IKhiti entsha yokuFumana iNucleic Acid yeNucleic (SARS-Cov-2).
New Coronavirus(SARS-Cov-2) Nucleic Acid Detection Kit
(Fluintsimbiscent RT-PCR Probe Method) Product Manual
【Product name 】IKhithi yokuFumana iNucleic Acid eNtsha (SARS-Cov-2)(iNdlela yokuHlola iFluorescent RT-PCR)
【Packaging specukuba ndications 】Iimvavanyo ezingama-25/Ikhithi
【Intended usubudala】
Le kiti isetyenziselwa ukuchongwa okusemgangathweni kwe-nucleic acid kwi-coronavirus entsha kwi-nasopharyngeal swabs, i-oropharyngeal (umqala) swabs, i-anterior nasal swabs, i-mid-turbinate swabs, ukuhlamba ngeempumlo kunye ne-aspirates yeempumlo ezivela kubantu abarhanelwa ukuba bane-COVID19 ngumboneleli wabo wezempilo. Ukufunyaniswa kwe-ORF1ab kunye nofuzo lwe-N ye-coronavirus entsha inokusetyenziselwa uxilongo oluncedisayo kunye nokujongwa kwesifo sosulelo olutsha lwe-coronavirus.
【Principles of the procedure 】
Le khithi yenzelwe i-TaqMan probes ezithile kunye neeprimers ezithile eziyilelwe inoveli coronavirus (SARS-Cov-2) ORF1ab kunye nolandelelwano lwemfuza ye-N. Isisombululo se-PCR reaction iqulathe iiseti ezi-3 zeeprimers ezithile kunye ne-fluorescent probes zokubona okujoliswe kuko, kunye neseti eyongezelelweyo yeeprimers ezithile kunye ne-fluorescent probes isetyenziswa njengomgangatho wangaphakathi wolawulo lwekhithi yokukhangela i-endogenous housekeeping genes.
Umgaqo wovavanyo kukuba i-probe ethile ye-fluorescent igaywe kwaye ithotywe ngumsebenzi we-exonuclease we-enzyme ye-Taq kwi-reaction ye-PCR, ukuze iqela lentatheli ye-fluorescent kunye neqela elicinyiweyo le-fluorescent lahlulwe, ukuze inkqubo yokubeka iliso ye-fluorescence inokufumana i-fluorescent. uphawu, kwaye emva koko Ngokusebenzisa isiphumo sokutyebisa kwe-PCR yokukhulisa, isignali ye-fluorescence ye-probe ifikelela kwi-set threshold value-Ct value (i-Cycle threshold). Kwimeko yokungabikho kwe-amplicon ekujoliswe kuyo, iqela lentatheli ye-probe lisondele kwiqela lokucima. Ngeli xesha, i-fluorescence resonance energy transfer iyenzeka, kwaye i-fluorescence yeqela lentatheli icinyiwe liqela lokucima, ukuze isignali ye-fluorescent ingabonakali ngesixhobo se-PCR se-fluorescent.
Ukuze kubekwe esweni ukusetyenziswa kwe-reagents ngexesha lovavanyo, ikiti ixhotywe ngolawulo oluhle kunye nolubi: ulawulo oluhle luqulethe indawo ekujoliswe kuyo i-plasmid edibeneyo, kunye nolawulo olubi luyi-Distilled water, esetyenziselwa ukubeka iliso ukungcoliseka kwendalo. Kunconywa ukuseta ulawulo olulungileyo kunye nolawulo olubi ngaxeshanye xa kuhlolwa.
【Main comiponits 】
| Cat. No. | BST-SARS-25 | BST-SARS-DR-25 | Ikhomponiinto | |
| Name | Specuqinisekiso | Quantinto | Quantinto | |
| Ulawulo oluhle | 180 μL/ibhotyana | 1 | 1 | Iiplasmids ezenziwe ngokwenziwa, Amanzi adityanisiweyo |
| Ulawulo olubi | 180 μL/ibhotyana | 1 | 1 | Mamanzi amisiwe |
| SARS-Cov-2 Mix | 358.5 μL/ivial | 1 | / | Izibini zeprimer ezikhethekileyo, iiprobes ezithile zokubhaqwa kwefluorescent, dNTPs, , MgCl2, KCl, Tris-Hcl, Distilled water, njl. |
| Umxube we-Enzyme | 16.5 μL/ivial | 1 | / | I-Taq enzymes, i-reverse transcriptase, i-UNG enzymes, njl. |
| SARS-Cov-2 Mix(Lyophilised) | Iimvavanyo ze-25 / i-vial | / | 1 | Izibini ezikhethekileyo zeprimer, i-probes ethile yokubona i-fluorescent probes, i-dNTPs, i-Taq enzymes, i-reverse transcriptase, i-Distilled water, njl. |
| 2x Isithinteli | 375 μL/ibhotyana | / | 1 | MgCl2, KCl, Tris-Hcl, Distilled water, njl. |
Phawula:( 1) Amacandelo kwiikiti zeebhetshi ezahlukeneyo azikwazi ukuxutywa okanye ukutshintshwa.
(2) Lungiselela i-reagent yakho: Ikhithi yokukhupha i-Nucleic acid.
【Storage condinto mnaons kwaye expiration date 】
For BST-SARS-25:Ezothutho kunye nevenkile kwi -20±5℃ ixesha elide.
For BST-SARS-DR-25:Ukuthutha kwiqondo lokushisa. Gcina kwi -20±5℃ ixesha elide.
Kuphephe ukuphindaphinda imijikelo yokunyibilika komkhenkce. Ixesha lokuqinisekisa limiselwe iinyanga ezili-12.
Jonga ileyibhile yomhla wokwenziwa kunye nokusetyenziswa.
Emva kokuvulwa kokuqala, i-reagent inokugcinwa kwi -20 ± 5 ° C ngaphandle kwenyanga enye okanye kude kube sekupheleni kwexesha le-reagent, nokuba nguwuphi na umhla oza kuqala, ukunqanda imijikelo ephindaphindiweyo yokuqhaqha, kunye nenani lokukhenkceza kwe-reagent. Imijikelo yokunyibilika akufuneki idlule amaxesha ama-6.
【Applicable instrument】ABI 7500, SLAN-96P, Roche-LightCycler-480.
【Sample requirements 】
1.Uhlobo lwesampulu esebenzayo: Isisombululo se-nucleic acid ekhutshweyo.
2.Ukugcinwa kwesampuli kunye nokuthutha: Gcina kwi-20 ± 5 ℃ kwiinyanga ezi-6. Khulisa kwaye unyibilikise iisampuli ngaphandle kwamaxesha angama-6.
【TEsting metihod】
1.Nucleic acid extraction
Khetha ikhithi efanelekileyo yokukhupha i-nucleic acid ukukhupha i-viral nucleic acid, kwaye ulandele imiyalelo yekhithi ehambelanayo. Kunconywa ukusebenzisa i-Nucleic acid extraction kunye nekiti yokucoca eveliswe yi-Yixin Bio-Tech (Guangzhou) Co., Ltd. okanye ikiti yokucoca i-nucleic acid efanayo.
2. Reaction reagent preparation
2.1 For BST-SARS-25:
( 1) Susa umxube we-SARS-Cov-2 kunye noMxube we-Enzyme, unyibilike ngokupheleleyo kumxube wobushushu begumbi ngokucokisekileyo ngesixhobo seVortex kwaye emva koko ube yi-centrifuge ngokufutshane.
(2) 16.5uL Umxube we-Enzyme wongezwa kwi-358.5uL SARS-Cov-2 Mix kwaye yaxutywa ngokucokisekileyo ukufumana isisombululo sokusabela okuxubileyo.
(3) Lungiselela ityhubhu ye-octal ye-0.2 mL ye-PCR kwaye uyiphawule nge-15uL yesisombululo sempendulo exutywe ngasentla kwiqula ngalinye.
(4) Yongeza i-15 μL yesisombululo se-nucleic acid ecocekileyo, ulawulo oluhle kunye nolawulo olubi, kwaye ugubungele ngononophelo i-octal tube cap.
(5) Xuba kakuhle ngokujikela phantsi, kwaye ngokukhawuleza i-centrifuge ugxininise ulwelo ezantsi kombhobho.
1
2.2 For BST-I-SARS-DR-25:
( 1) Yongeza i-375ul 2x Buffer ku-SARS-Cov-2 Mix((Lyophilised) ukulungisa umxube wokusabela. Xuba ngocoselelo ngombhobho kwaye emva koko ube yi-centrifuge ngokufutshane.. （Xa umxube wokusabela ulungisiwe, kuyacetyiswa ukuba ugcinwe ku -20℃ ukugcinwa kwexesha elide.)
(2) Lungisa ityhubhu ye-octal ye-0.2 mL ye-PCR kwaye uyiphawule nge-15μL yomxube wokuphendula kakuhle.
(3) Yongeza i-15μL yesisombululo se-nucleic acid ecocekileyo, ulawulo oluhle kunye nolawulo olubi, kwaye ugubungele ngononophelo i-octal tube cap.
(4) Xuba kakuhle ngokujikela phantsi, kwaye ngokukhawuleza i-centrifuge ukugxininisa ulwelo ezantsi kombhobho.
3. I-PCR ampubomication (Nceda ujonge kwisixhobo sokwenza iisetingi zokusebenza.）
3. I-1 Beka i-PCR 8-tube kwigumbi lesampula lesixhobo se-PCR se-fluorescent, kwaye usethe isampuli ukuba ihlolwe, ulawulo oluhle kunye nolawulo olubi ngokomyalelo wokulayisha.
3.2 Ijelo lokubona iFluorescence:
( 1) I-ORF1ab i-gene ikhetha i-channel yokufumanisa i-FAM (Ingxelo: FAM, Quencher: Akukho).
(2) I-N gene ikhetha i-channel yokufumanisa i-VIC (Ingxelo: VIC, Quencher: Akukho).
(3) I-gene yomgangatho wangaphakathi ikhetha umjelo wokufumanisa we-CY5 (Ingxelo: CY5, Quencher: Akukho).
(4) I-Passive Reference isetelwe ku-ROX.
3.3 PCR useto lweparamitha yeprogram:
| Inyathelo | Iqondo lobushushu(℃) | Ixesha | Inani lemijikelo | |
| 1 | Reverse transcription reaction | 50 | 15 imiz | 1 |
| 2 | Ukuqaliswa kwe-enzyme ye-Taq | 95 | 2.5 imiz | 1 |
| 3 | Ukuqaliswa kwe-enzyme ye-Taq | 93 | 10 s | 43 |
| Ukwandiswa kwe-Anealing kunye nokufunyanwa kwe-fluorescence | 55 | 30 s | ||
Emva kokuseta, gcina ifayile kwaye uqhube inkqubo yokusabela.
4.Results analysis
Emva kokuba inkqubo iphelile, iziphumo zigcinwa ngokuzenzekelayo, kwaye i-curve yokukhulisa ihlalutye. Igophe lokukhulisa isetyenziselwe kwisixhobo esingagqibekanga somda.
【Explanation of test resuiilt 】
1. Qinisekisa ukufaneleka kovavanyo: Ulawulo olulungileyo lwe-FAM, i-VIC ishaneli kufuneka ibe ne-curve ye-amplification eqhelekileyo, kwaye ixabiso le-Ct ngokuqhelekileyo lingaphantsi kwe-34, kodwa lingaguquguquka ngenxa yemimiselo eyahlukeneyo yezixhobo ezahlukeneyo. Ulawulo olubi lwe-FAM, i-VIC channel kufuneka ibe yi-Ct enganyuswanga. Kuvunyelwene ukuba ezi mfuno zingentla kufuneka zifezekiswe ngexesha elinye, ngaphandle koko olu vavanyo aluvumelekanga.
2. Iziphumo zesigwebo
| FAM/VIC channel | Isiphumo somgwebo |
| Ct＜37 | Uvavanyo lwesampulu lunayo |
| 37≤Ct＜40 | Igophe lokukhulisa limile okwe-S, kwaye iisampulu ezikrokrisayo kufuneka ziphinde zihlolwe; ukuba iziphumo zokuphononongwa kwakhona ziyahambelana, zigwetywa njengezilungileyo, ngaphandle koko zibi |
| Ct≥40 Okanye Akukho Ukukhulisa | Uvavanyo lwesampulu alulunganga (okanye ngaphantsi komda osezantsi wokubhaqwa kwekhithi) |
Qaphela: ( 1) Ukuba omabini itshaneli ye-FAM kunye nejelo le-VIC zinethemba ngaxeshanye, i-SARS-Cov-2 izimisele ukuba ibe ntle.
(2) Ukuba itshaneli ye-FAM okanye i-VIC ishaneli ilungile kwaye elinye ijelo libi, uvavanyo kufuneka luphindwe. Ukuba une-positive kwangaxeshanye, iya kugwetywa njenge-SARS-Cov-2 positive, kungenjalo iya kugwetywa njenge-SARS-Cov-2 negative.
Iimveliso ezinxulumeneyo
-
Ikhithi yokuFumana i-COVID-19 ye-IgM/IgG ye-Antibody
I-COVID-19 IgM/IgG yeKhithi yokuFumana i-Antibody ye-COVID-19 (iNdlela ye-Colloidal yeGolide ye-Immunochromatography) Incwadi yeMveliso yeMveliso 【Igama leMveliso】 COVID-19 IgM/IgG Ikhithi yokuFumana i-Antibody ye-Colloidal yeGolide ye-Immunochromatography) 【UVAVANYO UVAVANYO/I-PACKAGING EC EC,I-PACKAGING 1. 【 ISISHWANKATHELO】 Inoveli coronaviruses zezohlobo lwe-β. I-COVID-19 sisifo esosulelayo esinamandla sokuphefumla. Ngokuqhelekileyo abantu basengozini. Okwangoku, abaguli bosulelwe yinoveli coronavirus ngowona mthombo uphambili ...
-
I-Nucleic Acid Extraction okanye iKhiti yokuCoca
I-Nucleic Acid Extraction Okanye I-Purification Kit okanye igcinwe ku -20℃. Isampulu kufuneka ithuthwe kusetyenziswa i-0℃ curling. Intshayelelo I-Nucleic Acid Extraction Okanye IKit yokuCoca (iNdlela yeMagnetic Beads) yenzelwe ukucocwa ngokuzenzekelayo kwe-RNA kunye ne-DNA kulwelo lomzimba (olufana ne-swabs, i-plasma, i-serum) kusetyenziswa izixhobo zokutsalwa kwe-nucleic acid ezizenzekelayo. Itekhnoloji ye-Magnetic-particle ibonelela nge-DNA/RNA ekumgangatho ophezulu elungele ...
-
SARS-CoV-2 Antigen Assay Kit (Immunochromatogr...
SARS-CoV-2 Antigen Assay Kit (Immunochromatography Method) Incwadi yeMveliso 【IGAMA LEMVELISO】SARS-CoV-2 Antigen Assay Kit （Immunochromatography Method) INGCACISO YOKUPAKISHA】 1 Uvavanyo/iKit noveli ye-ABS ye-coronavirus I-COVID-19 sisifo esosulelayo esinamandla sokuphefumla. Ngokuqhelekileyo abantu basengozini. Okwangoku, abaguli bosulelwe yinoveli coronavirus ngabona baphambili ...
-
SARS-CoV-2 Antigen Assay Kit
SARS-CoV-2 Antigen Assay Kit (Immunochromatography Method) Incwadi yeMveliso 【IGAMA LEMVELISO】SARS-CoV-2 Antigen Assay Kit （Immunochromatography Indlela) 【IINGCACISO YOKUFAKA】 1 Uvavanyo/IKit 1 ABT/Kit 1 ABT/Kit 1 ABET 】 I ii-coronavirus ezintsha zezodidi lwe-β. I-COVID-19 sisifo esosulelayo esinamandla sokuphefumla. Ngokuqhelekileyo abantu basengozini. Okwangoku, izigulana ezosulelwe yinoveli coronavirus ngoyena mthombo wosulelo; abantu abasulelekileyo abangenazimpawu...