Hielscher faka'ata tekinolosia

Protocol ki he SARS-KoloV-2 Coronavirus fakamalohia mo Sonication

Ko e Hielscher VialTweeter ko ha ʻiuniti makehe ultrasonic fakavahaʻa-taʻu, ʻa ia ʻoku fakaʻaongaʻi ke inactivate ʻa e coronavirus SARS-KOV-2. ʻOku fakaʻata ʻe he VialTweeter ke teuteu ki ha sipinga ʻe 10 ʻi he taimi tatau pe pea ʻoku ʻi ai ʻa e ʻiuniti lelei taha ki he sipinga ngaue lahi.

Fakamalohia ʻo e SARS-KoloV-2 Coronavirus mo e VialTweeter

Hili hono toʻo e monolayers, naʻe tuʻo tolu hono fufulu mo phosphate buffered Saline (PBS) kimuʻa pea toki maʻu e ngaahi selo ki he 1mL MEM/5% FBS mo sonicated (3 × 10 ʻi he, sekoni ʻe 10 mei he 100% malohi mo e amplitude) ʻo fakaʻaongaʻi ʻa e processor Hielscher UP200St mo VialTweeter pipiki ki ai. Naʻe fakamahinoʻi Supernatants ʻe centrifuging ʻi he 3000 × g ʻi ha miniti ʻe 10. 

Lau kakato ʻa e ngaahi founga fakangaue ʻi heni maʻa e SARS-KoloV-2 coronavirus fakamalohia ʻe Welch et Al. (2020) ʻi lalo:

Ngaahi selo mo e vailasi

Vero E6 selo (Vero C1008; ATCC CRL-1586) ʻi hono fakaleleiʻi e founga mahuʻinga siʻisiʻi taha ʻo e ʻIkale (MEM) fakalahi ki he 10% (v/v) fetal ʻuhikiʻi pulu serum (CS). Ko e vailasi naʻe fakaʻaongaʻi ko e SARS-KoloV-2 faingataʻaʻia hCOV-19/ʻIngilani/2/2020, mamaʻo mei he PHE mei he fuofua kau mahaki ʻi he UK ʻi he 29/01/2020. Naʻe maʻu ʻa e vailasi ko ʻeni ʻi he potufolofola 1 pea fakaʻaongaʻi ia ki he ako fakamalohia ʻi he potufolofola 2 pe 3.
Ki he reagents mo e ngaahi kemikale ʻoku fakaʻaongaʻi ki he SARS-KoloV-2 pea pehe ki hono toʻo ʻo e reagent cytotoxicity kataki ʻo sio ki he lipooti fakasaienisi ʻo welch et Al. (2020).

The ultrasonic sample preparation unit VialTweeter is used for the inactivation of SARS-CoV-2 coronavirus

Fakamalohia ʻo e vailasi ʻe he koa fufulu koa – SARS-KoloV-2 Coronavirus fakamalohia Protocol ʻe Welch et Al. 2020

SARS-KoloV-2 fakamalohia

Ki he ngaahi naunau fakakomesiale, teuteu ki he vailasi (lolo moluu, titers kamata mei he 1 × 106 ki he 1 × 108 PfU/ml) ʻi he triplicate mo e reagents ʻi he concentrations pea mo e taimi fetuʻutaki ʻoku fokotuʻu atu ʻi he ngaahi fakahinohino ki hono fakaʻaongaʻi, feituʻu ʻoku lava ke maʻu, pe ki he concentrations mo e taimi pau ʻoku kole ʻe he sivi laboratories. ʻI he feituʻu naʻe fai ai ha concentrations ʻe he kautaha ngaohi kapeti, naʻe siviʻi ai ʻa e tuʻunga maʻulalo taha ʻo e koloa ki he sipinga (ʻa ia ko e maʻulalo taha ia ʻoku fokotuʻu atu ki hono siviʻi ʻo e koloa). Naʻe siviʻi Specimen foʻi tiupi reagents ʻo fakaʻaongaʻi ha tohi ʻe taha ʻo e ngaahi ʻulungaanga fakafonua ʻe hongofulu ki he reagent, tuku kehe kapau naʻe fakamahinoʻi mai ʻe he kautaha naʻa nau ngaohi ha lolo ke reagent. Naʻe siviʻi e koa fufulu fakamaʻa, fixatives mo e solvents ʻi he concentrations ʻoku fakaha atu ʻi he taimi ʻoku fakaʻilongaʻi atu. Naʻe fakahoko kotoa e ngaahi sitepu ki hono fakamalohia ʻo e loki (18 – 25 ° c). Ke siviʻi e ngaahi faʻahinga sipinga ʻe taha, naʻe heʻeku ʻa e vailasi ki he sipinga ʻoku fakaʻilongaʻi matrix ʻi ha fika (ratio) ʻo e 1:9, pea fai ha sivi reagents ʻo hange ko ia ʻi ʻolunga. Naʻe kau ʻi he ngaahi fakatotolo ʻahiʻahi kotoa pe ʻa hono puleʻi ʻo e ngaahi sipinga ʻoku manukiʻi ʻaki ʻa e PBS ʻo fetongi ʻaki ʻa e sivi reagent. Hili pe ʻa e taimi fetuʻutaki ʻoku fie maʻu, naʻe ngaue ʻaki ʻa e 1mL ʻo e sipinga ʻo e ngaahi meʻa naʻe fili totonu matrix. Naʻe fakahoko Reagent hono toʻo ʻo e sivi ʻo e fakamalohia ʻi ha kolomu lahi ange ʻo fakaʻaongaʻi ʻa e Piesa 4mL fo toʻo e ʻu kolomu (fakapaima toutai), pe ko hono fakafonu ʻo e ngaahi kolomu ʻoku ʻikai ke ʻi ai ha meʻa centrifuge (Fakapaatomi toutai) mo SM2 piokalafi-ʻasi, Sephacryl S-400HR pe Sephadex LH-20 ke ʻoange 4mL ne ʻasi/nofoʻanga. Ke fakamaʻa ʻaki hono fakaʻaongaʻi ʻo e Amicon meʻasivi, 2 × 500μl naʻe fakamaʻa ʻa e ngaahi sipinga ʻo fakaʻaongaʻi ha meʻasivi centrifugal ʻe ua ʻi he founga ne fakamatalaʻi kimuʻa, pea toki fakatahaʻi. Ke formaldehyde mo formaldehyde ʻaki glutaraldehyde toʻo, naʻe fakaʻaongaʻi ha sivi ʻe taha ʻaki ha sipinga ʻe 1× 500μl, resuspended hili e ngaue ʻi he 500μl PBS, pea tanaki atu ki he 400ul MEM/5% FBS. Ki hono fakamalohia ʻo e mahaki VialTweeter at the ultrasonic processor UP200STmonolayers, 12.5 cm2 flasks ʻo Vero E6 ngaahi selo (2.5 × 106 naʻe uesia ʻa e ngaahi selo/flask ʻi he 2.5 mL MEM/5% FBS) ʻi he MOI 0.001 pea incubated ʻi he 37 ° c/5% CO2 ʻi ha houa ʻe 24. Naʻe toʻo Supernatant, pea fakaleleiʻi ʻa e ngaahi selo ʻo fakaʻaongaʻi ʻa e 5mL ʻo formaldehyde, pe formaldehyde mo glutaraldehyde ʻi he mafana ʻo e loki ʻi ha miniti ʻe 15 pe 60. Naʻe toʻo ʻa e fixative, pea monolayers tuʻo tolu ʻaki ʻa e PBS kimuʻa pea toki maʻu ha ngaahi selo ki 1mL MEM/5% FBS mo sonicated (3 × ua ʻi he, sekoni ʻe 10 ʻi he 100% malohi mo e amplitude) ʻo fakaʻaongaʻi ha UP200St mo VialTweeterter (Hielssoun tekinolosia). Naʻe fakamahinoʻi Supernatants ʻe centrifuging ʻi he 3000 × g ʻi ha miniti ʻe 10.

VialTweeter ki he Sonication lahi 'o e Vials kuo tapuni

VialTweeter ki he Sonication lahi ʻo e ngaahi meʻa kuo tapuni

Kole 'a e fakamatala




Fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


Various reagents have been tested for the inactivation of the SARS-CoV-2 coronavirus using the ultrasonic sample prep unit VialTweeter.

Fakaikiiki ʻo e reagent ʻoku fakaʻaongaʻi ki he SARS-KoloV-2 coronavirus fakamalohia mo e ultrasonic VialTweeter ʻo fakatatau ki he ngaahi founga fakangaue ʻa Welch et Al. 2020

ʻE lava ke maʻu heni ʻa e ngaahi founga kakato kau ai hono fakaʻaongaʻi ʻo e Hielscher VialTweeter:
Welch, Stephen R.; Davies, Katherine A.; Buczkowski, Hubert; Hettiarachchi, Nipunadi; Green, Nicole; Arnold, Ulrike; Jones, Matthew; Hannah, Matthew J.; Evans, Reah; Burton, Christopher; Burton, Jane E.; Guiver, Malcolm; Cane, Patricia A.; Woodford, Neil; Bruce, Christine B.; Roberts, Allen D. G.; Killip, Marian J. (2020): Inactivation analysis of SARS-CoV-2 by specimen transport media, nucleic acid extraction reagents, detergents and fixatives. Journal of Clinical Microbiology. Accepted Manuscript Posted Online 24 August 2020.

The multi-sample ultrasonicator VialTweeter allows for simultaneous sample preparation of up to 10 vials under same process conditions. The VialTweeter is an established ultrasonic device used for the inactivation of corona virus SARS-CoV-2 (Click to enlarge!)

VialTweeter ki ultrasonic fakamalohia ʻo e vailasi

Ngaahi lelei 'o e VialTweeter ha vakai nounou ki

  • Sonication ʻo aʻu ki he 10 ʻi he taimi tatau pe
  • ʻIkai ha fakaʻuliʻi ʻi he kolosi
  • ʻIkai ha sipinga ʻo e mole
  • Lekooti ʻo e fakamatala fakahangatonu
  • Faingofua mo malu ke ngaue
ʻOku toe fakaʻaongaʻi foki ʻa e VialTweeter ki he

  • To'oto'o Lysis
  • Viral fakahohaʻasi e konga siʻi
  • Nucleic e toʻo ʻuli ʻa e ʻesiti: DNA/RNA fakamavaheʻi
  • DNA/RNA fragmentation
  • Lysate solubilization
  • Fakaʻata Dismembrators mo e Disruptors toʻotoʻo

    Ko e sipinga fakavahaʻa-ultrasonicator VialTweeter ko e taha pe ia ʻo e ngaahi founga ultrasonic lahi ki hono teuteuʻi totonu, biochemical mo kiliniki laboratories. ʻOku ʻomi ʻe Hielscher Ultrasonics ʻa e lelei taha ultrasonic dismembrator ki hoʻo tohi kole, hange ko e lysis toʻotoʻo, toʻo hingoa toʻotoʻo, homogenization, lysate solubilization, fakaleleiʻi, sipinga degassing etc.
    Tau ʻilo muʻa pe ko e sipinga ʻe fiha kuo pau ke ke ngaue ki ai he houa mo e ʻaho, kapau ʻoku ke saiʻia ʻi he sonication fakahangatonu pe ʻikai fakahangatonu pea mo e taumuʻa ʻo e ultrasonic sipinga ʻo e faitoʻo. Te mau fokotuʻu atu ʻa e ʻiuniti ultrasonic feʻunga taha ki hoʻo ngaue fakaʻaho!
    Hielscher Ultrasonics ʻ Digital ultrasonic processors kuo fakanaunau ʻaki ia ʻa e polokalama fakakomipiuta, lekooti fakahangatonu ʻo e fakamatala, ngaahi fili faingofua ki hono mapuleʻi ʻo e ʻea, taimi sonication, siakale/sepo pea pehe ki he sipinga ʻo e maama mo e browser ʻoku mamaʻo hono puleʻi. ʻOku tau feinga ke ʻai ʻetau ngaahi meʻangaue ultrasonic ki he poto taha ʻe lava koeʻuhi ke lava hoʻo fakatotolo mo e ngaue angamaheni ʻo faingamalie mo lavameʻa ʻi he lahi taha ʻe lava.
    Lomiʻi heni, ke maʻu ha ngaahi founga lahi ange kau ai ʻa e ultrasonic sipinga ʻo e teuteu mo e VialTweeter!

    Fetu'utaki mai kiate kimautolu! / Kole kiate kitautolu!

    Kole ki ha fakamatala lahi ange

    Kataki 'o ngaue 'aki e foomu 'i lalo ke kole ha toe fakamatala ki he ultrasonic processors, faka'aonga'i mo e totongi. Te mau fiefia ke alea'i e founga 'oku mo 'i ai mo ho'omou ngaahi fie ma'u ultrasonic he polokalama!









    Kataki 'o fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


    Ultrasonic high-shear homogenizers are used in lab, bench-top, pilot and industrial processing.

    Hielscher Ultrasonics ʻa e ngaohiʻanga koloa maʻolunga ultrasonic homogenizers ki hono tuifio ʻo e ngaahi polokalama, fakamovetevetea, emulsification mo e toʻo hingoa ʻi he fale fakatotolo fakakemi, pailate mo e meʻafua ki he ngaahi meʻa fakapisinisi.

    Ngaahi tohi/fakamo'oni fakafolofola



    Ngaahi mo'oni'i me'a 'oku mahu'inga ke 'ilo'i

    Ko e ha ʻa e ngaahi selo Vero?

    Vero E6, naʻe toe ʻiloa foki ko Vero C1008 (ATCC fika Ko e CRL-1586) ko e laine toʻotoʻo pe ia mei Vero 76 pea ʻoku fakaʻaongaʻi ia ʻi he fekumi ʻa SARS-KoloV mo e SARS-KoloV-2 coronaviruses. Ko e ngaahi selo Vero ko ha hako ia ʻoku fakaʻaongaʻi ʻi he ngaahi anga fakafonua toʻotoʻo. Ko e ʻ Vero’ Naʻe mamaʻo ʻa e hako mei he kofuua epithelial ngaahi selo naʻe toʻo mei ha ngeli lanumata ʻi ʻAfilika (Chlorocebus SP.).
    ʻOku fakahaaʻi ʻe Vero E6 ngaahi selo ha fetuʻutaki inhibition, ko ia ʻoku feʻunga mo propagating vailasi ʻoku ako mamalie. ʻOku angamaheni ʻaki hono fakaʻaongaʻi ʻa e Vero E6 laine toʻotoʻo ke vakaiʻi ʻaki ʻa e cytopathology ʻo coronaviruses SARS-KoloV mo e SARS-Kov-2 ko ha ngaahi selo Vero (ngaahi selo ʻo e kofuua lanumata) ʻoku nau fakahaaʻi ha fakafotunga lahi ʻo e angiotensin-fakaului enzyme 2 (ACE2) meʻa puke. Ko e ACE2 meʻa puke ko ha feituʻu lahi ia ʻo e tuʻi ki he SARS-Kov-2 coronavirus.
    Hange ko ʻeni, naʻe ʻilo ʻe Ogando et Al. (2020) ko e SARS-KoloV-2 – ʻi hono fakafehoanaki ki he SARS-KoloV – ha ngaahi tuʻunga maʻolunga ange ʻo e intracellular viral RNA, ka naʻe siʻisiʻi ange ʻa e takanga ʻa e 50 viral progeny mei he anga fakafonua. ʻIkai ngata ai, naʻa na fakapapauʻi ko e ongoʻingofua ko ia ʻo e ongo vailasi ki ha inhibitors ʻe tolu kuo fokotuʻu inhibitors ʻo coronavirus replication (Remdesivir, Alisporivir mo chloroquine) ʻoku meimei ke fuʻu tokanga ʻaupito ki hono faitoʻo ʻo e ngaahi selo ʻaki pegylated ʻAlifa. Ko ha faikehekehe mahuʻinga ʻi he ongo vailasi ko e foʻi moʻoni ko ia – ʻi passaging ʻi vero ngaahi selo E6 – ʻOku mahino ko e SARS-KoloV-2 ʻoku malohi hono fili ke maʻu adaptive miuteisoni ʻi hono ivi spike Sini. ʻOku hanga ʻe he miuteisoni ko ʻeni ʻo liliu pe tamateʻi ha putative ʻ furin hange ko cleavage ʻ ʻi he vahefonua ʻoku ne fakafehokotaki ʻa e S1 mo e S2 domains pea ko hono ola ko ha liliu ʻiloa phenotypic ʻi he tatongitongi ʻo e tatongitongi.