Novel Coronavirus (SARS-CoV-2) Multiplex Real-Time PCR Kit
Sesebelisoa sena sa PCR se hlokolosi haholo se ikemiselitseng ho sebelisoa se fumaneha ka sebopeho sa lyophilized (ts'ebetso ea ho omisa leqhoa) bakeng sa polokelo ea nako e telele. Thepa e ka tsamaisoa ebe e bolokoa ka mocheso oa kamore 'me e tsitsitse selemo se le seng. Phala e 'ngoe le e' ngoe ea premix e na le li-reagents tsohle tse hlokahalang molemong oa ntlafatso ea PCR, ho kenyeletsoa Reverse-transcriptase, Taq polymerase, li-primers, probes le li-subNate tsa dNTPs. E hloka feela ho eketsa metsi a 13ul a khabisitsoeng le template ea RNA e ntšitsoeng ka 5ul, ebe e ka tsamaisoa le ho holisoa ho lisebelisoa tsa PCR.
Mochini oa qPCR o lokela ho fihlela litlhokahalo tse latelang:
1. Fitisa 8 strip tube ea PCR bophahamo ba 0.2 ml
2. Ho ba le metjha e fetang e mene ea phumano:
Mocha |
Nyakallo (nm) |
Mohlapo (nm) |
Li-dae tsa pele ho nako |
1. |
470 |
525 |
FAM, SYBR Green ke |
2 |
523 |
564 |
VIC, HEX, TET, JOE |
3. |
571 |
621 |
ROX, TEXAS-Khubelu |
4 |
630 |
670 |
EA-CY5 |
Lipolanete tsa PCR:
7500Real-Time PCR System, Biorad CF96, iCycler iQ ™ Real-Time PCR Detection System, Stratagene Mx3000P, Mx3005P
Bothata ba lipalangoang tsa ketane e batang ea Reagent ea Novel Coronavirus nucleic acid
Ha li-reagents tse tloaelehileng tsa ho tseba li-acid li tsamaisoa sebakeng se selelele, (-20 ± 5) (poloko ea ketane e batang le lipalangoang lia hlokahala ho netefatsa hore bioactive ea enzyme ho li-reagents e lula e sebetsa. Ho etsa bonnete ba hore mocheso o fihla maemong, ho hlokahala li-kilogram tse 'maloa tsa leqhoa le ommeng bakeng sa lebokose le leng le le leng la tlhahlobo ea acid ea nucleic le ka tlase ho 50g, empa e ka nka matsatsi a mabeli kapa a mararo feela. Ho latela ts'ebetso ea indasteri, boima ba 'nete ba li-reagents bo fanoeng ke baetsi bo ka tlase ho 10% (kapa bo tlase haholo ho boleng bona) ba setshelo. Boima bo bongata bo tsoa leqhoeng le ommeng, lipaketeng tsa leqhoa le mabokoseng a foam, kahoo litšenyehelo tsa lipalangoang li phahame haholo.
Ka Hlakubele 2020, COVID-19 e qalile ho hlaha haholo kantle ho naha, mme tlhoko ea Novel Coronavirus nucleic acid detection reagent e eketsehile haholo. Leha ho na le litšenyehelo tse phahameng tsa ho rekisa li-reagents ka har'a ketane e batang, baetsi ba bangata ba ntse ba ka e amohela ka lebaka la bongata le phaello e phahameng.
Leha ho le joalo, ka ntlafatso ea maano a naha a kantle ho naha bakeng sa lihlahisoa tse khahlano le seoa, hammoho le ntlafatso ea taolo ea naha mabapi le phallo ea batho le lisebelisuoa, ho na le katoloso le ho se ts'oanehe nakong ea lipalangoang tsa li-reagents, tse bakileng mathata a hlaheletseng a sehlahisoa a bakileng ka lipalangoang. Nako ea lipalangoang e atolositsoeng (nako ea lipalangoang e ka bang halofo ea khoeli e atile haholo) e lebisa khafetsa sehlahisoa ha sehlahisoa se fihla ho moreki Sena se tšoentse likhoebo tse ngata tsa thekiso ea kantle ho naha tsa acid ea nucleic.
Theknoloji ea Lyophilized bakeng sa reagent ea PCR e thusitse ho tsamaisoa ha Novel Coronavirus nucleic acid detection reagent lefats'eng ka bophara
Li-reagents tsa PCR tse nang le lyophilized li ka tsamaisoa le ho bolokoa ka mocheso oa kamore, tse ke keng tsa fokotsa litšenyehelo tsa lipalangoang feela, empa hape li qobe mathata a boleng a bakoang ke ts'ebetso ea lipalangoang. Ka hona, lyophilizing reagent ke tsela e molemohali ea ho rarolla bothata ba lipalangoang tsa kantle ho naha.
Lyophilization e kenyelletsa ho qhibiliha tharollo hore e be maemong a tiileng, ebe o beha le ho arola mouoane oa metsi tlasa boemo ba leqhubu. Motsoako o omisitsoeng o lula ka har'a setshelo ka sebopeho le ts'ebetso e ts'oanang. Ha e bapisoa le li-reagents tse tloaelehileng tsa metsi, karolo e felletseng ea lyophilized Novel Coronavirus nucleic acid detection reagent e hlahisitsoeng ke Liming Bio e na le litšobotsi tse latelang:
Mofuthu o matla oa mocheso: e ka ts'oaroa kalafo ho 56 ℃ matsatsi a 60, mme morphology le ts'ebetso ea reagent li lula li sa fetohe.
Polokelo e tloaelehileng ea mocheso le lipalangoang: Ha ho na tlhoko ea ketane e batang, ha ho na lebaka la ho boloka mocheso o tlase pele o koaloa, o lokolle ka botlalo sebaka sa polokelo e batang.
Itokiselitse ho sebelisa: lyophilizing ea likarolo tsohle, ha ho na tlhoko ea ho hlophisa sistimi, ho qoba ho lahleheloa ke likarolo tse nang le viscosity e phahameng joalo ka enzyme
Lipheo tsa Multiplex ka har'a tube e le 'ngoe: sepheo sa ho lemoha se akaretsa mofuta oa ponahalo ea coronavirus ORF1ab, mofuta oa N, S lefutso ho qoba kokoana-hloko ea genovariation. Bakeng sa ho fokotsa liphoso tse fosahetseng, lefutso la motho la RNase P le sebelisoa joalo ka taolo ea kahare, ho fihlela tlhoko ea bongaka ea sampole ea taolo ea boleng.