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
【Producet name 】New Coronavirus(SARS-Cov-2) Nucleic Acid Detection Kit(Fluorescent RT-PCR Probe Method)
【Packaging specifications 】25 Miedzo/Kit
【Intended usage】
Kiti iyi inoshandiswa kucherechedzwa kwemhando ye nucleic acid kubva kune nyowani coronavirus mu nasopharyngeal swabs, oropharyngeal (huro) swabs, anterior nasal swabs, yepakati-turbinate swabs, mhino dzekugezesa uye mhino kushuvira kubva kune vanhu vanofungidzirwa kuti vane COVID19 nemupi wehutano. Kuonekwa kweORF1ab neN majini eiyo coronavirus nyowani inogona kushandiswa kubatsiridza kuongororwa uye epidemiological yekutarisa hutachiona hutsva hwekoronavirus.
【Principles of the procedure 】
Iyi kit yakagadzirirwa chaiyo TaqMan probes uye chaiwo maprimers akagadzirirwa iyo novel coronavirus (SARS-Cov-2) ORF1ab uye N gene sequences. Iyo PCR reaction solution ine 3 seti yeakananga maprimers uye fluorescent probes yekunyatso tariswa kwezvinangwa, uye imwe seti yemamwe maprimers uye fluorescent probes inoshandiswa seyemukati mwero kutonga kwekiti yekuona endogenous ekuchengetedza imba.
Nheyo yekuyedza ndeyekuti iyo chaiyo fluorescent probe inogayiwa uye yakaderedzwa neiyo exonuclease chiitiko cheTaq enzyme muPCR reaction, kuitira kuti mutori wenhau fluorescent boka uye rakadzimwa fluorescent boka riparadzaniswe, kuitira kuti fluorescence monitoring system igamuchire fluorescent. chiratidzo, uyezve Kuburikidza nekupfumisa mhedzisiro yePCR kukwidziridzwa, iyo fluorescence chiratidzo cheiyo probe inosvika payakaiswa chikumbaridzo kukosha-Ct kukosha (Cycle threshold). Munyaya yekusina chinangwa amplicon, boka revatori venhau reprobe riri padyo neboka rekudzima. Panguva ino, iyo fluorescence resonance energy transfer inoitika, uye fluorescence yeboka remutori wenhau inodzimwa neboka rekudzima, kuitira kuti chiratidzo chefluorescent chirege kuonekwa nechiridzwa chePCR chefluorescent.
Kuti utarise kushandiswa kwemareagents panguva yekuyedzwa, kit inoshongedzerwa neakanaka uye asina kunaka kudzora: iyo yakanaka kutonga ine inotangwa saiti recombinant plasmid, uye iyo yakaipa kutonga iDistilled mvura, iyo inoshandiswa kutarisa kusvibiswa kwezvakatipoteredza. Inokurudzirwa kuisa hutungamiri hwakanaka uye hutongi husina kunaka panguva imwe chete pakuedza.
【Main components 】
| Cat. No. | BST-SARS-25 | BST-SARS-DR-25 | Components | |
| Name | Specification | Quantity | Quantity | |
| Positive control | 180 μL / bhodhoro | 1 | 1 | Yakagadzirwa plasmids, Distilled mvura |
| Negative control | 180 μL / bhodhoro | 1 | 1 | Distilled mvura |
| SARS-Cov-2 Mix | 358.5 μL/mudziyo | 1 | / | Yakananga maprimer pairs, chaiyo yekuona fluorescent probes, dNTPs, , MgCl2, KCl, Tris-Hcl, Distilled mvura, nezvimwe. |
| Enzyme Mix | 16.5 μL / vial | 1 | / | Taq enzymes, reverse transcriptase,UNG enzymes, nezvimwe. |
| SARS-Cov-2 Mix(Lyophilised) | 25 miedzo / vial | / | 1 | Yakananga primer pairs, chaiyo yekuona fluorescent probes, dNTPs, Taq enzymes, reverse transcriptase, Distilled mvura, nezvimwe. |
| 2x Buffer | 375 μL/mudziyo | / | 1 | MgCl2, KCl, Tris-Hcl, Distilled mvura, nezvimwewo. |
Cherechedza:(1) Izvo zvikamu mumaketsi akasiyana-siyana haagone kusanganiswa kana kuchinjana.
(2) Gadzirira yako reagent: Nucleic acid yekubvisa kit.
【Storage conditions uye expiration date 】
For BST-SARS-25:Kutakura uye chitoro pa -20±5 ℃ kwenguva yakareba.
For BST-SARS-DR-25:Kutakura pakamuri tembiricha. Chengetedza pa -20±5 ℃ kwenguva yakareba.
Dzivisa kudzokororwa kwechando-kunyungudika kutenderera. Nguva yechokwadi inotarirwa 12months.
Ona chinyorwa chezuva rekugadzirwa uye kushandiswa.
Mushure mekuvhurwa kwekutanga, reagent inogona kuchengetwa pa -20 ± 5 ° C kweisingapfuure 1 mwedzi kana kusvika pakupera kweiyo reagent nguva, chero zuva rinouya pekutanga, kudzivirira kudzokororwa kwechando-kunyorova kutenderera, uye huwandu hwekuomesa chando. -thaw cycles haifaniri kudarika 6 nguva.
【Applicable instrument】ABI 7500, SLAN-96P, Roche-LightCycler-480.
【Sample requirements 】
1.Inoshanda muenzaniso wemhando: Yakabviswa nucleic acid solution.
2.Sample kuchengetedza uye kutakura: Chengetedza pa-20 ± 5 ℃ kwemwedzi 6. Isai uye nyura masampuli kwete kupfuura 6 nguva.
【Testing method】
1.Nucleic acid extraction
Sarudza yakakodzera nucleic acid yekubvisa kit kuti ubvise viral nucleic acid, uye tevera inoenderana kit mirairo. Zvinokurudzirwa kushandisa Nucleic acid yekubvisa uye kit yekuchenesa inogadzirwa neYixin Bio-Tech (Guangzhou) Co., Ltd. kana yakaenzana nucleic acid yekuchenesa kit.
2. Reaction reagent preparation
2.1 For BST-SARS-25:
( 1) Bvisa iyo SARS-Cov-2 Musanganiswa uye Enzyme Musanganiswa, nyungudika zvachose pakamuri tembiricha musanganise zvakakwana neVortex mudziyo uyezve centrifuge muchidimbu.
(2) 16.5uL Enzyme Mix yakawedzerwa ku358.5uL SARS-Cov-2 Mix uye yakasanganiswa zvakakwana kuti iwane mhinduro yakasanganiswa.
(3) Gadzirira yakachena 0.2 mL PCR octal chubhu woimaka ne 15uL yemusanganiswa wekuita mhinduro patsime.
(4) Wedzera 15 μL yeakacheneswa nucleic acid mhinduro, iyo yakanaka kutonga uye iyo yakaipa kutonga, uye nyatso vhara octal chubhu chivharo.
(5) Sanganisa zvakanaka nekupidigura wakatarisa pasi, uye nekukasira centrifuge kuisa mvura pazasi pechubhu.
1
2.2 For BST-SARS-DR-25:
( 1) Wedzera 375ul 2x Buffer kuSARS-Cov-2 Mix((Lyophilised) kugadzirira musanganiswa wekuita. Sanganisa zvakanyanyisa nepipetting uye ipapo centrifuge zvishoma. kuchengetedza kwenguva refu.)
(2) Gadzirira yakachena 0.2 mL PCR octal chubhu woimaka ne 15μL ye reaction mix patsime.
(3) Wedzera 15μL yeakacheneswa nucleic acid mhinduro, iyo yakanaka kutonga uye iyo yakaipa kutonga, uye nyatso vhara octal chubhu chivharo.
(4) Sanganisa zvakanaka nekupindurudza wakatarisa pasi, uye nekukurumidza centrifuge kuisa pfungwa pamvura pazasi pechubhu.
3. PCR amplification (Ndokumbira utarise kune bhuku rekushandisa rekushandisa marongero.）
3. 1 Isai PCR 8-tube mukamuri yemuenzaniso yefluorescent PCR chiridzwa, uye isa muenzaniso kuti uongororwe, kutonga kwakanaka uye kutonga kusina kunaka maererano nekurongeka kwekutakura.
3.2 Fluorescence yekuona chiteshi:
(1) ORF1ab gene inosarudza nzira yekuona yeFAM (Mutori wenhau: FAM, Quencher: Hapana).
(2) N gene inosarudza nzira yekuona yeVIC (Mutori wenhau: VIC, Quencher: Hapana).
(3) Internal standard gene inosarudza nzira yekuona yeCY5 (Mutori wenhau: CY5, Quencher: Hapana).
(4) Iyo Passive Reference yakaiswa kuROX.
3.3 PCR chirongwa parameter kumisikidza:
| Danho | Tembiricha(℃) | Nguva | Nhamba yezvikamu | |
| 1 | Reverse transcription reaction | 50 | 15 min | 1 |
| 2 | Taq enzyme activation | 95 | 2.5 min | 1 |
| 3 | Taq enzyme activation | 93 | 10 p | 43 |
| Anealing yekuwedzera uye fluorescence kuwana | 55 | 30 p | ||
Mushure mekugadzirisa, chengetedza faira uye shandisa purogiramu yekuita.
4.Results analysis
Mushure mekunge chirongwa chapera, mibairo inochengetwa otomatiki, uye iyo amplification curve inoongororwa. Iyo amplification curve yakaiswa kune chiridzwa default chikumbaridzo.
【Explanation of test results 】
1. Sarudza chokwadi chekuedza: The positive control FAM, VIC channel inofanira kuva neakajairika amplification curve, uye kukosha kweCt kunowanzova pasi pe34, asi inogona kuchinja nekuda kwezvisungo zvakasiyana-siyana zvezviridzwa zvakasiyana. Iyo isina kunaka inodzora FAM, VIC chiteshi haifanirwe kunge isina-amplified Ct. Zvinotenderwa kuti zvinodiwa pamusoro apa zvinofanirwa kusangana panguva imwe chete, zvikasadaro bvunzo iyi haina basa.
2. Mutongo wekutonga
| FAM/VIC chiteshi | Mugumisiro wekutonga |
| Ct＜37 | Sample test is positive |
| 37≤Ct＜40 | Iyo amplification curve ndeye S-yakaita, uye masampuli anofungidzira anoda kuongororwazve; kana mhinduro dzekuongorora zvakare dzichienderana, inotongwa seyakanaka, kana zvisina kudaro haina kunaka |
| Ct≥40 Kana Hapana Amplification | Sample test haina kunaka (kana pazasi peiyo yakaderera muganho wekuona kit) |
Ongorora: (1) Kana ese ari maviri chiteshi cheFAM uye VIC chiteshi zvakanaka panguva imwe chete, iyo SARS-Cov-2 yakatsunga kuve yakanaka.
(2) Kana iyo FAM chiteshi kana VIC chiteshi chakanaka uye imwe chiteshi haina, bvunzo inofanira kudzokororwa. Kana iri positive panguva imwe chete, inozotongwa seSARS-Cov-2 yakanaka, zvikasadaro ichatongwa seSARS-Cov-2 isina kunaka.
Zvigadzirwa zvinoenderana
-
COVID-19 IgM/IgG Antibody Detection Kit
COVID-19 IgM/IgG Antibody Detection Kit (Colloidal Gold Immunochromatography Method) Chigadzirwa Chinyorwa 【PRODUCT ZITA】 COVID-19 IgM/IgG Antibody Detection Kit （Colloidal Gold Immunochromatography Method) 【TsKiidzo Muedzo/Kuongorora/Kuongorora 1. 【 ABSTRACT】 Iyo novel coronaviruses ndeye β genus. COVID-19 chirwere chekufema chinotapukira. Vanhu vanowanzo batwa nedambudziko iri. Parizvino, varwere vakatapukirwa neiyo novel coronavirus ndiyo sosi huru ...
-
Nucleic Acid Kubvisa Kana Kuchenesa Kit
Nucleic Acid Kubvisa Kana Kuchenesa Kit kana kuchengetwa pa -20 ℃. Muenzaniso unofanirwa kutakurwa uchishandisa 0 ℃ curling. Nhanganyaya Iyo Nucleic Acid Kubvisa Kana Kuchenesa Kit (Magnetic Beads Method) yakagadzirirwa kucheneswa otomatiki kweRNA neDNA kubva kumvura dzemuviri (senge swabs, plasma, serum) uchishandisa otomatiki nucleic acid yekubvisa zviridzwa. Magnetic-particle tekinoroji inopa yemhando yepamusoro DNA/RNA inokodzera ...
-
SARS-CoV-2 Antigen Assay Kit (Immunochromatogr...
SARS-CoV-2 Antigen Assay Kit (Immunochromatography Method) Chigadzirwa Chinyorwa 【PRODUCT ZITA】SARS-CoV-2 Antigen Assay Kit (Immunochromatography Method) 【KUPAKA ZVINOSUNGIRWA】 1 Bvunzo/Kit norubheti ndeyeiyo β ACT genus】 COVID-19 chirwere chekufema chinotapukira. Vanhu vanowanzo batwa nedambudziko iri. Parizvino, varwere vakatapukirwa neiyo novel coronavirus ndivo vakuru ...
-
SARS-CoV-2 Antigen Assay Kit
SARS-CoV-2 Antigen Assay Kit (Immunochromatography Method) Chigadzirwa Chinyorwa 【PRODUCT ZITA】SARS-CoV-2 Antigen Assay Kit (Immunochromatography Method) 【KUPAKA ZVINOTAURWA】 1 Muedzo/Ts 10Kit 1 ABT/Kit 1 ABT ABT/2Kit 1 ABT ABT/2Kit 】 The novel coronaviruses ndeye β genus. COVID-19 chirwere chekufema chinotapukira. Vanhu vanowanzo batwa nedambudziko iri. Parizvino, varwere vakatapukirwa neinoveli coronavirus ndiyo honzeri yehutachiona; vanhu vane asymptomatic vane hutachiona ...