1. Za kupanga machubu otengera ma virus
Machubu otengera ma virus ndi a zida zamankhwala. Ambiri opanga zapakhomo amalembedwa molingana ndi zinthu zamtundu woyamba, ndipo makampani ochepa amalembedwa molingana ndi katundu wachiwiri. Posachedwapa, kuti akwaniritse zosowa zadzidzidzi za Wuhan ndi malo ena, makampani ambiri atenga "njira yadzidzidzi" "Lembani chilolezo cholembera kalasi yoyamba. Chubu choyezera ma virus chimapangidwa ndi sampling swab, njira yosungira ma virus komanso zoyika zakunja. Popeza palibe mulingo wogwirizana wa dziko kapena muyeso wamakampani, zopangidwa ndi opanga osiyanasiyana zimasiyana kwambiri.
1. Sampling swab: Sampling swab imalumikizana mwachindunji ndi malo opangira sampuli, ndipo zinthu za mutu wa sampuli zimagwirizana kwambiri ndi zomwe zimadziwika. Mutu wa swab wa zitsanzo uyenera kupangidwa ndi ulusi wa Polyester (PE) kapena Rayon (ulusi wopangidwa ndi anthu). Siponji ya calcium alginate kapena swabs zamatabwa (kuphatikiza ndodo zansungwi) sizingagwiritsidwe ntchito, ndipo zida zamutu wa swab sizingakhale za thonje. Chifukwa ulusi wa thonje umakhala ndi mphamvu yolimbikitsira mapuloteni, sikophweka kulowa mu njira yosungiramo; ndipo pamene ndodo yamatabwa kapena nsungwi yomwe ili ndi kashiamu alginate ndi zigawo zamatabwa zathyoledwa, kulowetsedwa mu njira yosungiramo kumapangitsanso mapuloteni, ndipo ngakhale kutero Ikhoza kulepheretsa zotsatira za PCR. Ndibwino kugwiritsa ntchito ulusi wopangidwa monga PE fiber, polyester fiber ndi polypropylene fiber pazinthu zamutu wa swab. Ulusi wachilengedwe monga thonje ndiwosavomerezeka. Ulusi wa nayiloni nawonso ndiwosavomerezeka chifukwa ulusi wa nayiloni (wofanana ndi mitu ya mswaki) umayamwa madzi. Zosakwanira, zomwe zimabweretsa kusakwanira kwa sampuli, zomwe zimakhudza kuchuluka kwa kuzindikira. Siponji ya calcium alginate ndiyoletsedwa kutengera zinthu za swab! Chogwirira cha swab chili ndi mitundu iwiri: yosweka komanso yomangidwa. Nsalu yosweka imayikidwa mu chubu chosungirako pambuyo pa sampuli, ndipo kapu ya chubu imathyoledwa pambuyo posweka kuchokera pamalo pafupi ndi mutu wa chitsanzo; swab yomangidwa mwachindunji imayika sampuli mu chubu chosungirako pambuyo pa zitsanzo, ndipo chivundikiro cha chubu chosungira chimamangidwa mu Gwirizanitsani kabowo kakang'ono ndi pamwamba pa chogwirira ndikumangitsa chivundikiro cha chubu. Poyerekeza njira ziwirizi, yotsirizirayi ndi yotetezeka. Pamene swab yosweka imagwiritsidwa ntchito limodzi ndi kachubu kakang'ono kakusungirako, kungayambitse kuphulika kwamadzi mu chubu pamene kusweka, ndipo chidwi chonse chiyenera kuperekedwa ku chiopsezo cha kuipitsidwa chifukwa chogwiritsa ntchito molakwika mankhwala. Ndi bwino kugwiritsa ntchito dzenje polystyrene (PS) extruded chubu kapena polypropylene (PP) jekeseni creasing chubu kwa zinthu za swab chogwirira. Ziribe kanthu zomwe zimagwiritsidwa ntchito, zowonjezera za calcium alginate sizikhoza kuwonjezeredwa; matabwa kapena nsungwi. Mwachidule, sampling swab iyenera kutsimikizira kuchuluka kwa sampuli ndi kuchuluka kwa kutulutsidwa, ndipo zida zosankhidwa siziyenera kukhala ndi zinthu zomwe zimakhudza kuyezetsa kotsatira.
2. Virus kuteteza njira: Pali mitundu iwiri ya HIV kuteteza njira chimagwiritsidwa ntchito pamsika, imodzi ndi HIV yokonza njira kusinthidwa kutengera sing'anga zoyendera, ndipo ina ndi kusinthidwa njira kwa nucleic asidi m'zigawo lysate.
Gawo lalikulu lakale ndi Eagle's basic culture medium (MEM) kapena mchere wa Hank, womwe umawonjezeredwa ndi mchere, ma amino acid, mavitamini, shuga ndi mapuloteni ofunikira kuti ma virus apulumuke. Njira yosungirayi imagwiritsa ntchito mchere wa phenol wofiira wa sodium monga chizindikiro ndi yankho. Pamene pH mtengo ndi 6.6-8.0, yankho ndi pinki. Glucose wofunikira, L-glutamine ndi mapuloteni amawonjezeredwa ku njira yosungira. Puloteni amaperekedwa mu mawonekedwe a fetal bovine seramu kapena bovine seramu albumin, amene angathe kukhazikika mapuloteni chipolopolo cha HIV. Chifukwa chakuti njira yotetezera imakhala ndi michere yambiri, imathandizira kupulumuka kwa kachilomboka komanso imapindulitsa pakukula kwa mabakiteriya. Ngati njira yosungirayi ili ndi mabakiteriya, idzachulukana mochuluka. Mpweya woipa wa carbon dioxide m'ma metabolites ake udzachititsa kuti pH yosungiramo igwe kuchokera ku pinki Imasanduka chikasu. Chifukwa chake, opanga ambiri adawonjezera zosakaniza za antibacterial pakupanga kwawo. Mankhwala oletsa tizilombo toyambitsa matenda ndi penicillin, streptomycin, gentamicin ndi polymyxin B. Sodium azide ndi 2-methyl sakuvomerezeka Zoletsa monga 4-methyl-4-isothiazolin-3-one (MCI) ndi 5-chloro-2-methyl-4. -isothiazolin-3-one (CMCI) chifukwa zigawozi zimakhala ndi zotsatira za PCR. Popeza chitsanzo choperekedwa ndi njira yotetezerayi ndi kachilombo koyambitsa matenda, chiyambi cha chitsanzocho chikhoza kusungidwa kwambiri, ndipo chingagwiritsidwe ntchito osati kungochotsa ndi kuzindikira kachilombo ka nucleic acids, komanso kulima ndi kulima. kudzipatula kwa ma virus. Komabe, ziyenera kudziwidwa kuti zikagwiritsidwa ntchito pozindikira, kutulutsa kwa nucleic acid ndi kuyeretsa kuyenera kuchitidwa pambuyo poyambitsa.
Mtundu wina wa njira kuteteza anakonza zochokera nucleic asidi m'zigawo lysate, zigawo zikuluzikulu ndi bwino mchere, EDTA chelating wothandizira, guanidine mchere (monga guanidine isothiocyanate, guanidine hydrochloride, etc.), anionic surfactant (monga dodecane Sodium sulfate), surfactants (monga tetradecyltrimethylammonium oxalate), phenol, 8-hydroxyquinoline, dithiothreitol (DTT), proteinase K ndi zigawo zina, Njira yosungirayi ndiyo kuchotsa mwachindunji kachilomboka kuti amasule nucleic acid ndikuchotsa RNase. Ngati ntchito RT-PCR, ndi abwino kwambiri, koma lysate akhoza inactivate kachilombo. Zitsanzo zamtunduwu sizingagwiritsidwe ntchito pakulekanitsa chikhalidwe cha ma virus.
The zitsulo ion chelating wothandizila ntchito HIV kuteteza tizilombo tikulimbikitsidwa kugwiritsa ntchito mchere EDTA (monga dipotassium ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic asidi, etc.), ndipo si bwino kugwiritsa ntchito heparin (monga sodium heparin, lithiamu heparin), kuti asakhudze kuzindikira kwa PCR.
3. Chubu chosungira: Zinthu za chubu chosungira ziyenera kusankhidwa mosamala. Pali deta yomwe imasonyeza kuti polypropylene (Polypropylene) imagwirizana ndi kutsekemera kwa nucleic acid, makamaka pamagulu othamanga kwambiri a ion, polyethylene (Polyethylene) ndi yabwino kuposa polypropylene (Polypropylene) Yosavuta kumvetsa DNA/RNA. Mapulasitiki a polyethylene-propylene polima (Polyallomer) ndi zotengera zapulasitiki za polypropylene (Polypropylene) ndizoyenera kusungirako DNA/RNA. Kuonjezera apo, pogwiritsira ntchito swab yosweka, chubu yosungiramo iyenera kuyesa kusankha chidebe chokhala ndi msinkhu woposa 8 cm kuti zomwe zili mkati zisawonongeke ndi kuipitsidwa pamene swab yathyoka.
4. Madzi opangira njira yotetezera kupanga: Madzi amtundu wa ultrapure omwe amagwiritsidwa ntchito posungirako madzi ayenera kusefedwa kudzera mu ultrafiltration nembanemba yokhala ndi molekyulu yolemera 13,000 kuti atsimikizire kuchotsa zonyansa za polima kuchokera kuzinthu zachilengedwe, monga RNase, DNase, ndi endotoxin, ndi kuyeretsedwa wamba sikuvomerezeka. Madzi kapena madzi osungunuka.
2. Kugwiritsa ntchito machubu oyesa ma virus
Sampling pogwiritsa ntchito ma virus sampling chubu makamaka amagawidwa mu oropharyngeal sampling ndi nasopharyngeal sampling:
1. Oropharyngeal sampling: Choyamba kanikizani lilime ndi depressor lilime, kenaka tambasulani mutu wa chitsanzo swab ku mmero kuti mupukute mbali ziwiri za pharyngeal tonsils ndi posterior pharyngeal khoma, ndi kupukuta pambuyo pharyngeal khoma ndi kuwala mphamvu, kupewa kukhudza lilime. unit.
2. Sampuli ya Nasopharyngeal: kuyeza mtunda kuchokera kunsonga ya mphuno kupita ku nsonga ya khutu ndi swab ndikuyika chizindikiro ndi chala, ikani chitsanzocho mumphuno yamphuno molunjika mphuno yowongoka (nkhope), swab iyenera kufalikira. osachepera theka la kutalika kwa khutu mpaka nsonga ya mphuno, Siyani swab m'mphuno kwa masekondi 15-30, tembenuzani pang'onopang'ono. 3-5 nthawi, ndikuchotsa swab.
Sizovuta kuwona kuchokera ku njira yogwiritsira ntchito, kaya ndi swab ya oropharyngeal kapena swab ya nasopharyngeal, sampuli ndi ntchito yaukadaulo, yomwe imakhala yovuta komanso yoipitsidwa. Ubwino wa chitsanzo chosonkhanitsidwa umagwirizana mwachindunji ndi kudziwika kotsatira. Ngati chitsanzo chosonkhanitsidwa chili ndi ma virus otsika, osavuta kuyambitsa zoyipa zabodza, zovuta kutsimikizira matenda.
Nthawi yotumiza: Jun-21-2020
