Novel Coronavirus (SARS-CoV-2) Multiplex Real-Time PCR Pusa
Lenei sili atu maaleale, sauni-e-faʻaaoga pusa PCR o loʻo avanoa i le lyophilized faʻataʻitaʻiga (liu aisa-faʻamago gaioiga) mo se taimi umi teuina. O le pusa e mafai ona aveina ma teu i le potu vevela ma e mausali mo le tasi le tausaga. Taʻitasi faagaau o premix aofia uma reagents manaʻomia mo le PCR amplification, aofia ai Reverse-transcriptase, Taq polymerase, primers, probes, ma dNTPs substrates. E naʻo le faʻaopopoina o le 13ul distill vai ma le 5ul aveʻese mai le RNA template, ona mafai lea ona tamoʻe ma faʻateleina i mea fai a le PCR.
O le masini qPCR tatau ona faʻamalieina manaʻoga nei:
1. Fetaui 8 fasimea PCR paipa voluma 0.2 ml
2. Ia sili atu ma le fa auala vaʻaia.
Channel |
Fiafia (nm) |
Emission (nm) |
Lanu Muai Fofoina |
1. |
470 |
525 |
FAM, SYBR Green I |
2 |
523 |
564 |
VIC, HEX, TET, JOE |
3. |
571 |
621 |
ROX, TEXAS-Mumu |
4 |
630 |
670 |
CY5 |
PCR-fausaga opea:
7500Real-Time PCR System, Biorad CF96, iCycler iQ ™ Real-Time PCR Detection System, Stratagene Mx3000P, Mx3005P
Le faigata o felauaiga malulu filifili o Novel Coronavirus nucleic acid mauaina mauaina
Pe a masani ona feaveaʻia vaʻaia vaʻavaʻaia nucleic acid i mea mamao, o le (-20 ± 5) ℃ malulu filifili filifili ma felauaʻiga e manaʻomia e faʻamautinoa ai o le bioactive o le enzyme i le reagents tumau malosi. Ina ia mautinoa o le vevela oʻo i le tulaga masani, e tele kilokalama o le aisa mago e manaʻomia mo atigipusa taʻitasi o le nucleic acid suʻega reagent tusa o lalo ifo o le 50g, ae e naʻo le lua pe tolu aso e mafai ai. I luga o le vaʻaiga a le aufaipisinisi faʻataʻitaʻi, o le mamafa moni o reagents na tuʻuina mai e le au gaosi e laititi atu i le 10% (pe sili mamao atu i lo lenei tau) o le container. Ole tele ole mamafa e sau mai le aisa mago, aisa ma pusa foam, o lona uiga e taugata tele le tau o femalagaiga.
I le masina o Mati 2020, na amata ai ona malepe le COVID-19 i se tulaga tele i fafo atu o le atunuʻu, ma o le manaʻoga o le Novel Coronavirus nucleic acid detagent ua faʻateleina tele. E ui lava i le taugata o le auina atu i fafo o le reagents i le malulu filifili, tele o tagata gaosi mafai lava ona taliaina ona o le tele tele ma maualuga tupe mama.
Peitaʻi, i le faʻaleleia atili o tulafono faʻavaomalo mo oloa auina atu i fafo, faʻapea foi ma le faʻaleleia o le pulega a le atunuʻu i luga o le tafe o tagata ma logistics, o loʻo i ai le faʻaopoopoga ma le le mautonu i le taimi o femalagaaiga a aliʻi, na mafua ai faʻafitauli iloga o oloa. e ala i femalagaiga. O le umi o femalagaaiga taimi (femalagaaiga taimi pe a ma le afa masina e matua masani ai) e tau atu ai i le tele o taimi oloa le manuia pe a oʻo le oloa i le tagata o tausia. Lenei ua faʻalavelaveina le tele o nucleic acid faʻalauteleina pisinisi auina atu i fafo.
Lyophilized tekonolosi mo PCR reagent fesoasoani i le felauaiga o Novel Coronavirus nucleic acid mauaina mauaina reagent lalolagi atoa
O le lyophilized PCR reagents mafai ona feaveaʻi ma teuina i le potu vevela, lea e le gata mafai faʻaititia ai le tau o felauaiga, ae faʻapea foi ona aloese ai mai le lelei faʻafitauli mafua mai i auala felauaiga. Ole mea lea, ole faʻamalosi tino ole auala ole auala pito sili lea e fofo ai le faʻafitauli ole felauaiga i fafo.
Lyophilization aofia ai le faʻamaʻaina o se fofo i totonu o se mautu tulaga, ona faʻamamaina ma tuueseese le ausa vai i lalo o lagona gaogao. O le solute solute o loʻo tumau pea i totonu o le koneteina ma le tutusa tuʻufaʻatasiga ma gaioiga. Faʻatusatusa i vailaʻau masani suavai, o le atoa-vaega lyophilized Novel Coronavirus nucleic acid mauaina vailaʻau na faia e Liming Bio ei ai uiga nei:
Matua mausali malosi faʻamautuina: e mafai ma le tu togafitiga i le 56 ℃ mo 60 aso, ma le morphology ma faʻatinoina o le reagent tumau le suia.
Teuina masani o le vevela ma felauaiga: leai se manaʻoga mo malulu filifili, leai se mea e teu ai i le maualalo vevela i luma o le faʻamaufaʻailogaina, faʻamatuʻu atoa le malulu teuina avanoa.
Sauni-e-faʻaaoga: lyophilizing o vaega uma, leai se manaʻoga mo le faʻatulagaina o le tino, aloese mai le leiloa o vaega ma maualuga viscosity pei o le enzyme.
Multiplex sini i le tasi faagaau: o le faʻamoemoe mauaina e aofia ai le coronavirus ORF1ab gene, N gene, S gene e aloese ai mai le siama genovariation. Ina ia mafai ona faʻaititia le sese le lelei, o le tagata RNase P gene e faʻaaogaina e avea ma faʻatonutonu i totonu, ina ia mafai ai ona ausia le falemaʻi manaʻoga mo faʻataʻitaʻiga lelei lelei.