Product Detail
Lub Koom Haum Saib Xyuas Kev Noj Qab Haus Huv Ntiaj Teb (WHO) qhia tias kev kuaj tshuaj tiv thaiv kab mob sai sai (Ag-RDRs) tuaj yeem muab txoj hauv kev sai thiab pheej yig dua los kuaj xyuas tus kab mob SARS-CoV-2 nquag tshaj li nucleic acid amplification tests (NAATs), thiab WHO kuj pom zoo. tias Ag-RDTs ua tau raws li qhov yuav tsum tau ua yam tsawg kawg nkaus tuaj yeem siv rau kev tshawb pom thawj zaug, kev sib cuag nrog kev tshawb nrhiav, thaum tshawb nrhiav kev sib kis thiab saib xyuas cov xwm txheej ntawm cov kab mob tshwm sim hauv cov zej zog.
Nta
● Kev Paub:Peb lub hom phiaj kuaj pom hauv ib qho kev sim
● tau tshaj:Kev yoog rau cov cuab yeej siv nrog CY5, FAM, VIC / HEX raws.
● Kev ua tau zoo heev:Siab rhiab heev thiab tshwj xeeb, LOD = 200 daim / ml.
Technical Parameter
| Packing Specification | 50 kuaj / khoom siv, 100 kuaj / khoom siv |
| Lub Hom Phiaj | ORF1ab, N, E |
| Siv tau Sample | Sputum, Oropharyngeal Swab |
| Txwv Kev Tshawb Nrhiav | 200 daim / ml |
| Total Coincidence Rate | 99.55% |
| Tus nqi Ct (CV,%) | ≤ 5.0% |
| Positive Coincidence Rate | 99.12% |
| Negative Coincidence Rate | 100% |
| Cia Tej yam kev mob thiab hnub tas sij hawm | Khaws ntawm -20 ± 5 ℃, thiab siv tau ib ntus rau 12 lub hlis. |
| Internal Control | Yog lawm |
| Catalog Number | A7793YF-50T, A7793YF-100T |
| Daim ntawv pov thawj | CE |
| Cov qauv | Nasopharyngeal swab, Oropharyngeal swab, Alveolar lavage kua, qaub ncaug thiab hnoos qeev |
| Siv tau Instrument | ABI 7500, Roche Lub Teeb Cycler 480Ⅱ, Roche Cobas z 480, SLAN-96P Real-Time PCR System |
Cov txheej txheem xeem
1. Nucleic Acid Extraction
Kev ua haujlwm yuav tsum tau ua raws li phau ntawv qhia ntawm cov khoom siv rho tawm.
2. Kev Npaj Ua Haujlwm:
1) Tshem tawm cov reagent thiab thaw lub reagent tag.Hloov qhov sib tov thiab centrifuge tam sim ntawd.N xeem cov tshuaj tiv thaiv (N = tus naj npawb ntawm cov qauv yuav tsum tau kuaj + zoo tswj + tsis zoo tswj + 1) yog npaj rau cov tshuaj tiv thaiv systems, feem, raws li nram no.
| Vomponents | Volume rau 1 cov tshuaj tiv thaiv kab mob | Volume rau N cov tshuaj tiv thaiv kab mob |
| Nucleic acid amplification cov tshuaj tiv thaiv Mix (A7793YF) | 18 µL | 18 µL * N |
| Enzyme sib tov | 2 µL | 2 µL * N |
| Tag nrho ntim | 20 µL | 20 µL * N |
2) Kev faib tawm cov tshuaj tiv thaiv: Cov tshuaj tiv thaiv tau sib xyaw thiab centrifuged, thiab txhua lub raj tau muab tso rau hauv qhov nyiaj ntawm 20μL hauv PCR raj tsim nyog rau lub tshuab hluav taws xob fluorescence PCR.
3. Chaw thau khoom
5μL ntawm cov qauv rho tawm nucleic acid, zoo tswj nucleic acid thiab tswj tsis zoo nucleic acid ntxiv rau cov tshuaj tiv thaiv kab mob, thiab tag nrho cov tshuaj tiv thaiv ntim yog 25μL.Fasten lub hau npog thiab txav mus rau qhov chaw kuaj amplification tom qab ob peb feeb ntawm centrifugation.
4. PCR Amplification Assay
1) Muab PCR cov tshuaj tiv thaiv raj rau hauv fluorescent PCR amplification instrument rau amplification detection.
2) Cycle parameter teeb tsa:
| Program | Tus lej ntawm lub voj voog | Kub | Cov tshuaj tiv thaiv lub sijhawm | |
| 1 | 1 | 50 ℃ | 10 feeb | |
| 2 | 1 | 95 ℃ | 30 sec | |
| 3 | 45 | 95 ℃ | 5 sec | |
| 60 ℃ | 30 sec | Fluorescence Collection | ||
3) Kev ntsuas kev ntsuas:
Cov kev kuaj pom tau raug teeb tsa rau FAM, VIC, ROX thiab CY5, sib xws rau ORF1ab, N gene, thiab E Gene, RNase P kev tswj hwm sab hauv, feem."Quencher Dye" thiab "Passive Reference" yog teem rau "Tsis muaj" rau ABI 7500 ntsuas.Teem Cov Kev Tswj Zoo, Kev Tswj Tsis Zoo, thiab Qauv (Tsis Paub) nyob rau hauv qhov kev txiav txim uas cov qauv sib raug, thiab teeb tsa tus qauv npe hauv kab "Sample Name" kem.
Rau X-POCH16, kev ua haujlwm thiab kev pabcuam yog raws li hauv qab no:
1) Tom qab kuaj tus kheej tiav lawm, qhib lub hau thiab muab cov tshuaj tiv thaiv PCR rau hauv cov haujlwm uas tau teev tseg hauv qhov ntsuas.
2) Pib los ntawm kev xaiv "Exper."kev xaiv.Xaiv qhov "Tag nrho" kev xaiv los yog manually xaiv qhov tshuaj tiv thaiv cheeb tsam ntawm sab laug ntawm lub vijtsam.
3) Xaiv qhov "LOAD" kev xaiv;xaiv qhov kev xeem ntawv;nyem "DONE" thiab "RUN".Qhov kev zov me nyuam siv sijhawm 30min42s kom tiav.
Cov kev kuaj pom ntawm qhov kev pab cuam Default yog teem rau FAM, VIC, ROX thiab CY5, sib xws rau ORF1ab, N noob, thiab E Gene, RNase P kev tswj sab hauv, raws li.
Lub voj voog parameter ntawm Default program yog raws li hauv qab no:
| Program | Tus naj npawb ntawm | Kub | Cov tshuaj tiv thaiv lub sijhawm |
| 1 | 1 | 50 ℃ | 2 min |
| 2 | 1 | 95 ℃ | 30 sec |
| 3 | 41 | 95 ℃ | 2 sec |
| 60 ℃ | 13 sec | Fluorescence |
5. Qhov chaw pib
Raws li cov duab txheeb xyuas, kho qhov pib tus nqi, qhov kawg tus nqi ntawm lub hauv paus thiab tus nqi pib (Pib tus nqi thiab tus nqi kawg yog pom zoo kom muab tso rau 3 thiab 15 raws li, thiab qhov amplification nkhaus ntawm qhov kev tswj tsis zoo yog kho kom tiaj tus lossis qis dua qhov pib kab), nyem Kev Tshawb Fawb kom tau txais qhov kev tshuaj ntsuam tus qauv Ct tus nqi.Saib cov txiaj ntsig hauv Report interface.
6. Kev Tswj Xyuas Zoo
Txhua qhov kev tswj ntawm cov khoom siv yuav tsum ua raws li cov cai hauv qab no nrog 'S' nkhaus, txwv tsis pub qhov kev sim ua tsis raug.
| Tshawb nrhiav cov channel | Kev tswj tsis zoo | Kev tswj zoo |
| FAM(ORF1ab) | Tsis muaj Ct | Ct≤38 |
| VIC(N) | Tsis muaj Ct | Ct≤38 |
| ROX (E) | Tsis muaj Ct | Ct≤38 |
| CY5 (RP) | Tsis muaj Ct | Ct≤38 |
【Kev txiav tawm tus nqi】
Raws li cov txiaj ntsig ntawm 100 oropharyngeal swab kuaj thiab 100 cov hnoos qeev, thiab nrog ROC nkhaus txoj kev, qhov txiav tawm tus nqi ntawm OFR1ab, N genes E Gene ntawm cov khoom siv no yog Ct = 38
FAQ
Hauv cov khoom siv no, primers thiab probes ntawm real-time fluorescent PCR thev naus laus zis yog tsim los rau thaj chaw khaws cia thiab tshwj xeeb ntawm ORF1ab, N thiab E noob ntawm 2019-nCoV feem.Thaum lub sij hawm PCR amplification, qhov kev sojntsuam khi rau lub template, thiab 5'-kawg reporter pab pawg neeg sojntsuam yog cleaved los ntawm lub Taq enzyme (5'→ 3' exonuclease kev ua), yog li txav deb ntawm lub quenching pab pawg neeg los tsim ib tug fluorescent teeb liab. .Lub sijhawm ua kom pom tseeb ntawm lub sijhawm tiag tiag yog txiav txim siab raws li qhov teeb meem fluorescence pom, thiab cov qauv Ct tus nqi raug xam.FAM , VIC thiab ROX fluorophores tau sau rau ORF1ab gene, N gene thiab E noob sojntsuam.Los ntawm kev siv ib qho kev sim, kev kuaj pom zoo ntawm peb lub noob saum toj no ntawm 2019-nCoV tuaj yeem ua tiav ib txhij.
Cov khoom siv tau muab nrog kev tswj hwm sab hauv tsom rau RNase P gene los saib xyuas cov qauv kuaj mob, tuav, rho tawm thiab RT-PCR txheej txheem kom tsis txhob muaj cov txiaj ntsig tsis zoo.Kev tswj sab hauv yog sau nrog CY5 fluorescent pawg.
1. Txheeb xyuas thiab txhais cov txiaj ntsig kev xeem thaum lub ntsuas ntsuas yog qhov qub, thiab kev tswj xyuas qhov zoo, kev tswj tsis zoo thiab kev ntsuas kev ntsuas sab hauv ua tau raws li tus qauv tswj tau zoo.
2. Lub amplification nkhaus ntawm kev tswj sab hauv (CY5) qhia ib tug raug S nkhaus thiab Ct ≤ 38, kev txhais cov ntsiab lus ntawm lub hom phiaj noob tau raug raws li nram no.
| Tshawb nrhiav cov channel | Txhais cov txiaj ntsig ntawm cov hom phiaj | ||
| FMA | VIC (N gene) | ROX (E noob) | |
| Ct≤38 | Ct≤38 | Ct≤38 | Nrog rau tus qauv S amplification nkhaus, tus nqi Ct yog ≤38, cov hom phiaj sib txuas yog qhov zoo. |
| 38 Ct. 40 | 38 Ct. 40 | 38 Ct. 40 | Nrog ib tug raug S amplification nkhaus, rov ntsuam xyuas tus coj lub hom phiaj ntawm tus qauv dua. Yog hais tias tus nqi Ct 40 nrog ib tug raug S amplification nkhaus, tus coj lub hom phiaj yog zoo;Yog tias tus nqi Ct ≥40, cov hom phiaj sib txuas yog qhov tsis zoo |
| Txt≥40 | Txt≥40 | Txt≥40 | Cov hom phiaj sib thooj yog qhov tsis zoo |
Interpreteration of results for 2019-nCoV:
Raws li cov txiaj ntsig ntawm ORF1ab, N gene thiab E noob, kev txhais lus raws li hauv qab no:
1) Yog tias ob lossis peb lub noob ntawm cov noob kuaj pom muaj qhov zoo, 2019-nCoV yog qhov zoo;
2) Yog tias tsuas yog ib qho lossis tsis yog ib qho ntawm cov noob tau pom zoo, 2019-nCoV yog qhov tsis zoo.
Nco tseg: Qhov kev nthuav dav nkhaus ntawm cov qauv zoo yuav tsum yog nrog tus qauv S nkhaus.Txawm li cas los xij, yog tias lub hom phiaj concentration siab dhau lawm, kev tswj hwm tus qauv sab hauv yuav tsis nthuav dav thiab cov qauv tuaj yeem txiav txim ncaj qha raws li qhov zoo.Yog tias ib qho ntawm ob lub hom phiaj tau txais Ct≤38, 2019-nCoV yog qhov zoo.Yog tias ib qho ntawm ob lub hom phiaj tau txais Ct≥40, 2019-nCoV yog qhov tsis zoo.Yog tias Ct≥40, lossis tsis muaj nqi, kev txhais cov txiaj ntsig ntawm lub hom phiaj yog qhov tsis zoo.
3. Yog tias tag nrho cov Ct qhov tseem ceeb ntawm FAM, VIC, ROX thiab Cy5 raws yog ntau tshaj 38 lossis tsis muaj qhov pom tseeb ntawm S amplification nkhaus:
1) Muaj / muaj cov khoom (s) hauv cov qauv uas inhibit PCR cov tshuaj tiv thaiv.Nws raug nquahu kom dilute tus qauv kom rov kuaj dua.
2) Cov txheej txheem ntawm nucleic acid extraction yog txawv txav, yog li nws tau pom zoo kom rov rho tawm
nucleic acid rau rov kuaj dua.
3) Cov qauv no tsis yog ib qho qauv tsim nyog thaum lub sijhawm ua piv txwv, lossis degraded
thaum thauj thiab khaws cia.
Lawv yog 2 txoj hauv kev uas peb tuaj yeem kuaj pom qhov xwm txheej no: NAAT thiab Antigen.
(Los ntawm CDC ntawm Los Angeles)
