab ntawd npaj cov hlaws daws.Ntxiv 21 ml ntawm ethanol rau ntxuav A thiab W, co zoo kom tau txais cov tshuaj ntxuav A thiab W, feem.Tom qab ntawd, 18 µl ntawm fluorescent PCR sib xyaw nrog IFVA nucleic acid thiab 1 µl ntawm RT-PCR enzyme tau ntxiv rau 1 µl ntawm TE tov, shaken thiab centrifuged rau ob peb feeb, tau 20 µl ntawm IAV thiab IBV primers.
Ua raws li cov txheej txheem RNA purification hauv qab no: (1) RNA adsorption.Pipette 526 µl ntawm cov tshuaj pellet rau hauv 1.5 ml centrifuge raj thiab ntxiv 150 µl ntawm cov qauv, ces manually co lub raj nce thiab nqis 10 zaug.Hloov 676 µl ntawm qhov sib tov mus rau affinity kem thiab centrifuge ntawm 1.88 x 104 g rau 60 vib nas this.Cov kua dej tom qab ntawd muab pov tseg.(2) Thawj theem ntawm kev ntxuav.Ntxiv 500 µl ntawm cov tshuaj ntxuav A mus rau hauv kab affinity, centrifuge ntawm 1.88 x 104 g rau 40 s, thiab pov tseg cov tshuaj siv.Txoj kev ntxuav no tau rov ua dua ob zaug.(3) theem ob ntawm kev ntxuav.Ntxiv 500 µl ntawm cov tshuaj ntxuav W mus rau kab affinity, centrifuge ntawm 1.88 × 104 g rau 15 s thiab pov tseg cov tshuaj siv.Txoj kev ntxuav no tau rov ua dua ob zaug.(4) Kev ntseeg.Ntxiv 200 µl ntawm eluate rau affinity kem thiab centrifuge ntawm 1.88 x 104 g rau 2 min.(5) RT-PCR: Cov eluate tau txhaj rau hauv 20 μl ntawm cov tshuaj primer hauv PCR raj, tom qab ntawd lub raj tau muab tso rau hauv lub sijhawm tiag tiag PCR kuaj apparatus (SLAN-96P) kom ua tiav cov txheej txheem RT-PCR.Tag nrho cov txheej txheem tshawb pom yuav siv li ntawm 140 feeb (20 feeb rau RNA purification thiab 120 feeb rau PCR nrhiav).
526 µl ntawm cov hlaws dai, 1000 µl ntawm cov tshuaj ntxuav A, 1000 µl ntawm cov tshuaj ntxuav W, 200 µl ntawm eluate thiab 20 µl ntawm primer tov tau muab tso ua ntej thiab khaws cia hauv chav M, W1, W2, E thiab PCR nrhiav kom pom.Platform sib dhos.Tom qab ntawd, 150 µl ntawm cov qauv tau muab tso rau hauv chamber M thiab FAST-POCT platform tau muab tso rau hauv qhov ntsuas ntsuas tau pom hauv Daim Duab Ntxiv S9.Tom qab kwv yees li 82 feeb, qhov kev xeem tau muaj.
Tshwj tsis yog tias tau sau tseg, tag nrho cov txiaj ntsig kev xeem tau nthuav tawm raws li qhov txhais tau tias ± SD tom qab tsawg kawg ntawm rau qhov rov ua dua siv tsuas yog FAST-POCT platform thiab cov qauv kev ywj pheej lom neeg.Tsis muaj cov ntaub ntawv raug tshem tawm los ntawm kev txheeb xyuas.Cov kev sim tsis yog random.Cov kws tshawb fawb tsis tau dig muag rau pab pawg ua haujlwm thaum lub sijhawm sim.
Yog xav paub ntxiv txog kev tsim qauv, saib Nature Research Report abstract linked to this article.
Cov ntaub ntawv txhawb nqa cov txiaj ntsig ntawm txoj kev tshawb fawb no muaj nyob hauv Cov Lus Qhia Ntxiv.Kab lus no muab cov ntaub ntawv qub.
Chagla, Z. & Madhukar, P. COVID-19 txhawb nqa hauv cov tebchaws nplua nuj yuav ncua sijhawm txhaj tshuaj tiv thaiv rau txhua tus.Chagla, Z. & Madhukar, P. COVID-19 txhawb nqa hauv cov tebchaws nplua nuj yuav ncua sijhawm txhaj tshuaj tiv thaiv rau txhua tus.Chagla, Z. thiab Madhukar, P. COVID-19 txhawb nqa hauv cov tebchaws nplua nuj yuav ncua sijhawm txhaj tshuaj tiv thaiv rau txhua tus.Chagla, Z. thiab Madhukar, P. COVID-19 revaccination nyob rau hauv cov teb chaws nplua nuj yuav ncua kev txhaj tshuaj tiv thaiv rau txhua tus.National tshuaj.27, 1659–1665 (2021).
Faust, L. et al.SARS-CoV-2 testing in low- and medium- income countries: available and affordability in the private healthcare sector.kab mob microbial.22, 511–514 (2020).
Koom Haum Saib Xyuas Kev Noj Qab Haus Huv Thoob Ntiaj Teb.Kev nthuav dav thoob ntiaj teb thiab qhov tshwm sim ntawm kev xaiv cov kab mob sib kis tau yooj yim: kev tshuaj xyuas thiab kwv yees.Geneva: WHO, WHO/HIV_AIDS/2 https://apps.who.int/iris/bitstream/handle/10665/66818/WHO_HIV_AIDS_2001.02.pdf (2001).
Fenton, EM et al.Ntau 2D molded sab txaus kuaj strips.ASS daim ntawv thov.alma mav.Inter Milan.1, 124–129 (2009).
Schilling, KM et al.Tag nrho cov khoom siv microfluidic daim ntawv tsom xam.qhov quav.Tshuaj.84, 1579–1585 (2012).
Lapenter, N. et al.Competitive paper-based immunochromatography ua ke nrog enzyme-hloov electrodes tso cai rau kev saib xyuas wireless thiab kev txiav txim siab electrochemical ntawm cov zis cotinine.Sensors 21, 1659 (2021).
Zhu, X. et al.Kev ntsuas tus kab mob biomarkers nrog ntau yam nanozyme-kev sib xyaw ua ke ntawm cov kua dej sib txuas siv lub glucometer.biological sensor.Bioelectronics.126, 690–696 (2019).
Boo, S. et al.Kev kuaj cev xeeb tub rau kev kuaj xyuas cov kab mob pathogenic siv concanavalin A-tib neeg chorionic gonadotropin-Cu3 (PO4)2 hybrid nanoflowers, sib nqus sib nqus thiab nyeem xov tooj ntse.Microcomputer.Phau ntawv xov xwm.185, 464 (2018).