Novel Coronavirus (SARS-CoV-2) Multiplex Real-Time PCR Kit
Zida za PCR zomverera kwambiri, zokonzeka kugwiritsa ntchito zimapezeka mumtundu wa lyophilized (njira yowumitsa zowuma) kuti zisungidwe kwa nthawi yayitali.Chombocho chikhoza kunyamulidwa ndikusungidwa kutentha kwa chipinda ndipo chimakhala chokhazikika kwa chaka chimodzi.Chubu chilichonse cha premix chimakhala ndi ma reagents ofunikira pakukulitsa kwa PCR, kuphatikiza Reverse-transcriptase, Taq polymerase, primers, probes, ndi dNTPs substrates.Zimangofunika kuwonjezera madzi osungunuka a 13ul ndi template ya 5ul yotulutsidwa ya RNA, ndiye kuti imatha kuthamanga ndikukulitsidwa pazida za PCR.
Makina a qPCR akuyenera kukwaniritsa izi:
1. Fit 8 strip PCR chubu voliyumu 0,2 ml
2. Khalani ndi njira zopitilira zinayi zozindikirira:
| Channel | Chisangalalo (nm) | Kutulutsa (nm) | Madyerero Osasinthika |
| 1. | 470 | 525 | FAM, SYBR Green I |
| 2 | 523 | 564 | VIC, HEX, TET, JOE |
| 3. | 571 | 621 | ROX, TEXAS-RED |
| 4 | 630 | 670 | CY5 |
PCR-nsanja:
7500Real-Time PCR System, Biorad CF96, iCycler iQ™ Real-Time PCR Detection System, Stratagene Mx3000P, Mx3005P
Kuvuta kwamayendedwe ozizira a Novel Coronavirus nucleic acid discovery reagent
Pamene ma reagents odziwika bwino a nucleic acid amanyamulidwa patali, (-20 ± 5) ℃ kusungirako kuzizira kwa unyolo ndi zoyendera zimafunika kuwonetsetsa kuti ma enzyme mu reagents amakhalabe achangu.Kuonetsetsa kuti kutentha kumafika pamlingo, ma kilogalamu angapo a ayezi wouma amafunikira pabokosi lililonse la nucleic acid kuyesa reagent ngakhale zosakwana 50g, koma zimatha masiku awiri kapena atatu okha.Potengera machitidwe amakampani, kulemera kwenikweni kwa ma reagents operekedwa ndi opanga ndi ochepera 10% (kapena ocheperako kuposa mtengo uwu) wa chidebecho.Zolemera zambiri zimachokera ku ayezi wouma, mapaketi a ayezi ndi mabokosi a thovu, kotero kuti mtengo wamayendedwe ndi wokwera kwambiri.
Mu Marichi 2020, COVID-19 idayamba kufalikira kumayiko ena, ndipo kufunikira kwa Novel Coronavirus nucleic acid kuzindikira reagent kudakula kwambiri.Ngakhale mtengo wamtengo wapatali wotumizira ma reagents mu unyolo wozizira, opanga ambiri amatha kuvomerezabe chifukwa cha kuchuluka kwakukulu komanso phindu lalikulu.
Komabe, ndi kusintha kwa ndondomeko za dziko zogulitsa katundu zotsutsana ndi mliri, komanso kukweza ulamuliro wa dziko pa kayendetsedwe ka anthu ndi kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka kayendetsedwe ka anthu. ndi mayendedwe.Nthawi yowonjezereka yoyendera (nthawi yoyendera pafupifupi theka la mwezi ndiyofala kwambiri) imayambitsa kulephera kwazinthu pafupipafupi pomwe mankhwalawo afika kwa kasitomala.Izi zasokoneza mabizinesi ambiri ogulitsa ma nucleic acid.
Tekinoloje ya Lyophilized ya PCR reagent idathandizira mayendedwe a Novel Coronavirus nucleic acid kuzindikira reagent padziko lonse lapansi.
The lyophilized PCR reagents akhoza kunyamulidwa ndi kusungidwa kutentha kwa firiji, zomwe sizingangochepetsa mtengo wa zoyendetsa, komanso kupewa mavuto omwe amadza chifukwa cha kayendetsedwe ka kayendetsedwe kake.Choncho, lyophilizing ndi reagent ndiyo njira yabwino yothetsera vuto la kutumiza kunja.
Lyophilization imaphatikizapo kuziziritsa njira kukhala yolimba, ndiyeno sublimate ndikulekanitsa nthunzi wamadzi pansi pa vacuum.The solute zouma amakhala mu chidebe ndi zikuchokera chimodzimodzi ndi ntchito.Poyerekeza ndi ma reagents ochiritsira amadzimadzi, gawo lonse la lyophilized Novel Coronavirus nucleic acid kuzindikira reagent yopangidwa ndi Liming Bio ili ndi izi:
Kukhazikika kwa kutentha kwambiri:imatha ndi chithandizo choyima pa 56 ℃ kwa masiku 60, ndipo mawonekedwe ndi magwiridwe antchito a reagent amakhalabe osasinthika.
Kusungirako kutentha kwanthawi zonse ndi zoyendera:palibe chifukwa cha unyolo wozizira, palibe chifukwa chosungira kutentha kochepa musanamasule, kumasula kwathunthu malo osungira ozizira.
Zakonzeka kugwiritsidwa ntchito:lyophilizing onse zigawo zikuluzikulu, palibe chifukwa dongosolo kasinthidwe, kupewa imfa ya zigawo zikuluzikulu ndi mkulu mamasukidwe akayendedwe monga enzyme.
Multiplex targets mu chubu chimodzi:chandamale chodziwikiratu chimakhudza jini ya coronavirus ya ORF1ab, jini ya N, jini ya S kupewa kufalikira kwa kachilomboka.Pofuna kuchepetsa zolakwika zabodza, jini ya RNase P yaumunthu imagwiritsidwa ntchito ngati chiwongolero chamkati, kuti akwaniritse zosowa zachipatala zowongolera khalidwe lachitsanzo.
