Tus Neeg Tshaj Lij Coronavirus (SARS-CoV-2) Multiplex Real-Time PCR Cov Khoom Siv

Cov lus piav qhia luv luv:

Tus kabmob tshiab coronavirus yog tus kabmob RNA, uas muaj cov protein thiab nucleic acids. Tus kab mob kis tau tus tswv tsev (tib neeg) lub cev, nkag rau hauv cov hlwb los ntawm qhov chaw sib txuas lub zog ACE2, thiab rov ua haujlwm hauv cov hlwb ua haujlwm, ua rau tib neeg lub cev tiv thaiv kab mob rau teb txawv teb chaws thiab tsim cov tshuaj tiv thaiv tshwj xeeb. Yog li no, lub vial nucleic acids thiab antigens, thiab cov tshuaj tiv thaiv tshwj xeeb tiv thaiv tus kab mob coronavirus tshiab tuaj yeem siv raws li cov biomarkers tshwj xeeb rau kev kuaj pom tus mob tshiab coronavirus. Txog kev tshawb nrhiav nucleic acid, RT-PCR thev naus laus zis feem ntau tau siv.

Novel Coronavirus (SARS-CoV-2) Multiplex Real-Time PCR Cov khoom siv yog xav kom siv los ua kom tau qhov kev ua tau zoo ntawm SARS_CoV-2 viral RNA muab los ntawm nasopharyngeal swabs, oropharyngeal swabs, hnoos qeev thiab BALF los ntawm cov neeg mob koom nrog koom nrog FDA / CE IVD kev rho tawm system thiab cov raug xaiv los ntawm PCR cov npe teev saum toj no.

Cov khoom siv yog npaj rau kev siv los ntawm kev ua haujlwm hauv chav kuaj


Khoom Nthuav dav

Cov khoom lag luam

Qhov kub siab heev, npaj siv PCR cov khoom siv muaj cov ntawv lyophilized (khov-qhuav tsim cov txheej txheem) rau lub sijhawm ntev cia. Cov khoom siv tuaj yeem tuaj yeem thauj thiab khaws cia ntawm chav tsev kub thiab nyob ruaj rau ib xyoos. Txhua lub raj ntawm premix muaj tag nrho cov reagents xav tau rau kev nthuav dav PCR, suav nrog Reverse-transcriptase, Taq polymerase, primers, probes, thiab dNTPs substrates. Nws tsuas yog xav tau ntxiv 13ul dej distilled thiab 5ul muab rho tawm RNA template, tom qab ntawd nws tuaj yeem khiav thiab nthuav dav ntawm PCR cov twj paj nruag.

Lub tshuab qPCR yuav tsum ua tau raws li cov hauv qab no:
1. Ua kom haum 8 pluaj PCR raj ntim 0.2 ml
2. Muaj ntau tshaj plaub qhov chaw nrhiav pom:

Channel

Kev lom zem (nm)

Emission (nm)

Pre-Calibrated Dyes Ua Ntej

1.

470

525

FAM, SYBR Ntsuab I

2

523

564

VIC, HEX, TET, JOE

3.

571

621

ROX, TEXAS-LIAB

4

630

670

CY5

PCR-platform:
7500Qhov Sijhawm-PCR System, Biorad CF96, iCycler iQ ™ Real-Time PCR Kev Tshawb Nrhiav Txheej Txheem, Stratagene Mx3000P, Mx3005P

Kev nyuaj ntawm kev thauj tus mob khaub thuas txias ntawm Novel Coronavirus nucleic acid kuaj pom reagent
Thaum cov pa tshuaj nucleic acid kuaj pom kev thauj mus los hauv lub sijhawm ntev, (-20 ± 5) ℃ txias cia cov khoom thauj thiab thauj mus los yuav tsum xyuas kom lub cev nquag ntawm enzyme nyob rau hauv reagents tseem nyob. Txhawm rau kom paub tseeb tias qhov kub tau mus txog tus qauv, ob peb kilo ntawm dej khov qhuav xav tau rau txhua lub thawv ntawm nucleic acid kuaj reagent txawm tias tsawg dua 50g, tab sis nws tuaj yeem tsuas kav ntev li ob lossis peb hnub. Ntawm qhov pom ntawm kev coj ua kev lag luam, qhov nyhav nyhav tiag tiag ntawm cov reagents tawm los ntawm cov tuam ntxhab yog tsawg dua 10% (lossis deb dua tus nqi no) ntawm lub ntim. Feem ntau ntawm qhov hnyav los ntawm cov dej khov qhuav, cov ntawv ntim dej khov thiab cov thawv npaus, yog li tus nqi tsheb thauj khoom tau siab heev.

Thaum Lub Peb Hlis 2020, COVID-19 tau pib tawg tawm hauv qhov loj dua li txawv teb chaws, thiab qhov kev thov rau Novel Coronavirus nucleic acid kuaj pom tau muaj tseeb ntau zuj zus. Txawm hais tias tus nqi siab ntawm kev xa khoom reagents hauv cov saw txias, feem ntau cov tuam ntxhab tseem tuaj yeem lees txais nws vim tias qhov loj ntau thiab cov nyiaj tau los siab.

Txawm li cas los xij, nrog kev txhim kho ntawm lub teb chaws xa tawm cov cai rau kev tiv thaiv kev kis kab mob, nrog rau kev txhim kho cov kev tswj hwm hauv lub teb chaws ntawm cov neeg ntws thiab kev thauj mus los, muaj kev txuas ntxiv thiab tsis paub tseeb nyob rau hauv lub sijhawm thauj khoom ntawm kev rov ua dua, uas ua rau muaj teeb meem cov khoom lag luam zoo los ntawm kev thauj mus los. Ncua kev thauj mus los (lub sijhawm kev thauj mus los ntawm ib nrab ntawm ib hlis yog ib qho muaj heev) ua rau cov khoom lag luam tsis tu ncua thaum cov khoom mus txog tus neeg siv khoom. Qhov no tau teeb meem feem ntau nucleic acid reagents export lag luam.

Lyophilized thev naus laus zis rau PCR reagent tau pab thauj mus los ntawm Novel Coronavirus nucleic acid kuaj cov khoom thoob ntiaj teb

Cov lyophilized PCR reagents tuaj yeem thauj thiab khaws cia hauv chav sov, uas tuaj yeem tsis tsuas yog txo nqi kev thauj mus los, tab sis kuj zam dhau qhov teeb meem tsis zoo los ntawm kev thauj mus los. Yog li, lyophilizing reagent yog qhov zoo tshaj plaws los daws qhov teeb meem ntawm kev thauj khoom thauj.

Lyophilization cuam tshuam nrog khov ib qho kev daws teeb meem rau hauv lub xeev khov, thiab tom qab ntawd sublimate thiab cais cov dej vapor hauv qab qhov nqus. Lub tshuab ziab qhuav tseem nyob hauv cov thawv uas muaj tib qho kev ua ub ua no thiab kev ua. Muab piv nrog cov kua roj ua kom yooj yim, qhov muaj txhij-thev-zoo tshaj ntawm Kev Siv Tshuaj Neon Tus Kab Mob Coronavirus nucleic acid kuaj pom tau los ntawm cov Liming Bio muaj cov yam ntxwv li hauv qab no:

Tsis tshua muaj neeg muaj zog stability tshav kub: nws tuaj yeem nrog kev kho mob sawv ntsug ntawm 56 ℃ rau 60 hnub, thiab morphology thiab kev ua haujlwm ntawm reagent tsis hloov.
Qhov kub thiab txias cia khoom thiab tsheb thauj mus los: tsis tas yuav muab cov saw txias, tsis tas yuav khaws cia ntawm qhov kub qis ua ntej siv khoom, tso tawm qhov chaw txias txias.
Npaj-siv-siv: lyophilizing ntawm txhua yam, tsis xav tau kev teeb tsa system, zam qhov ploj ntawm cov khoom siv nrog viscosity siab xws li enzyme.
Multiplex cov hom phiaj hauv ib lub raj: lub hom phiaj nrhiav kev pab cuam tiv thaiv tus kab mob tshiab Coronavirus ORF1ab gene, N gene, S kom tsis txhob muaj tus kab mob genovariation. Txhawm rau kom txo cov cuav tsis zoo, tib neeg RNase P gene siv rau kev tswj hwm sab hauv, thiaj li ua tau raws li cov chaw kho mob xav tau coj mus kuaj kom zoo.


  • Yav dhau los:
  • Tom ntej:

  • Sau koj cov lus ntawm no thiab xa rau peb