1. E pili ana i ka hana ʻana o nā ʻōmole laʻana virus
No nā huahana mea lapaʻau nā paipu laʻana virus. Ua hoʻopaʻa inoa ʻia ka hapa nui o nā mea hana kūloko e like me nā huahana papa mua, a he kakaikahi nā ʻoihana i hoʻopaʻa inoa ʻia e like me nā huahana papa lua. I kēia manawa, i mea e hoʻokō ai i nā pono pilikia o Wuhan a me nā wahi ʻē aʻe, ua lawe nā ʻoihana he nui i ke "kahawai pilikia" "E noi no ka ʻae palapala papa mua." Ua hana ʻia ka paipu laʻana virus me kahi swab laʻana, kahi hopena mālama virus a me ka ʻōpala waho. No ka mea ʻaʻohe kūlana aupuni i hoʻokahi a i ʻole ke kūlana ʻoihana, ʻokoʻa loa nā huahana o nā mea hana like ʻole.
1. ʻŌpala laʻana: Hoʻopili pololei ka ʻōpala laʻana i kahi laʻana, a pili pono ka mea o ke poʻo laʻana i ka ʻike ʻana ma hope. Pono ke poʻo ʻōpala laʻana e hana ʻia me ka fiber synthetic Polyester (PE) a i ʻole Rayon (fiber hana kanaka). ʻAʻole hiki ke hoʻohana ʻia ka huʻahuʻa calcium alginate a i ʻole nā ʻōpala lāʻau (me nā lāʻau ʻohe), a ʻaʻole hiki ke lilo i nā huahana pulupulu ka mea o ke poʻo ʻōpala. No ka mea he adsorption ikaika ka fiber pulupulu i ka protein, ʻaʻole maʻalahi ke elute i loko o ka hopena mālama ma hope; a i ka wā e haki ai kahi lāʻau lāʻau a i ʻole ka lāʻau ʻohe e loaʻa ana ka calcium alginate a me nā ʻāpana lāʻau, ʻo ka hoʻomoʻa ʻana i loko o ka hopena mālama e adsorb pū i ka protein, a hiki iā ia ke kāohi i ka hopena PCR ma hope. Manaʻo ʻia e hoʻohana i nā fiber synthetic e like me ka fiber PE, polyester fiber a me ka polypropylene fiber no ka mea o ke poʻo ʻōpala. ʻAʻole ʻōlelo ʻia nā fibers kūlohelohe e like me ka pulupulu. ʻAʻole ʻōlelo ʻia nā fibers Nylon no ka mea ʻo nā fibers nylon (like me nā poʻo niho palaki) e omo i ka wai. ʻAʻole maikaʻi, e hopena ana i ka lawa ʻole o ka nui o ka laʻana, e hoʻopilikia ana i ka helu ʻike. Ua pāpā ʻia ka huʻahuʻa calcium alginate no ka laʻana ʻana i ka mea swab! ʻElua ʻano o ka lima swab: haki a kūkulu ʻia. Hoʻokomo ʻia ka swab haki i loko o ka paipu mālama ma hope o ka laʻana ʻana, a ua haki ke poʻi paipu ma hope o ka haki ʻana mai ke kūlana kokoke i ke poʻo laʻana; hoʻokomo pololei ka swab i kūkulu ʻia i ka swab laʻana i loko o ka paipu mālama ma hope o ka laʻana ʻana, a ua kūkulu ʻia ka uhi o ka paipu mālama. E hoʻohālikelike i ka lua liʻiliʻi me ka piko o ka lima a hoʻopaʻa i ka uhi o ka paipu. Ke hoʻohālikelike nei i nā ʻano ʻelua, palekana ka mea hope. Ke hoʻohana ʻia ka swab haki me kahi paipu mālama liʻiliʻi, hiki ke hoʻoulu i ka wai e pīpī ana i loko o ka paipu i ka wā e haki ai, a pono e uku piha ʻia ka makaʻala o ka haumia i hoʻokumu ʻia e ka hoʻohana pono ʻole ʻana o ka huahana. Manaʻo ʻia e hoʻohana i ka paipu extruded polystyrene (PS) hollow a i ʻole ka polypropylene (PP) injection creasing tube no ka mea o ka lima swab. ʻAʻohe mea i hoʻohana ʻia, ʻaʻole hiki ke hoʻohui ʻia nā mea hoʻohui calcium alginate; nā lāʻau lāʻau a i ʻole nā lāʻau ʻohe. I ka pōkole, pono e hōʻoia ka swab laʻana i ka nui o ka laʻana a me ka nui o ka hoʻokuʻu ʻana, a ʻaʻole pono nā mea i koho ʻia i nā mea e hoʻopilikia i ka hoʻāʻo ʻana ma hope.
2. Hoʻonā mālama virus: ʻElua ʻano o nā hoʻonā mālama virus i hoʻohana nui ʻia ma ka mākeke, ʻo kekahi he hoʻonā mālama virus i hoʻololi ʻia ma muli o ke ʻano halihali, a ʻo kekahi he hoʻonā i hoʻololi ʻia no ka lysate unuhi waikawa nucleic.
ʻO ka ʻāpana nui o ka mea mua, ʻo ia ka medium moʻomeheu kumu a Eagle (MEM) a i ʻole ka paʻakai kaulike a Hank, i hoʻohui ʻia me nā paʻakai, nā waikawa amino, nā huaora, ka glucose a me ka protein e pono ai no ke ola ʻana o ka maʻi. Hoʻohana kēia hopena mālama i ka paʻakai sodium ʻulaʻula phenol ma ke ʻano he hōʻailona a me ka hopena. Ke 6.6-8.0 ka waiwai pH, he poni ka hopena. Hoʻohui ʻia ka glucose, L-glutamine a me ka protein e pono ai i ka hopena mālama. Hāʻawi ʻia ka protein ma ke ʻano o ka serum bovine fetal a i ʻole ka albumin serum bovine, hiki ke hoʻopaʻa i ka pūpū protein o ka maʻi. No ka mea, he waiwai nui ka hopena mālama i nā meaʻai, he mea kūpono ia i ke ola ʻana o ka maʻi akā he mea maikaʻi nō hoʻi i ka ulu ʻana o ka bacteria. Inā haumia ka hopena mālama me ka bacteria, e hoʻonui nui ʻia. ʻO ke kalapona dioxide i loko o kāna metabolites e hoʻoulu ai i ka pH o ka hopena mālama e hāʻule mai ka poni. Huli i ka melemele. No laila, ua hoʻohui ka hapa nui o nā mea hana i nā mea antibacterial i kā lākou mau ʻano. ʻO nā mea antibacterial i ʻōlelo ʻia he penicillin, streptomycin, gentamicin a me polymyxin B. ʻAʻole ʻōlelo ʻia ʻo Sodium azide a me 2-methyl. ʻO nā mea hoʻopale e like me 4-methyl-4-isothiazolin-3-one (MCI) a me 5-chloro-2-methyl-4-isothiazolin-3-one (CMCI) no ka mea he hopena kēia mau ʻāpana i ka hopena PCR. ʻOiai ʻo ka laʻana i hāʻawi ʻia e kēia hopena mālama he virus ola ia, hiki ke mālama ʻia ke ʻano kumu o ka laʻana i ka nui loa, a hiki ke hoʻohana ʻia ʻaʻole wale no ka unuhi ʻana a me ka ʻike ʻana i nā waikawa nucleic virus, akā no ka mahi ʻana a me ka hoʻokaʻawale ʻana i nā virus. Eia nō naʻe, pono e hoʻomaopopo ʻia i ka wā e hoʻohana ʻia ai no ka ʻike ʻana, pono e hana ʻia ka unuhi ʻana a me ka hoʻomaʻemaʻe ʻana o ka waikawa nucleic ma hope o ka hoʻopau ʻana.
ʻO kekahi ʻano hopena mālama i hoʻomākaukau ʻia ma muli o ka lysate unuhi waikawa nucleic, ʻo nā mea nui he mau paʻakai kaulike, EDTA chelating agent, guanidine salt (e like me guanidine isothiocyanate, guanidine hydrochloride, etc.), anionic surfactant (e like me dodecane Sodium sulfate), cationic surfactants (e like me tetradecyltrimethylammonium oxalate), phenol, 8-hydroxyquinoline, dithiothreitol (DTT), proteinase K a me nā mea ʻē aʻe, ʻO kēia hopena mālama ʻana e ʻoki pololei i ka maʻi e hoʻokuʻu i ka waikawa nucleic a hoʻopau i ka RNase. Inā hoʻohana wale ʻia no RT-PCR, ʻoi aku ka kūpono, akā hiki i ka lysate ke hoʻopau i ka maʻi. ʻAʻole hiki ke hoʻohana ʻia kēia ʻano laʻana no ka hoʻokaʻawale ʻana i ka moʻomeheu virus.
ʻŌlelo ʻia e hoʻohana i nā paʻakai EDTA (e like me ka dipotassium ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid, a pēlā aku) no ka mea hoʻopili i ka ion metala i hoʻohana ʻia i loko o ka hopena mālama virus, a ʻaʻole ʻōlelo ʻia e hoʻohana i ka heparin (e like me ka sodium heparin, lithium heparin), i ʻole e hoʻopilikia i ka ʻike ʻana o ka PCR.
3. Paipu mālama: Pono e koho pono ʻia ka mea o ka paipu mālama. Aia nā ʻikepili e hōʻike ana he pili ka polypropylene (Polypropylene) i ka adsorption o ka nucleic acid, ʻoi aku hoʻi ma ka nui o ka ion tension, ʻoi aku ka makemake o ka polyethylene (Polyethylene) ma mua o ka polypropylene (Polypropylene). Maʻalahi e hopu i ka DNA/RNA. ʻOi aku ka kūpono o ka plastic polyethylene-propylene polymer (Polyallomer) a me kekahi mau pahu plastic polypropylene (Polypropylene) i hana kūikawā ʻia no ka mālama ʻana i ka DNA/RNA. Eia kekahi, i ka wā e hoʻohana ai i kahi swab haki, pono e hoʻāʻo ka paipu mālama e koho i kahi pahu me ke kiʻekiʻe ma mua o 8 cm e pale aku i nā mea o loko mai ka pīpī ʻana a me ka haumia i ka wā e haki ai ka swab.
4. Wai no ka hoʻonā mālama hana: Pono e kānana ʻia ka wai ultrapure i hoʻohana ʻia no ka hoʻonā mālama hana ma o kahi membrane ultrafiltration me ke kaumaha molekala o 13,000 e hōʻoia i ka wehe ʻana o nā haumia polymer mai nā kumu olaola, e like me RNase, DNase, a me endotoxin, a ʻaʻole ʻōlelo ʻia ka hoʻomaʻemaʻe maʻamau. Wai a wai i hoʻoheheʻe ʻia paha.
2. Ka hoʻohana ʻana i nā ʻōmole laʻana virus
Ua māhele nui ʻia ka laʻana me ka hoʻohana ʻana i ka ʻōmole laʻana virus i ka laʻana oropharyngeal a me ka laʻana nasopharyngeal:
1. Ka lawe ʻana i ka laʻana oropharyngeal: E kaomi mua i ke alelo me ka mea hoʻohaʻahaʻa alelo, a laila e hoʻolōʻihi i ke poʻo o ka swab laʻana i loko o ka puʻu e holoi i nā tonsils pharyngeal bilateral a me ka paia pharyngeal hope, a holoi i ka paia pharyngeal hope me ka ikaika māmā, e pale i ka hoʻopā ʻana i ka ʻāpana alelo.
2. Laʻana nasopharyngeal: e ana i ka mamao mai ka piko o ka ihu a i ka pepeiao pepeiao me kahi swab a kaha me ka manamana lima, e hoʻokomo i ka swab laʻana i loko o ka lua ihu ma ke kuhikuhi o ka ihu kū pololei (maka), pono e hoʻolōʻihi ka swab ma ka liʻiliʻi o ka hapalua o ka lōʻihi o ka pepeiao pepeiao a hiki i ka piko o ka ihu, e waiho i ka swab i loko o ka ihu no 15-30 kekona, e hoʻohuli mālie i 3-5 mau manawa, a huki i ka swab.
ʻAʻole paʻakikī ke ʻike ʻia mai ke ʻano o ka hoʻohana ʻana, inā he swab oropharyngeal a i ʻole he swab nasopharyngeal, he hana loea ka laʻana, he paʻakikī a haumia. Pili pono ka maikaʻi o ka laʻana i hōʻiliʻili ʻia i ka ʻike ʻana ma hope. Inā he viral load ka laʻana i hōʻiliʻili ʻia, haʻahaʻa, maʻalahi ke hana i nā hopena wahaheʻe, paʻakikī ke hōʻoia i ka maʻi.
Ka manawa hoʻouna: Iune-21-2020
