Ob peb lub tswv yim ntawm cov raj kuaj kab mob

1. Txog kev tsim cov raj kuaj kab mob
Cov raj kuaj kab mob yog cov khoom siv kho mob. Feem ntau cov chaw tsim khoom hauv tsev tau sau npe raws li cov khoom lag luam zoo tshaj plaws, thiab ob peb lub tuam txhab tau sau npe raws li cov khoom lag luam zoo tshaj plaws. Tsis ntev los no, txhawm rau kom tau raws li qhov xav tau xwm txheej ceev ntawm Wuhan thiab lwm qhov chaw, ntau lub tuam txhab tau siv "kev xa xov xwm ceev" "Thov rau daim ntawv tso cai sau npe zoo tshaj plaws." Lub raj kuaj kab mob yog tsim los ntawm cov ntaub so ntswg kuaj, cov tshuaj tiv thaiv kab mob thiab cov ntim sab nraud. Vim tias tsis muaj tus qauv hauv tebchaws lossis tus qauv kev lag luam sib koom ua ke, cov khoom lag luam ntawm ntau lub chaw tsim khoom sib txawv heev.

1. Lub swab kuaj: Lub swab kuaj ncaj qha kov qhov chaw kuaj, thiab cov khoom siv ntawm lub taub hau kuaj muaj feem cuam tshuam nrog kev kuaj tom qab. Lub taub hau swab kuaj yuav tsum ua los ntawm Polyester (PE) cov fiber hluavtaws lossis Rayon (cov fiber ua los ntawm tib neeg). Calcium alginate daim txhuam cev lossis cov ntoo pas nrig swabs (xws li cov pas nrig xyoob) tsis tuaj yeem siv, thiab cov khoom siv ntawm lub taub hau swab tsis tuaj yeem ua cov khoom paj rwb. Vim tias cov paj rwb fiber muaj zog nqus cov protein, nws tsis yooj yim rau elut rau hauv cov tshuaj khaws cia tom qab; thiab thaum ib tug pas nrig ntoo lossis cov ntoo pas nrig uas muaj calcium alginate thiab cov khoom ntoo raug tawg, kev tsau rau hauv cov tshuaj khaws cia kuj tseem yuav nqus cov protein, thiab txawm tias nws tuaj yeem tiv thaiv PCR cov tshuaj tiv thaiv tom qab. Nws raug pom zoo kom siv cov fiber hluavtaws xws li PE fiber, polyester fiber thiab polypropylene fiber rau cov khoom siv ntawm lub taub hau swab. Cov fiber ntuj xws li paj rwb tsis pom zoo. Cov fiber nylon kuj tsis pom zoo vim tias cov fiber nylon (zoo ib yam li cov taub hau txhuam hniav) nqus dej. Phem, ua rau muaj qhov ntim kuaj tsis txaus, cuam tshuam rau qhov nrawm ntawm kev kuaj. Calcium alginate daim txhuam cev raug txwv tsis pub siv rau cov khoom siv swab kuaj! Lub tes tuav swab muaj ob hom: tawg thiab ua tiav. Tom qab kuaj cov ntaub so ntswg uas tawg lawm, muab tso rau hauv lub raj xa khoom, thiab lub hau raj raug tawg tom qab raug tawg los ntawm qhov chaw ze ntawm lub taub hau kuaj; lub ntaub so ntswg uas ua rau hauv muab cov ntaub so ntswg kuaj ncaj qha rau hauv lub raj xa khoom tom qab kuaj, thiab lub hau raj xa khoom raug tsim. Kho lub qhov me me nrog rau sab saum toj ntawm tus kov thiab nruj lub hau raj. Piv rau ob txoj kev, qhov tom kawg muaj kev nyab xeeb heev. Thaum siv cov ntaub so ntswg uas tawg lawm ua ke nrog lub raj xa khoom me dua, nws yuav ua rau cov kua dej ntws hauv lub raj thaum tawg, thiab yuav tsum tau them sai sai rau qhov kev pheej hmoo ntawm kev ua qias tuaj los ntawm kev siv cov khoom tsis raug. Nws raug pom zoo kom siv cov raj polystyrene hollow (PS) extruded lossis polypropylene (PP) txhaj tshuaj creasing rau cov khoom siv ntawm tus kov. Txawm hais tias siv cov khoom siv dab tsi los xij, cov khoom ntxiv calcium alginate tsis tuaj yeem ntxiv; cov pas ntoo lossis cov pas xyoob. Hauv ntej, cov ntaub so ntswg kuaj yuav tsum xyuas kom meej tias muaj pes tsawg cov qauv thiab qhov ntau ntawm kev tso tawm, thiab cov khoom xaiv yuav tsum tsis muaj cov tshuaj uas cuam tshuam rau kev sim tom qab.

2. Kev daws teeb meem tiv thaiv kab mob: Muaj ob hom kev daws teeb meem tiv thaiv kab mob uas siv dav hauv kev ua lag luam, ib qho yog kev daws teeb meem tiv thaiv kab mob hloov kho raws li qhov nruab nrab thauj mus los, thiab lwm qhov yog kev daws teeb meem hloov kho rau nucleic acid extraction lysate.
Cov khoom tseem ceeb ntawm cov qub yog Eagle's basic culture medium (MEM) lossis Hank's balanced salt, uas ntxiv nrog cov ntsev, amino acids, vitamins, glucose thiab protein tsim nyog rau kev ciaj sia ntawm tus kab mob. Cov tshuaj khaws cia no siv phenol liab sodium ntsev ua tus qhia thiab cov tshuaj. Thaum tus nqi pH yog 6.6-8.0, cov tshuaj yog xim liab. Cov glucose, L-glutamine thiab protein tsim nyog raug ntxiv rau cov tshuaj khaws cia. Cov protein tau muab rau hauv daim ntawv ntawm fetal bovine serum lossis bovine serum albumin, uas tuaj yeem ua kom lub plhaub protein ntawm tus kab mob ruaj khov. Vim tias cov tshuaj khaws cia muaj ntau cov as-ham, nws yog qhov zoo rau kev ciaj sia ntawm tus kab mob tab sis kuj pab rau kev loj hlob ntawm cov kab mob. Yog tias cov tshuaj khaws cia muaj cov kab mob, nws yuav nce ntau heev. Cov pa roj carbon dioxide hauv nws cov metabolites yuav ua rau cov tshuaj khaws cia pH poob qis los ntawm xim liab tig daj. Yog li ntawd, feem ntau cov chaw tsim khoom tau ntxiv cov khoom xyaw antibacterial rau lawv cov qauv. Cov tshuaj tua kab mob uas pom zoo yog penicillin, streptomycin, gentamicin thiab polymyxin B. Sodium azide thiab 2-methyl tsis pom zoo. Cov tshuaj Inhibitors xws li 4-methyl-4-isothiazolin-3-one (MCI) thiab 5-chloro-2-methyl-4-isothiazolin-3-one (CMCI) vim tias cov khoom no muaj feem cuam tshuam rau PCR reaction. Txij li thaum cov qauv uas muab los ntawm cov tshuaj khaws cia no yog ib hom kab mob uas muaj sia nyob, qhov qub ntawm cov qauv tuaj yeem khaws cia rau qhov loj tshaj plaws, thiab nws tuaj yeem siv tsis yog rau kev rho tawm thiab nrhiav pom cov kab mob nucleic acids xwb, tab sis kuj rau kev cog qoob loo thiab kev cais tawm ntawm cov kab mob. Txawm li cas los xij, nws yuav tsum tau sau tseg tias thaum siv rau kev nrhiav pom, kev rho tawm thiab kev ntxuav nucleic acid yuav tsum tau ua tom qab tsis ua haujlwm.
Lwm hom kev khaws cia uas tau npaj raws li nucleic acid extraction lysate, cov khoom tseem ceeb yog cov ntsev sib npaug, EDTA chelating agent, guanidine ntsev (xws li guanidine isothiocyanate, guanidine hydrochloride, thiab lwm yam), anionic surfactant (xws li dodecane Sodium sulfate), cationic surfactants (xws li tetradecyltrimethylammonium oxalate), phenol, 8-hydroxyquinoline, dithiothreitol (DTT), proteinase K thiab lwm yam khoom. Cov tshuaj khaws cia no yog los txiav cov kab mob ncaj qha kom tso cov nucleic acid thiab tshem tawm RNase. Yog tias tsuas yog siv rau RT-PCR, nws tsim nyog dua, tab sis lysate tuaj yeem ua rau cov kab mob tsis ua haujlwm. Hom qauv no tsis tuaj yeem siv rau kev sib cais kab mob.

Tus neeg sawv cev hlau ion chelating siv rau hauv cov tshuaj tiv thaiv kab mob raug pom zoo kom siv cov ntsev EDTA (xws li dipotassium ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid, thiab lwm yam), thiab nws tsis pom zoo kom siv heparin (xws li sodium heparin, lithium heparin), yog li ntawd kom tsis txhob cuam tshuam rau PCR kev kuaj pom.
3. Lub raj mis khaws cia: Cov khoom siv ntawm lub raj mis khaws cia yuav tsum tau xaiv kom zoo. Muaj cov ntaub ntawv qhia tias polypropylene (Polypropylene) muaj feem cuam tshuam nrog kev nqus ntawm nucleic acid, tshwj xeeb tshaj yog ntawm qhov siab ntawm cov ion concentration, polyethylene (Polyethylene) yog qhov nyiam dua li polypropylene (Polypropylene). Yooj yim rau kev nkag siab DNA / RNA. Polyethylene-propylene polymer (Polyallomer) yas thiab qee cov thawv yas polypropylene (Polypropylene) uas tau ua tshwj xeeb yog qhov tsim nyog dua rau kev khaws cia DNA / RNA. Tsis tas li ntawd, thaum siv lub swab uas tawg tau, lub raj mis khaws cia yuav tsum sim xaiv lub thawv uas muaj qhov siab tshaj 8 cm kom tiv thaiv cov ntsiab lus los ntawm kev txaws thiab ua qias tuaj thaum lub swab tawg.

4. Dej rau kev khaws cia cov tshuaj: Cov dej ntshiab heev uas siv rau kev khaws cia cov tshuaj yuav tsum tau lim los ntawm daim nyias nyias ultrafiltration nrog qhov hnyav molecular ntawm 13,000 kom ntseeg tau tias tshem tawm cov polymer impurities los ntawm cov chaw muaj sia, xws li RNase, DNase, thiab endotoxin, thiab tsis pom zoo kom ntxuav li niaj zaus. Dej lossis dej distilled.

2. Siv cov raj kuaj kab mob

Kev kuaj siv lub raj kuaj kab mob feem ntau yog muab faib ua kev kuaj oropharyngeal thiab kev kuaj nasopharyngeal:

1. Kev kuaj mob ntawm lub qhov ncauj thiab caj pas: Ua ntej nias tus nplaig nrog lub tshuab nias tus nplaig, tom qab ntawd nthuav lub taub hau ntawm daim ntaub so quav mus rau hauv caj pas kom so ob sab caj pas thiab phab ntsa caj pas tom qab, thiab so phab ntsa caj pas tom qab nrog lub zog me me, tsis txhob kov lub qhov ncauj.

2. Kev kuaj nasopharyngeal: ntsuas qhov deb ntawm qhov ntswg mus rau pob ntseg nrog ib daim ntaub so ntswg thiab cim nrog ib tug ntiv tes, ntxig daim ntaub so ntswg rau hauv qhov ntswg raws li qhov ntswg ntsug (lub ntsej muag), daim ntaub so ntswg yuav tsum ntev tsawg kawg ib nrab ntawm qhov ntev ntawm pob ntseg mus rau qhov ntswg. Cia daim ntaub so ntswg nyob hauv qhov ntswg li 15-30 vib nas this, maj mam tig 3-5 zaug, thiab rho daim ntaub so ntswg tawm.
Los ntawm txoj kev siv, txawm tias nws yog daim ntaub so ntswg oropharyngeal lossis daim ntaub so ntswg nasopharyngeal, kev kuaj mob yog ib txoj haujlwm txuj ci, uas nyuaj thiab muaj kuab paug. Qhov zoo ntawm cov qauv sau tau muaj feem cuam tshuam ncaj qha rau kev kuaj pom tom qab. Yog tias cov qauv sau tau muaj tus kab mob tsawg, yooj yim ua rau muaj qhov tsis zoo cuav, nyuaj rau paub meej tias kuaj mob.