1. Zokhudza kupanga machubu oyesera ma virus
Machubu otengera ma virus ndi a zipangizo zachipatala. Opanga ambiri m'nyumba amalembetsa malinga ndi zinthu zapamwamba, ndipo makampani ochepa amalembetsa malinga ndi zinthu zapamwamba. Posachedwapa, kuti akwaniritse zosowa zadzidzidzi ku Wuhan ndi madera ena, makampani ambiri atenga "njira yodzidzimutsa" "Lembani chilolezo cholembera ma virus apamwamba. Chubu chotengera ma virus chimapangidwa ndi swab yotsanzira, yankho losungira ma virus ndi phukusi lakunja. Popeza palibe muyezo umodzi wadziko lonse kapena muyezo wamakampani, zinthu za opanga osiyanasiyana zimasiyana kwambiri.
1. Chisambitso cha zitsanzo: Chisambitso cha zitsanzo chimalumikizana mwachindunji ndi malo oyesera, ndipo zinthu za mutu wa chitsanzo zimagwirizana kwambiri ndi kuzindikirika komwe kukubwera. Mutu wa chisambitso cha zitsanzo uyenera kupangidwa ndi ulusi wopangidwa ndi Polyester (PE) kapena Rayon (ulusi wopangidwa ndi munthu). Siponji ya calcium alginate kapena ziboliboli zamatabwa (kuphatikiza ndodo za nsungwi) sizingagwiritsidwe ntchito, ndipo zinthu za mutu wa chisambitso sizingakhale zopangidwa ndi thonje. Chifukwa ulusi wa thonje uli ndi mayamwidwe amphamvu a mapuloteni, sizophweka kulowa mu yankho losungira lotsatira; ndipo ndodo yamatabwa kapena ndodo ya nsungwi yokhala ndi calcium alginate ndi zigawo zamatabwa zikasweka, kuviika mu yankho losungira kudzayamwitsanso mapuloteni, ndipo ngakhale zidzaletsa PCR reaction yotsatira. Ndikoyenera kugwiritsa ntchito ulusi wopangidwa monga ulusi wa PE, ulusi wa polyester ndi ulusi wa polypropylene pazinthu za mutu wa chisambitso. Ulusi wachilengedwe monga thonje sukulimbikitsidwa. Ulusi wa nayiloni sukulimbikitsidwanso chifukwa ulusi wa nayiloni (wofanana ndi mitu ya burashi ya mano) umayamwa madzi. Zoipa, zomwe zimapangitsa kuti kuchuluka kwa zitsanzo kusakwane, zomwe zimakhudza kuchuluka kwa kuzindikira. Chisambitso cha calcium alginate sichiloledwa pazinthu zoyesera! Chogwirira cha swab chili ndi mitundu iwiri: chosweka ndi chomangidwa mkati. Swab yosweka imayikidwa mu chubu chosungiramo zinthu pambuyo poyesa, ndipo chivundikiro cha chubu chimasweka pambuyo posweka kuchokera pamalo omwe ali pafupi ndi mutu wa chitsanzo; swab yomangidwa mkati imayika mwachindunji swab yoyesera mu chubu chosungiramo zinthu pambuyo poyesa, ndipo chivundikiro cha chubu chosungiramo zinthu chimamangidwa mkati. Lumikizani dzenje laling'ono ndi pamwamba pa chogwirira ndikulimbitsa chivundikiro cha chubu. Poyerekeza njira ziwirizi, njira yomalizayi ndi yotetezeka. Ngati swab yosweka ikugwiritsidwa ntchito limodzi ndi chubu chosungiramo zinthu chaching'ono, ingayambitse kufalikira kwa madzi mu chubu ikasweka, ndipo chisamaliro chonse chiyenera kuperekedwa ku chiopsezo cha kuipitsidwa komwe kumachitika chifukwa chogwiritsa ntchito molakwika mankhwalawa. Ndikoyenera kugwiritsa ntchito chubu chopanda kanthu cha polystyrene (PS) kapena chubu choboola cha polypropylene (PP) pazinthu za chogwirira cha swab. Kaya mugwiritsa ntchito zinthu ziti, zowonjezera za calcium alginate sizingawonjezedwe; ndodo zamatabwa kapena ndodo za Bamboo. Mwachidule, swab yoyesera iyenera kuwonetsetsa kuchuluka kwa zitsanzo ndi kuchuluka kwa kutulutsidwa, ndipo zipangizo zomwe zasankhidwa siziyenera kukhala ndi zinthu zomwe zimakhudza mayeso otsatira.
2. Njira yosungira ma virus: Pali mitundu iwiri ya njira zosungira ma virus zomwe zimagwiritsidwa ntchito kwambiri pamsika, imodzi ndi njira yosungira ma virus yomwe yasinthidwa kutengera njira yoyendera, ndipo inayo ndi njira yosinthira ya nucleic acid extraction lysate.
Gawo lalikulu la yoyamba ndi Eagle's basic culture medium (MEM) kapena Hank's balanced salt, yomwe imawonjezedwa ndi mchere, ma amino acid, mavitamini, shuga ndi mapuloteni ofunikira kuti kachilombo kapulumuke. Njira yosungirayi imagwiritsa ntchito mchere wofiira wa phenol ngati chizindikiro ndi yankho. Pamene pH ili 6.6-8.0, yankho limakhala lofiirira. Glucose wofunikira, L-glutamine ndi mapuloteni amawonjezedwa ku yankho losungira. Puloteniyi imaperekedwa mu mawonekedwe a fetal bovine serum kapena bovine serum albumin, yomwe imatha kukhazikika chipolopolo cha mapuloteni a kachilomboka. Chifukwa yankho losungira lili ndi michere yambiri, limathandiza kuti kachilomboka kapulumuke komanso limathandiza kukula kwa mabakiteriya. Ngati yankho losungira lili ndi mabakiteriya, lidzachulukana kwambiri. Mpweya wa carbon dioxide womwe uli m'ma metabolites ake udzapangitsa kuti pH ya yankho losungira igwe kuchokera ku pinki. Chifukwa chake, opanga ambiri awonjezera zosakaniza zotsutsana ndi mabakiteriya ku mapangidwe awo. Mankhwala oletsa mabakiteriya omwe amalimbikitsidwa ndi penicillin, streptomycin, gentamicin ndi polymyxin B. Sodium azide ndi 2-methyl sizikulimbikitsidwa. Zoletsa monga 4-methyl-4-isothiazolin-3-one (MCI) ndi 5-chloro-2-methyl-4-isothiazolin-3-one (CMCI) chifukwa zigawozi zimakhudza momwe PCR imagwirira ntchito. Popeza chitsanzo chomwe chaperekedwa ndi yankho loteteza ili ndi kachilombo kamoyo, chiyambi cha chitsanzocho chikhoza kusungidwa bwino kwambiri, ndipo chingagwiritsidwe ntchito osati pongochotsa ndi kuzindikira ma nucleic acid a kachilombo kokha, komanso polima ndi kusiyanitsa mavairasi. Komabe, ziyenera kudziwika kuti zikagwiritsidwa ntchito pozindikira, kuchotsa ndi kuyeretsa nucleic acid kuyenera kuchitika mutasiya kugwira ntchito.
Mtundu wina wa njira yosungira yokonzedwa kutengera lysate yochotsera nucleic acid, zigawo zazikulu ndi mchere wolinganizidwa, EDTA chelating agent, mchere wa guanidine (monga guanidine isothiocyanate, guanidine hydrochloride, ndi zina zotero), anionic surfactant (monga dodecane Sodium sulfate), cationic surfactants (monga tetradecyltrimethylammonium oxalate), phenol, 8-hydroxyquinoline, dithiothreitol (DTT), proteinase K ndi zigawo zina. Njira yosungirayi ndi yodula mwachindunji kachilomboka kuti itulutse nucleic acid ndikuchotsa RNase. Ngati igwiritsidwa ntchito kokha pa RT-PCR, ndiyoyenera kwambiri, koma lysate imatha kuletsa kachilomboka. Mtundu uwu wa chitsanzo sungagwiritsidwe ntchito polekanitsa kukula kwa kachilomboka.
Chothandizira chelating chachitsulo chomwe chimagwiritsidwa ntchito mu yankho loteteza kachilomboka chikulimbikitsidwa kugwiritsa ntchito mchere wa EDTA (monga dipotassium ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid, ndi zina zotero), ndipo sikoyenera kugwiritsa ntchito heparin (monga sodium heparin, lithium heparin), kuti zisakhudze kupezeka kwa PCR.
3. Chubu chosungira: Zinthu zomwe zili mu chubu chosungira ziyenera kusankhidwa mosamala. Pali deta yomwe ikusonyeza kuti polypropylene (Polypropylene) imagwirizana ndi kuyamwa kwa nucleic acid, makamaka pa kuchuluka kwa ma ion okwera, polyethylene (Polyethylene) ndiyokondedwa kwambiri kuposa polypropylene (Polypropylene). DNA/RNA yosavuta kugwira. Mapulasitiki a polyethylene-propylene polymer (Polyallomer) ndi mapulasitiki ena a polypropylene (Polypropylene) omwe amakonzedwa mwapadera ndi oyenera kusungira DNA/RNA. Kuphatikiza apo, pogwiritsa ntchito swab yosweka, chubu chosungira chiyenera kuyesa kusankha chidebe chokhala ndi kutalika kopitilira 8 cm kuti chiteteze zomwe zili mkati kuti zisatayike ndi kuipitsidwa pamene swab yasweka.
4. Madzi osungira zinthu: Madzi oyera kwambiri omwe amagwiritsidwa ntchito posungira zinthu ayenera kusefedwa kudzera mu nembanemba yoyezera zinthu mopitirira muyeso yokhala ndi kulemera kwa mamolekyulu 13,000 kuti zitsiru za polima zichotsedwe kuchokera kuzinthu zachilengedwe, monga RNase, DNase, ndi endotoxin, ndipo kuyeretsa wamba sikuvomerezeka. Madzi kapena madzi osungunuka.
2. Kugwiritsa ntchito machubu oyesera ma virus
Kusankha zitsanzo pogwiritsa ntchito chubu choyezera kachilombo kumagawidwa makamaka m'magulu awiri: oropharyngeal sampling ndi nasopharyngeal sampling:
1. Kusankha m'kamwa: Choyamba kanikizani lilime ndi choletsa lilime, kenako tambasulani mutu wa swab yosankha m'khosi kuti mupukute matoni a pharyngeal ndi khoma la posterior pharyngeal, ndikupukuta khoma la posterior pharyngeal ndi mphamvu yopepuka, pewani kukhudza lilime.
2. Kuyesa m'mphuno: yesani mtunda kuchokera ku nsonga ya mphuno kupita ku khutu ndi swab ndikuyika chizindikiro ndi chala, ikani swab yoyesera m'mphuno molunjika ku mphuno yoyimirira (nkhope), swab iyenera kufalikira osachepera theka la kutalika kwa khutu mpaka ku nsonga ya mphuno. Siyani swab m'mphuno kwa masekondi 15-30, tembenuzani pang'onopang'ono katatu-kasanu, ndikuchotsa swab.
Sikovuta kuona kuchokera ku njira yogwiritsira ntchito, kaya ndi swab ya oropharyngeal kapena swab ya nasopharyngeal, kutengera zitsanzo ndi ntchito yaukadaulo, yomwe ndi yovuta komanso yodetsedwa. Ubwino wa chitsanzo chosonkhanitsidwa umagwirizana mwachindunji ndi kupezeka pambuyo pake. Ngati chitsanzo chosonkhanitsidwacho chili ndi kuchuluka kwa mavairasi. Zochepa, zosavuta kuyambitsa zoipa zabodza, zovuta kutsimikizira matendawa.
Nthawi yotumizira: Juni-21-2020
