1. Mayelana nokukhiqizwa kwamapayipi okuthatha amasampula egciwane
Amashubhu okuthatha amasampula egciwane angowemikhiqizo yedivayisi yezokwelapha. Iningi labakhiqizi basekhaya libhaliswe ngokwemikhiqizo yekilasi lokuqala, kanti zimbalwa izinkampani ezibhaliswe ngokwemikhiqizo yekilasi lesibili. Muva nje, ukuze kuhlangatshezwane nezidingo eziphuthumayo zaseWuhan nakwezinye izindawo, izinkampani eziningi zithathe "isiteshi esiphuthumayo" "Faka isicelo semvume yokurekhoda yekilasi lokuqala. Ishubhu lokuthatha amasampula egciwane lakhiwe nge-swab yokuthatha amasampula, ikhambi lokulondoloza igciwane kanye nokupakishwa kwangaphandle. Njengoba kungekho zindinganiso zikazwelonke ezihlanganisiwe noma indinganiso yemboni, imikhiqizo yabakhiqizi abahlukahlukene iyahlukahluka kakhulu.
1. I-swab yokuthatha amasampula: I-swab yokuthatha amasampula ithinta ngqo indawo yokuthatha amasampula, futhi izinto zekhanda lokuthatha amasampula zihlobene kakhulu nokutholwa okulandelayo. Ikhanda le-swab yokuthatha amasampula kufanele lenziwe nge-fiber yokwenziwa ye-Polyester (PE) noma i-Rayon (fiber eyenziwe ngumuntu). Iziponji ze-calcium alginate noma izinduku zokhuni (kufaka phakathi izinduku ze-bamboo) azikwazi ukusetshenziswa, futhi izinto zekhanda le-swab azikwazi ukuba imikhiqizo yekotoni. Ngenxa yokuthi i-fiber yekotoni inomumo oqinile weprotheni, akulula ukungena esixazululweni sokugcina esilandelayo; futhi lapho induku yokhuni noma induku ye-bamboo equkethe i-calcium alginate kanye nezingxenye zokhuni iphukile, ukucwiliswa esixazululweni sokugcina nakho kuzomunca iphrotheni, futhi kungavimbela ukusabela okulandelayo kwe-PCR. Kunconywa ukusebenzisa imicu yokwenziwa njenge-fiber ye-PE, i-fiber ye-polyester kanye ne-fiber ye-polypropylene ezintweni zekhanda le-swab. Imicu yemvelo efana nekotini ayinconywa. Imicu ye-nylon nayo ayinconywa ngoba imicu ye-nylon (efana namakhanda ebhulashi lamazinyo) imunca amanzi. Ayilungile, okuholela ekwandeni okunganele kwevolumu yokuthatha amasampula, okuthinta izinga lokutholwa. Isiponji se-calcium alginate asivunyelwe ezintweni zokuthatha amasampula! Isibambo se-swab sinezinhlobo ezimbili: esiphukile nesakhelwe ngaphakathi. I-swab ephukile ifakwa epayipini lokugcina ngemva kokuthatha isampula, futhi isivalo setyhubhu siphukile ngemva kokwephuka sisuka endaweni eseduze nekhanda lokuthatha isampula; i-swab eyakhelwe ngaphakathi ifaka ngqo i-swab yokuthatha isampula epayipini lokugcina ngemva kokuthatha isampula, futhi isembozo setyhubhu lokugcina sakhiwe ngaphakathi. Qondanisa imbobo encane nengxenye ephezulu yesibambo bese uqinisa isembozo setyhubhu. Uma kuqhathaniswa lezi zindlela ezimbili, lokhu kokugcina kuphephile. Uma i-swab ephukile isetshenziswa kanye ne-tube yokugcina encane, ingabangela ukuchitheka koketshezi epayipini lapho iphukile, futhi kufanele kunakwe ngokugcwele ingozi yokungcola okubangelwa ukusetshenziswa okungafanele komkhiqizo. Kunconywa ukusebenzisa ithubhu elingaphandle le-polystyrene (PS) elingenalutho noma ithubhu eliqhekeka nge-polypropylene (PP) lezinto ze-swab handle. Kungakhathaliseki ukuthi kusetshenziswa yiphi impahla, izithasiselo ze-calcium alginate azikwazi ukungezwa; izinti zokhuni noma izinti ze-bamboo. Ngamafuphi, i-swab yokuthatha isampula kufanele iqinisekise inani lesampula kanye nenani lokukhululwa, futhi izinto ezikhethiwe akumele zibe nezinto ezithinta ukuhlolwa okulandelayo.
2. Isixazululo sokulondolozwa kwegciwane: Kunezinhlobo ezimbili zezixazululo zokulondolozwa kwegciwane ezisetshenziswa kabanzi emakethe, esinye siyisisombululo sokulondolozwa kwegciwane esishintshiwe ngokusekelwe endaweni yokuthutha, kanti esinye siyisisombululo esishintshiwe se-nucleic acid extraction lysate.
Ingxenye eyinhloko yalokhu kokuqala i-Eagle's basic culture medium (MEM) noma usawoti olinganiselayo kaHank, ofakwa nosawoti, ama-amino acid, amavithamini, i-glucose kanye namaprotheni adingekayo ukuze igciwane lisinde. Lesi sixazululo sokugcina sisebenzisa usawoti we-phenol red sodium njengesibonakaliso kanye nesisombululo. Uma inani le-pH lingu-6.6-8.0, isisombululo siba pinki. I-glucose edingekayo, i-L-glutamine kanye namaprotheni kuyangezwa esixazululweni sokugcina. Iphrotheni inikezwa ngesimo se-fetal bovine serum noma i-bovine serum albumin, engazinzisa igobolondo lephrotheni legciwane. Ngenxa yokuthi isisombululo sokugcina sicebile ngezakhamzimba, siyasiza ekusindeni kwegciwane kodwa futhi siwusizo ekukhuleni kwamagciwane. Uma isisombululo sokugcina singcoliswe amagciwane, sizophindaphindeka ngobuningi. I-carbon dioxide kuma-metabolites ayo izobangela ukuthi isisombululo sokugcina i-pH iwele kusuka kupinki Iphenduka ibe phuzi. Ngakho-ke, abakhiqizi abaningi bangeze izithako zokulwa namagciwane ekwakhekeni kwabo. Ama-antibacterial agents anconyiwe yi-penicillin, i-streptomycin, i-gentamicin kanye ne-polymyxin B. I-Sodium azide kanye ne-2-methyl akunconywa. Ama-inhibitors afana ne-4-methyl-4-isothiazolin-3-one (MCI) kanye ne-5-chloro-2-methyl-4-isothiazolin-3-one (CMCI) ngoba lezi zingxenye zinomthelela ekusabeleni kwe-PCR. Njengoba isampula enikezwe yilesi sixazululo sokulondoloza ngokuyisisekelo iyigciwane eliphilayo, ubuqambi besampula bungagcinwa ngezinga elikhulu kakhulu, futhi ingasetshenziswa hhayi kuphela ekukhishweni nasekutholweni kwama-nucleic acid egciwane, kodwa futhi nasekutshalweni nasekuhlukanisweni kwama-virus. Kodwa-ke, kufanele kuqashelwe ukuthi uma isetshenziselwa ukuthola, ukukhishwa nokuhlanzwa kwe-nucleic acid kumele kwenziwe ngemva kokungasebenzi.
Olunye uhlobo lwesisombululo sokulondolozwa esilungiselelwe ngokusekelwe ku-lysate yokukhipha i-nucleic acid, izingxenye eziyinhloko usawoti olinganiselayo, i-EDTA chelating agent, usawoti we-guanidine (njenge-guanidine isothiocyanate, i-guanidine hydrochloride, njll.), i-anionic surfactant (njenge-dodecane Sodium sulfate), ama-surfactants e-cationic (njenge-tetradecyltrimethylammonium oxalate), i-phenol, i-8-hydroxyquinoline, i-dithiothreitol (DTT), i-proteinase K nezinye izingxenye. Lesi sixazululo sokugcina siwukuqhekeza ngqo igciwane ukuze kukhishwe i-nucleic acid futhi kuqedwe i-RNase. Uma sisetshenziswa kuphela ku-RT-PCR, sifaneleka kakhulu, kodwa i-lysate ingalivimba igciwane. Lolu hlobo lwesampula alukwazi ukusetshenziselwa ukuhlukaniswa kwamasiko egciwane.
I-ejenti ye-metal ion chelating esetshenziswa esixazululweni sokulondolozwa kwegciwane inconywa ukuthi isebenzise usawoti we-EDTA (njenge-dipotassium ethylenediaminetetraacetic acid, i-disodium ethylenediaminetetraacetic acid, njll.), futhi akukhuthazwa ukusebenzisa i-heparin (njenge-sodium heparin, i-lithium heparin), ukuze kungathinti ukutholwa kwe-PCR.
3. Ishubhu lokugcina: Izinto zeshubhu lokugcina kufanele zikhethwe ngokucophelela. Kukhona idatha ephakamisa ukuthi i-polypropylene (i-Polypropylene) ihlobene nokumuncwa kwe-nucleic acid, ikakhulukazi lapho kugxilwe khona ama-ion aphezulu, i-polyethylene (i-Polyethylene) ithandwa kakhulu kune-polypropylene (i-Polypropylene). Kulula ukubamba i-DNA/RNA. Ipulasitiki ye-polyethylene-propylene polymer (i-Polyallomer) kanye nezinye izitsha zepulasitiki ze-polypropylene (i-Polypropylene) ezicutshungulwe ngokukhethekile zifaneleka kakhulu ukugcina i-DNA/RNA. Ngaphezu kwalokho, uma usebenzisa ishubhu eliphukayo, ishubhu lokugcina kufanele lizame ukukhetha isitsha esinokuphakama okungaphezu kuka-8 cm ukuvimbela okuqukethwe ukuthi kungachitheki futhi kungcoliswe lapho ishubhu liphukile.
4. Isixazululo sokugcina amanzi: Amanzi ahlanzekile kakhulu asetshenziselwa isixazululo sokugcina amanzi kufanele ahlungwe nge-membrane ye-ultrafiltration enesisindo sama-molecule esingu-13,000 ukuqinisekisa ukususwa kokungcola kwe-polymer emithonjeni yezinto eziphilayo, njenge-RNase, i-DNase, ne-endotoxin, futhi ukuhlanzwa okuvamile akukhuthazwa. Amanzi noma amanzi acwengekile.
2. Ukusetshenziswa kwamapayipi okuthatha amasampula egciwane
Ukusampula kusetshenziswa ithubhu lokuhlola igciwane kuhlukaniswe kakhulu ukusampula kwe-oropharyngeal kanye nokusampula kwe-nasopharyngeal:
1. Ukuhlolwa kwe-Oropharyngeal: Okokuqala cindezela ulimi ngomshini wokucindezela ulimi, bese welula ikhanda le-swab yokuhlolwa emphinjeni ukuze usule amathonsil omphimbo asemaceleni omabili kanye nodonga lwe-pharyngeal olungemuva, bese usula udonga lwe-pharyngeal olungemuva ngamandla amancane, gwema ukuthinta iyunithi yolimi.
2. Ukuthatha isampula ye-nasopharyngeal: kala ibanga ukusuka esiqongweni sekhala kuya esiqongweni sendlebe nge-swab bese uphawula ngomunwe, faka i-swab yesampula emgodini wekhala ubheke ekhaleni eliqondile (ubuso), i-swab kufanele yelule okungenani ingxenye yobude be-lobe yendlebe iye esiqongweni sekhala, Shiya i-swab ekhaleni imizuzwana eyi-15-30, ujikeleze kancane izikhathi ezi-3-5, bese ukhipha i-swab.
Akunzima ukubona ngendlela yokusebenzisa, kungakhathaliseki ukuthi kuyi-oropharyngeal swab noma i-nasopharyngeal swab, ukusampula kuwumsebenzi wobuchwepheshe, onzima futhi ongcolile. Ikhwalithi yesampula eqoqwe ihlobene ngqo nokutholwa okulandelayo. Uma isampula eqoqwe inomthamo wegciwane. Kuphansi, kulula ukubangela imiphumela emibi engamanga, kunzima ukuqinisekisa ukuxilongwa.
Isikhathi sokuthunyelwe: Juni-21-2020
