New Coronavirus(SARS-Cov-2) Nucleic Acid Detection Kit

New Coronavirus(SARS-Cov-2) Nucleic Acid Detection Kit

(Fluorescent RT-PCR Probe Method) Product Manual

【Produc uat name 】New Coronavirus(SARS-Cov-2) Nucleic Acid Detection Kit(Fluorescent RT-PCR Probe Method)

【Packaging specua ifications 】25 Tests/Kit

【Intended ushnub nyoog】

Cov khoom siv no yog siv rau kev kuaj pom zoo ntawm nucleic acid los ntawm tus kabmob tshiab no hauv nasopharyngeal swabs, oropharyngeal (xab caj pas) swabs, anterior nasal swabs, mid-turbinate swabs, qhov ntswg ntxuav thiab qhov ntswg aspirates los ntawm cov tib neeg uas xav tias yog COVID19 los ntawm lawv tus kws kho mob. Kev kuaj pom ntawm ORF1ab thiab N noob ntawm tus kabmob tshiab tuaj yeem siv rau kev kuaj mob pabcuam thiab kev saib xyuas kabmob kis kabmob tshiab.

【Principles of the procedure 】

Cov khoom siv no yog tsim los rau cov kev sojntsuam TaqMan tshwj xeeb thiab cov primers tshwj xeeb tsim los rau tus kabmob tshiab (SARS-Cov-2) ORF1ab thiab N gene sequences. Cov tshuaj tiv thaiv PCR muaj 3 pawg ntawm cov primers tshwj xeeb thiab cov tshuaj fluorescent rau kev tshawb pom cov hom phiaj, thiab cov txheej txheem ntxiv ntawm cov primers thiab fluorescent probes yog siv los ua tus txheej txheem sab hauv ntawm cov khoom siv los kuaj xyuas cov noob hauv tsev endogenous.

Lub hauv paus ntsiab lus ntawm kev xeem yog tias qhov tshwj xeeb fluorescent sojntsuam yog zom thiab degraded los ntawm exonuclease kev ua ntawm Taq enzyme nyob rau hauv cov tshuaj tiv thaiv PCR, thiaj li hais tias cov reporter fluorescent pawg thiab quenched fluorescent pawg yog sib cais, kom lub fluorescence xyuas system tau txais ib tug fluorescent. teeb liab, thiab tom qab ntawd Los ntawm cov txiaj ntsig zoo ntawm PCR amplification, lub teeb liab fluorescence ntawm qhov kev sojntsuam ncav cuag tus nqi pib-Ct tus nqi (Cycle threshold). Nyob rau hauv rooj plaub uas tsis muaj lub hom phiaj amplicon, pawg neeg sau xov xwm ntawm qhov kev sojntsuam yog nyob ze rau pawg quenching. Lub sijhawm no, fluorescence resonance zog hloov pauv tshwm sim, thiab fluorescence ntawm pawg neeg sau xov xwm raug tua los ntawm pab pawg quenching, kom cov teeb liab fluorescent tsis tuaj yeem kuaj pom los ntawm fluorescent PCR ntsuas.

Txhawm rau saib xyuas kev siv cov tshuaj reagents thaum lub sijhawm kuaj, cov khoom siv tau nruab nrog kev tswj xyuas qhov zoo thiab tsis zoo: qhov kev tswj xyuas zoo muaj lub hom phiaj recombinant plasmid, thiab kev tswj tsis zoo yog dej Distilled, uas yog siv los saib xyuas ib puag ncig cov pa phem. Nws raug nquahu kom teeb tsa kev tswj hwm qhov zoo thiab kev tswj tsis zoo ib txhij thaum kuaj.

【Main components 】

Nco tseg:(1) Cov khoom sib txawv hauv cov khoom siv sib txawv tsis tuaj yeem sib xyaw lossis sib hloov.

(2) Npaj koj tus kheej reagent: Nucleic acid extraction kit.

【Storage condnwsons thiab expiration date 】

For BST-SARS-25:Kev thauj mus los thiab khaws cia ntawm -20 ± 5 ℃ rau lub sijhawm ntev.

For BST-SARS-DR-25:Kev thauj mus los ntawm chav tsev kub. Khaws ntawm -20 ± 5 ℃ rau lub sijhawm ntev.

Tsis txhob rov qab khov-thaw cycles. Lub sijhawm siv tau yog teem sijhawm rau 12 lub hlis.

Saib daim ntawv lo rau hnub tsim thiab siv.

Tom qab qhib thawj zaug, cov reagent tuaj yeem khaws cia ntawm -20 ± 5 ° C tsis pub dhau 1 lub hlis lossis txog rau thaum kawg ntawm lub sijhawm reagent, seb hnub twg los ua ntej, kom tsis txhob rov qab khov-thaw mus, thiab tus naj npawb ntawm cov reagent khov. -thaw cycles yuav tsum tsis pub tshaj 6 zaug.

【Applicable instrument】ABI 7500, SLAN-96P, Roche-LightCycler-480.

【Sample requirements 】

1.Ua piv txwv hom: Extracted nucleic acid tov.

2.Sample cia thiab tsheb thauj mus los: Khaws ntawm-20 ± 5 ℃ rau 6 lub hlis.Freeze thiab thaw cov qauv tsis pub ntau tshaj 6 zaug.

【Tyoging method】

1.Nucleic acid extraction

Xaiv cov khoom siv tsim nyog nucleic acid rho tawm cov kab mob nucleic acid, thiab ua raws li cov lus qhia siv cov khoom siv sib xws. Nws raug pom zoo kom siv Nucleic acid extraction thiab purification cov khoom tsim los ntawm Yixin Bio-Tech (Guangzhou) Co., Ltd. los yog sib npaug nucleic acid purification cov khoom.

2.  Rua eaction rovnt prepua aration

2.1 For BST-SARS-25:

(1) Tshem tawm SARS-Cov-2 Mix thiab Enzyme Mix, tag nrho yaj ntawm chav tsev kub sib tov kom huv si los ntawm Vortex ntaus ntawv thiab tom qab ntawd centrifuge luv luv.

(2) 16.5uL Enzyme Mix tau ntxiv rau 358.5uL SARS-Cov-2 Mix thiab sib xyaw kom zoo kom tau txais cov tshuaj sib xyaw ua ke.

(3) Npaj ib qho huv si 0.2 mL PCR octal raj thiab kos nws nrog 15uL ntawm cov tshuaj sib xyaw saum toj no rau ib qhov dej.

(4) Ntxiv 15 μL ntawm purified nucleic acid tov, kev tswj zoo thiab kev tswj tsis zoo, thiab ua tib zoo npog lub octal raj cap.

(5) Sib tov zoo los ntawm inverting upside down, thiab sai centrifuge kom concentrate cov kua nyob rau hauv qab ntawm lub raj.

1