1. Malunga nokuveliswa kweetyhubhu zokuthatha iisampulu zeentsholongwane
Iityhubhu zokuthatha iisampulu zeentsholongwane zezemveliso zezixhobo zonyango. Uninzi lwabavelisi basekhaya babhaliswe ngokweemveliso zodidi lokuqala, kwaye zimbalwa iinkampani ezibhaliswe ngokweemveliso zodidi lwesibini. Kutshanje, ukuze kuhlangatyezwane neemfuno zongxamiseko zaseWuhan nakwezinye iindawo, iinkampani ezininzi zithathe "itshaneli yongxamiseko" "Faka isicelo semvume yokurekhoda yodidi lokuqala. Ityhubhu yokuthatha iisampulu zeentsholongwane yenziwe ngesampulu yesampulu, isisombululo sokugcina iintsholongwane kunye nokupakishwa kwangaphandle. Ekubeni kungekho mgangatho kazwelonke udibeneyo okanye umgangatho weshishini, iimveliso zabavelisi abahlukeneyo zahluka kakhulu.
1. I-swab yokuvavanya: I-swab yokuvavanya idibana ngqo nendawo yokuvavanya, kwaye izinto zentloko yokuvavanya zinxulumene kakhulu nokufunyanwa okulandelayo. Intloko ye-swab yokuvavanya kufuneka yenziwe nge-fiber yokwenziwa ye-Polyester (PE) okanye i-Rayon (i-fiber eyenziwe ngumntu). I-sponge ye-calcium alginate okanye ii-swab ze-wooden stick (kubandakanya ii-bamboo sticks) azinakusetyenziswa, kwaye izinto zentloko ye-swab azinakuba ziimveliso zekotoni. Ngenxa yokuba i-cotton fiber ine-adsorption enamandla yeproteni, akulula ukuyifaka kwisisombululo sokugcina esilandelayo; kwaye xa intonga yomthi okanye intonga ye-bamboo equlethe i-calcium alginate kunye nezinto zomthi iphukile, ukuntywila kwisisombululo sokugcina kuya kutsala iproteni, kwaye kuya kuyithintela impendulo elandelayo ye-PCR. Kucetyiswa ukusebenzisa ii-synthetic fibers ezifana ne-PE fiber, i-polyester fiber kunye ne-polypropylene fiber kwizinto zentloko ye-swab. Ii-fiber zendalo ezifana ne-cotton azikhuthazwa. Ii-fiber ze-nylon nazo azikhuthazwa kuba ii-fiber ze-nylon (ezifana neentloko zamazinyo) zifunxa amanzi. Zimbi, nto leyo ebangela ukuba umthamo we-sampling unganeli, nto leyo echaphazela izinga lokufunyanwa. I-calcium alginate sponge ayivumelekanga kwizinto ze-sampling swab! Isiphatho se-swab sineentlobo ezimbini: esaphukileyo nesakhelwe ngaphakathi. I-swab ephukileyo ifakwa kwityhubhu yokugcina emva kokuthatha isampuli, kwaye isivalo setyhubhu siyaphuka emva kokuba siphuke kwindawo ekufutshane nentloko yesampuli; i-swab eyakhelwe ngaphakathi ifaka ngqo i-swab yesampuli kwityhubhu yokugcina emva kokuthatha isampuli, kwaye isigqubuthelo setyhubhu yokugcina sakhiwe ngaphakathi. Lungelelanisa umngxuma omncinci nomphezulu wesiphatho kwaye uqinise isigqubuthelo setyhubhu. Xa kuthelekiswa ezi ndlela zimbini, le yokugqibela ikhuselekile. Xa i-swab ephukileyo isetyenziswa kunye netyhubhu yokugcina encinci, inokubangela ukutshiza kolwelo kwityhubhu xa iphuke, kwaye kufuneka kunikwe ingqalelo epheleleyo kumngcipheko wokungcola okubangelwa kukusetyenziswa okungafanelekanga kwemveliso. Kucetyiswa ukusebenzisa ityhubhu engenanto ye-polystyrene (PS) ekhutshiweyo okanye ityhubhu yokukrala injection ye-polypropylene (PP) kwizinto zesiphatho se-swab. Nokuba kusetyenziswa ziphi na izinto, izongezo ze-calcium alginate azinakongezwa; iintonga zomthi okanye iintonga ze-bamboo. Ngamafutshane, i-swab yesampuli kufuneka iqinisekise ubungakanani besampulu kunye nobungakanani bokukhululwa, kwaye izixhobo ezikhethiweyo akufuneki zibe nezinto ezichaphazela uvavanyo olulandelayo.
2. Isisombululo sokugcina iintsholongwane: Kukho iintlobo ezimbini zezisombululo zokugcina iintsholongwane ezisetyenziswa kakhulu kwimarike, esinye sisisombululo sokugcina iintsholongwane esiguqulweyo ngokusekelwe kwindlela yokuthutha, kwaye esinye sisisombululo esiguqulweyo se-nucleic acid extraction lysate.
Eyona nto iphambili kwi-Eagle's basic culture medium (MEM) okanye ityuwa elinganisiweyo yeHank, eyongezwa kunye neetyuwa, ii-amino acids, iivithamini, iglucose kunye neeproteni ezifunekayo ukuze intsholongwane iphile. Esi sisombululo sokugcina sisebenzisa ityuwa yesodium ebomvu ye-phenol njengesalathisi kunye nesisombululo. Xa ixabiso le-pH liyi-6.6-8.0, isisombululo siba pinki. Iglucose efunekayo, i-L-glutamine kunye neproteni zongezwa kwisisombululo sokugcina. Iproteni inikezelwa ngohlobo lwe-fetal bovine serum okanye i-bovine serum albumin, enokuzinzisa igobolondo leproteni yentsholongwane. Ngenxa yokuba isisombululo sokugcina sityebile kwizakhamzimba, siluncedo ekusindeni kwentsholongwane kodwa sikwaluncedo ekukhuleni kweebhaktheriya. Ukuba isisombululo sokugcina singcoliswe ziibhaktheriya, siya kuphinda-phinda ngobuninzi. I-carbon dioxide kwi-metabolites zayo iya kubangela ukuba isisombululo sokugcina i-pH iwele kwi-pink Ijika ibe tyheli. Ke ngoko, uninzi lwabavelisi bongeze izithako zokulwa ne-antibacterial kwifomyula yabo. Iiarhente ezicetyiswayo zokulwa nentsholongwane yipenicillin, streptomycin, gentamicin kunye nepolymyxin B. I-Sodium azide kunye ne-2-methyl azikhuthazwa. Izithinteli ezifana ne-4-methyl-4-isothiazolin-3-one (MCI) kunye ne-5-chloro-2-methyl-4-isothiazolin-3-one (CMCI) kuba ezi zinto zinefuthe kwindlela esebenza ngayo i-PCR. Ekubeni isampuli ebonelelwe sesi sisombululo sokulondoloza ngokusisiseko yintsholongwane ephilayo, ubunyani besampuli bunokugcinwa kakhulu, kwaye bungasetyenziswa kungekuphela nje ekukhupheni nasekufumaneni ii-nucleic acids zentsholongwane, kodwa nasekukhuliseni nasekuhlukaniseni iintsholongwane. Nangona kunjalo, kufuneka kuqatshelwe ukuba xa isetyenziselwa ukufumanisa, ukukhupha kunye nokucoca i-nucleic acid kufuneka kwenziwe emva kokungasebenzi.
Olunye uhlobo lwesisombululo sokugcina esilungiselelwe ngokusekelwe kwi-lysate yokukhupha i-nucleic acid, izinto eziphambili ziityuwa ezilinganiselayo, i-EDTA chelating agent, ityuwa ye-guanidine (efana ne-guanidine isothiocyanate, i-guanidine hydrochloride, njl.njl.), i-anionic surfactant (efana ne-dodecane Sodium sulfate), ii-cationic surfactants (efana ne-tetradecyltrimethylammonium oxalate), i-phenol, i-8-hydroxyquinoline, i-dithiothreitol (DTT), i-proteinase K kunye nezinye izinto. Esi sisombululo sokugcina sisokunqumla ngqo intsholongwane ukuze ikhuphe i-nucleic acid kwaye isuse i-RNase. Ukuba isetyenziselwa i-RT-PCR kuphela, ifanelekile ngakumbi, kodwa i-lysate inokuyicima intsholongwane. Olu hlobo lwesampulu alunakusetyenziselwa ukwahlula inkcubeko yentsholongwane.
I-arhente ye-metal ion chelating esetyenziswa kwisisombululo sokugcina iintsholongwane iyacetyiswa ukuba isebenzise iityuwa ze-EDTA (ezifana ne-dipotassium ethylenediaminetetraacetic acid, i-disodium ethylenediaminetetraacetic acid, njl.njl.), kwaye akukhuthazwa ukuba kusetyenziswe i-heparin (ezifana ne-sodium heparin, i-lithium heparin), ukuze ingachaphazeli ukufunyanwa kwe-PCR.
3. Ityhubhu yokugcina: Izinto ezikwityhubhu yokugcina kufuneka zikhethwe ngononophelo. Kukho idatha ebonisa ukuba i-polypropylene (i-Polypropylene) inxulumene nokufunxwa kwe-nucleic acid, ingakumbi xa i-ion concentration iphezulu, i-polyethylene (i-Polyethylene) ikhethwa kakhulu kune-polypropylene (i-Polypropylene). Kulula ukuyiqonda i-DNA/RNA. Iplastiki ye-polyethylene-propylene polymer (i-Polyallomer) kunye nezinye izikhongozeli zeplastiki ze-polypropylene (i-Polypropylene) ezicutshungulwe ngokukodwa zifanelekile ngakumbi ukugcina i-DNA/RNA. Ukongeza, xa usebenzisa i-swab enokuqhekeka, ityhubhu yokugcina kufuneka izame ukukhetha isitya esinobude obungaphezulu kwe-8 cm ukuthintela ukuba umxholo ungatshizwa kwaye ungcoliswe xa i-swab iphukile.
4. Isisombululo sokugcina amanzi: Amanzi acocekileyo kakhulu asetyenziselwa isisombululo sokugcina imveliso kufuneka ahluzwe nge-membrane ye-ultrafiltration enobunzima be-molecular be-13,000 ukuqinisekisa ukususwa kokungcola kwe-polymer kwimithombo yebhayoloji, efana ne-RNase, i-DNase, kunye ne-endotoxin, kwaye ukucocwa okuqhelekileyo akukhuthazwa. Amanzi okanye amanzi acocekileyo.
2. Ukusetyenziswa kweetyhubhu zokuthatha iisampulu zeentsholongwane
Ukusampula kusetyenziswa ityhubhu yokuvavanya intsholongwane kwahlulwe ngokubanzi kwi-oropharyngeal sampling kunye ne-nasopharyngeal sampling:
1. Ukuthathwa kwesampulu ye-Oropharyngeal: Okokuqala cinezela ulwimi ngesithinteli solwimi, uze wolule intloko yesampulu yesampulu emqaleni ukuze usule iitoni ze-pharyngeal ezikumacala omabini kunye nodonga lwe-pharyngeal olungasemva, uze usule udonga lwe-pharyngeal olungasemva ngamandla alula, uphephe ukuchukumisa iyunithi yolwimi.
2. Ukuthatha isampuli ye-nasopharyngeal: linganisa umgama ukusuka kwincam yempumlo ukuya kwi-lobe yendlebe nge-swab uze uphawule ngomnwe, faka i-sampling swab kwi-nose cavity kwicala lempumlo ethe nkqo (ubuso), i-swab kufuneka yolulele ubuncinane isiqingatha sobude be-lobe yendlebe ukuya kwincam yempumlo. Shiya i-swab empumlweni imizuzwana eli-15-30, ujikeleze kancinci amaxesha ama-3-5, uze urhoxe i-swab.
Akunzima ukubona kwindlela yokusebenzisa, nokuba yi-oropharyngeal swab okanye i-nasopharyngeal swab, ukuthathwa kwesampulu ngumsebenzi wobugcisa, onzima kwaye ungcolisiwe. Umgangatho wesampulu eqokelelweyo unxulumene ngokuthe ngqo nokufunyanwa okulandelayo. Ukuba isampuli eqokelelweyo inomthwalo wentsholongwane. Iphantsi, kulula ukubangela iziphumo ezingezizo, kunzima ukuqinisekisa ukuxilongwa.
Ixesha lokuthumela: Juni-21-2020
