I-Hielscher Ultrasound Technology

Iphrothokholi ye-SARS-CoV-2 Coronavirus Inactivation ne-Sonication

I-Hielscher VialTweeter iyunithi eyingqayizivele yokulungiselela amasampula amaningi we-ultrasonic, esetshenziselwa ukungasebenzi i-coronavirus SARS-COV-2. I-VialTweeter ivumela ukulungiselela izigaxa zesampula ezifika ku-10 ngasikhathi sinye futhi ngaleyo ndlela iyunithi ekahle yokucubungula amasampula amaningi.

Ukungasebenzi kwe-SARS-CoV-2 Coronavirus nge-VialTweeter

Ngemuva kokususa i-fixative, ama-monolayers ahlanzwa kathathu nge-phosphate-buffered saline (PBS) ngaphambi kokukhuhla amaseli abe yi-1mL MEM / 5% FBS futhi enziwe amandla (3 × 10 wesibili, amasekhondi ayi-10 ku-100% amandla ne-amplitude) kusetshenziswa i-Hielscher iprosesa le-ultrasonic UP200St ngeVialTweeter okunamathiselwe. Ama-supernatants acacisiwe ngokufakwa kwe-centrifuging ku-3000 × g imizuzu eyi-10. Funda umthetho olandelwayo ogcwele lapha we-SARS-CoV-2 coronavirus inactivation eyenziwe nguWelch et al. (2020) ngezansi:

Amaseli Namagciwane

Amaseli e-Vero E6 (i-Vero C1008; i-ATCC CRL-1586) akhuliswa kumandla aphansi abalulekile we-Eagle (MEM) afakwa nge-10% (v / v) fetal calf serum (FCS). Igciwane elisetshenzisiwe kwakuyinkinga ye-SARS-CoV-2 hCOV-19 / England / 2/2020, ihlukaniswe yi-PHE eqenjini lokuqala lesiguli e-UK ngomhlaka 29/01/2020. Leli gciwane litholakale endimeni 1 futhi lasetshenziselwa ukwenza izifundo zingasebenzi endimeni 2 noma 3.
Ngama-reagents namakhemikhali asetshenziselwa ukusebenza kwe-SARS-CoV-2 kanye nokususwa kwe-cytotoxicity esebenzayo sicela ubheke umbiko wesayensi kaWelch et al. (2020).

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

Ukungasebenzi kwegciwane ngama-detergents – I-SARS-CoV-2 Coronavirus Inactivation protocol eyenziwe nguWelch et al. 2020

Ukusebenza kwe-SARS-CoV-2

Okwemikhiqizo yezentengiselwano, ukulungiselela amagciwane (uketshezi lwesiko lezicubu, izihloko ezisukela ku-1 × 106 kuya ku-1 × 108 I-PFU / ml) iphathwe ngokuphindwe kathathu ngama-reagents ekugxileni nasezikhathini zokuxhumana ezinconywe kumiyalo yabakhiqizi yokusetshenziswa, lapho kutholakala khona, noma ukugxila kanye nezikhathi ezicelwe ngokukhethekile ngokuhlolwa kwamalabhorethri. Lapho ububanzi bokugxila bunikezwe ngumenzi, isilinganiso esiphansi somkhiqizo kwisampula sahlolwa (isb. Ukuhlushwa okunconywe okuphansi kakhulu komkhiqizo wokuhlola). Ama-reagents amashubhu ezokuthutha ahlolwa kusetshenziswa isilinganiso sevolumu eyodwa yoketshezi lwesiko lezicubu kuya kumavolumu ayishumi we-reagent, ngaphandle kokuthi isilinganiso sevolumu yesampula uketshezi kuya kuma-reagent sicacisiwe ngumenzi. Ama-detergent, ama-fixatives nama-solvents ahlolwe ezindaweni ezibonisiwe zezikhathi ezikhonjisiwe. Zonke izinyathelo zokungasebenzi zenziwa emazingeni okushisa egumbi (18 - 25 ° C). Ukuhlolwa kwezinye izinhlobo zesampula, igciwane lafakwa ku-matrix yesampula ekhonjisiwe ngesilinganiso esingu-1: 9, bese liphathwa ngama-reagents wokuhlola njengangenhla. Zonke izivivinyo zazibandakanya amasampula wokulawulwa okuphindwe kabili okulawulwa okuphindwe kabili ngevolumu elinganayo ye-PBS esikhundleni se-reagent test. Ngokushesha okulandela isikhathi sokuxhumana esidingekayo, i-1mL yesampula ephethwe yacutshungulwa kusetshenziswa i-matrix yokuhlunga ekhethwe ngokufanele. Ukususwa kwe-reagent kokuhlolwa kokungasebenzi kwenziwa ngefomethi yekholomu enkulu ye-spin kusetshenziswa i-Pierce 4mL Detergent Removal Spin Columns (Thermo Fisher), noma ngokugcwalisa amakholomu we-Pierce 10mL umthamo centrifuge angenalutho (Thermo Fisher) nge-SM2 Bio-Beads, Sephacryl S-400HR noma I-Sephadex LH-20 ukunikeza ubuhlalu / i-resin egcwele i-4mL. Ukuze kuhlanzwe kusetshenziswa izihlungi ze-Amicon, amasampula ama-2 × 500μl ahlanzwa kusetshenziswa izihlungi ezimbili ze-centrifugal ngendlela echazwe ngaphambili, bese ahlanganiswa ndawonye. Okwe-formaldehyde ne-formaldehyde okususwa kwe-glutaraldehyde, isihlungi esisodwa sasetshenziswa ngevolumu yesampula engu-1 × 500μl, savuselelwa ngemuva kokucutshungulwa ku-500μl PBS, futhi sangezwa ku-400ul MEM / 5% FBS. Yokwenza kungasebenzi kwegciwane VialTweeter at the ultrasonic processor UP200STama-monolayers, 12.5 cm2 amabhodlela wamaseli we-Vero E6 (2.5 × 106 amaseli / i-flask ku-2.5mL MEM / 5% FBS) atheleleke ku-MOI 0.001 futhi afakwa ku-37 ° C / 5% CO2 amahora angama-24. I-supernatant isusiwe, futhi amaseli alungiswa kusetshenziswa i-5mL ye-formaldehyde, noma i-formaldehyde ne-glutaraldehyde ekamelweni lokushisa imizuzu eyi-15 noma engu-60. I-fixative isusiwe, futhi ama-monolayers ageza amahlandla amathathu nge-PBS ngaphambi kokukhuhla amaseli abe yi-1mL MEM / 5% FBS ne-sonicated (3 × 10 second on, 10 seconds off at 100% power and amplitude) using UP200St with VialTweeter attachment (Hielscher Ultrasound Ubuchwepheshe). Ama-supernatants acacisiwe ngokufakwa kwe-centrifuging ku-3000 × g imizuzu eyi-10.

I-VialTweeter ye-Sonication Enamandla yamaViva avaliwe

I-VialTweeter ye-Sonication Enamandla Yezitsha Ezivaliwe

Isicelo solwazi




Qaphela kwethu Inqubomgomo yobumfihlo.


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

Imininingwane ye-reagent esetshenziselwe i-SARS-CoV-2 coronavirus inactivation ne-ultrasonic VialTweeter ngokuya ngephrothokholi kaWelch et al. 2020

I-protocol ephelele efaka ukusetshenziswa kwe-Hielscher VialTweeter ingatholakala lapha:
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!)

I-VialTweeter ye-ultrasonic virus inactivation

Izinzuzo ze-VialTweeter shazi

  • I-Sonication ye-10 izitsha ngesikhathi esisodwa
  • Akukho ukungcoliswa okuphambene
  • Akukho ukulahleka kwesampula
  • Ukuqoshwa kwedatha okuzenzakalelayo
  • Kulula Futhi Kuphephile Ukusebenza
IVialTweeter nayo isetshenziselwa i-

  • I-Cell Lysis
  • Ukuphazamiseka kwezinhlayiyana zegciwane
  • Ukukhishwa kwe-Nucleic acid: Ukuhlukaniswa kwe-DNA / RNA
  • Ukuhlukaniswa kwe-DNA / RNA
  • Ukuxazululwa kweLysate
  • Ama-Dismembrator we-Ultrasonic aphezulu kanye nokuphazamiseka kwamaseli

    I-multi-sample ultrasonicator VialTweeter ingesinye sezixazululo eziningi ze-ultrasonic zokulungiselela isampula kumalabhorethri wezinto eziphilayo, amakhemikhali namakhemikhali. I-Hielscher Ultrasonics inikeza i-ultrasonic dismembrator ekahle yohlelo lwakho lokusebenza, isb. I-cell lysis, i-cell extraction, i-homogenization yezicubu, i-lysate solubilization, ukuncibilika, isampula yokunciphisa njll.
    Sazise ukuthi mangaki amasampula okufanele usebenze ngehora nosuku, uma ukhetha i-sonication eqondile noma engaqondile nokuthi ilitshe le-ultrasonic sample treatment liyini. Sizokuncoma iyunithi ye-ultrasonic efaneleke kakhulu emsebenzini wakho wansuku zonke!
    Amaprosesa we-Hielscher Ultrasonics 'digital ultrasonic afakwe isoftware ehlakaniphile, ukuqoshwa kwedatha okuzenzakalelayo, izinketho ezilula zangaphambi kokusetha zokulawula izinga lokushisa, ubude be-sonication, imodi yomjikelezo / yokushaya kwenhliziyo kanye nokukhanyisa kwesampula nokulawula okukude kwesiphequluli. Silwela ukwenza amadivayisi wethu we-ultrasonic abe smart ngangokunokwenzeka ukuze ucwaningo lwakho nomsebenzi wakho ube lula futhi uphumelele ngangokunokwenzeka.
    Chofoza lapha, ukuthola ezinye izinhlelo zokusebenza kufaka phakathi izivumelwano zokulungiselela isampula ye-ultrasonic nge-VialTweeter!

    Xhumana nathi! / Cela Us!

    Cela ulwazi oluthe xaxa

    Uyacelwa ukuthi usebenzise ifomu elingezansi ukucela ulwazi olungeziwe mayelana nezinhlelo zokusebenza ze-ultrasonic, izinhlelo zokusebenza nentengo. Sizokujabulela ukuxoxa ngenqubo yakho nawe futhi sikunikeze uhlelo lwe-ultrasonic oluhlangabezana nezidingo zakho!









    Sicela uqaphele wethu Inqubomgomo yobumfihlo.


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

    I-Hielscher Ultrasonics ikhiqiza ama-homogenizers asebenza ngokusebenza okuphezulu wokuxuba izinhlelo, ukusabalalisa, ukumisa nokukhipha ilebhu, umshayeli kanye nesikali sezimboni.

    Izincwadi / Izikhombo



    Amaqiniso Okufanele Ukwazi

    Ayini amaseli eVero?

    I-Vero E6, eyaziwa nangokuthi i-Vero C1008 (i-ATCC No. CRL-1586) ilayini weseli eliqhamuka ku-Vero 76 futhi isetshenziswa ocwaningweni lwe-SARS-CoV ne-SARS-CoV-2 coronaviruses. Amaseli e-Vero ayinzalo yamaseli asetshenziswa kumasiko amaseli. I-'Vero’ Uhlu lozalo lwaluhlukaniswe namaseli ezinso epithelial akhishwe enkawini eluhlaza yase-Afrika (Chlorocebus sp.).
    Amaseli e-Vero E6 akhombisa ukuvinjelwa kokuxhumana okuthile, ngakho-ke kufanelekile ukusabalalisa amagciwane aziphindaphinda kancane. Imigqa yamaseli e-Vero E6 ijwayele ukusetshenziselwa ukuphenya i-cytopathology yama-coronaviruses SARS-CoV kanye ne-SARS-CoV-2 njengoba amaseli e-Vero (amaseli wezinso zezinkawu zase-Afrika) ekhombisa ukuvezwa okuningi kwe-angiotensin-converting enzyme 2 (ACE2) receptors. Ama-receptors e-ACE2 ayisiza esikhulu semikhumbi se-SARS-CoV-2 coronavirus.
    Isibonelo, u-Ogando et al. (2020) bathole ukuthi i-SARS-CoV-2 – uma kuqhathaniswa ne-SARS-CoV – kukhiqize amazinga aphakeme we-intracellular virus RNA, kepha ngokumangazayo cishe cishe ama-50 inzalo yegciwane lesandulela ngculazi engatheleleki yatholwa endaweni yesiko. Ngaphezu kwalokho, banqume ukuthi ukuzwela kwamagciwane amabili kuma-inhibitor amathathu asungulwe we-coronavirus replication (i-Remdesivir, i-Alisporivir ne-chloroquine) kuyafana kakhulu, kepha ukuthi ukutheleleka kwe-SARS-CoV-2 bekuzwela kakhulu ekwelashweni kwangaphambili kwamangqamuzana ane-interferon ene-pegylated i-alpha. Umehluko obalulekile phakathi kwamagciwane amabili iqiniso lokuthi – ngokudlula kumaseli we-Vero E6 – I-SARS-CoV-2 ngokusobala ingaphansi kwengcindezi yokukhetha enamandla yokuthola ukuguqulwa okuguqukayo kuhlobo lwayo lwe-spike protein. Lokhu kuguquka kuguqula noma kususe indawo yokuhlanza 'efana ne-furin-like cleavage' esifundeni esixhuma izizinda ze-S1 ne-S2 futhi kuholele ekushintsheni okuvelele kakhulu kwe-phenotypic ekuhloleni kwe-plaque.