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 ku Wuhan ndi malo ena, makampani ambiri atenga "njira yadzidzidzi" "Pemphani chilolezo cha kalasi yoyamba." Chubu chotengera kachilombo ka HIV chimapangidwa ndi sampling swab, njira yosungira ma virus ndi zoyika zakunja.
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. Ma antibacterial agents omwe amalangizidwa ndi penicillin, streptomycin, gentamicin ndi polymyxin B. Sodium azide ndi 2-methyl sakuvomerezedwa Inhibitors monga 4-methyl-4-isothiazolin-3-one (MCI) ndi 5-chloro-2-methyl-4-isothiazolin-3-imodzi (CMCI) chifukwa zigawozi zili ndi zotsatira za PCR. Popeza chitsanzo choperekedwa ndi njira yotetezerayi ndi kachilombo koyambitsa matenda, chiyambi cha chitsanzocho chikhoza kusungidwa kwambiri, ndipo chingagwiritsidwe ntchito osati pochotsa ndi kuzindikira kachilombo ka nucleic acids, komanso kulima ndi kudzipatula kwa mavairasi. 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 wothandizila, guanidine mchere (monga guanidine isothiocyanate, guanidine hydrochloride, etc.), anionic surfactant (monga dodecane Sodium sulfate sulfate) 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.
Chitsulo cha ion chelating agent chomwe chimagwiritsidwa ntchito mu njira yothetsera kachilombo ka HIV tikulimbikitsidwa kugwiritsa ntchito mchere wa EDTA (monga dipotassium ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid, etc.), ndipo sikulimbikitsidwa 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 mankhwala 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 kuyeretsa 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 lilime depressor, kenaka tambani mutu wa chitsanzo swab kukhosi kuti misozi apawiri pharyngeal tonsils ndi posterior pharyngeal khoma, ndi kupukuta posterior 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 ndikulemba ndi chala, ikani chitsanzo chachitsulo mumphuno yamphuno molunjika mphuno yowongoka (nkhope), swab iyenera kupitirira osachepera theka la kutalika kwa khutu mpaka kunsonga kwa mphuno, Siyani no30 swab yachiwiri, 5. 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
