Product Detail
World Health Organisation (WHO) inotungamira kuti antigen-inoona nekukurumidza diagnostic bvunzo (Ag-RDRs) inogona kupa inokurumidza uye isingadhure nzira yekuongorora inoshanda SARS-CoV-2 utachiona pane nucleic acid amplification bvunzo (NAATs), uye WHO inokurudzirawo. kuti maAg-RDT anosangana nedikidiki mashandiro ekuita anogona kushandiswa kutarisisa nyaya dzekutanga, kutsvaga kuonana, panguva yekuferefeta kwekubuda uye kuongorora mafambiro echirwere munharaunda.
Features
● Zvakakwana:Matatu ejini anoona mune imwe bvunzo
● inowirirana:Adaptive kune zvakajairika zvishandiso neCY5, FAM, VIC/HEX chiteshi.
● Kuita kwakanakisa:Kunzwa kwakanyanya uye kujeka, LOD = 200 makopi / ml.
Technical Parameter
| Packing Specification | 50 bvunzo/kiti, zana bvunzo/kiti |
| Target Region | ORF1ab, N, E |
| Inoshanda Sample | Sputum, Oropharyngeal Swab |
| Muganhu Wekuonekwa | 200 makopi / ml |
| Total coincidence Rate | 99.55% |
| Ct kukosha (CV,%) | ≤5.0% |
| Positive Coincidence Rate | 99.12% |
| Negative Coincidence Rate | 100% |
| Mamiriro Ekuchengetedza uye Zuva rekupera | Yakachengetwa pa -20±5 ℃, uye inoshanda kwenguva pfupi kwemwedzi gumi nemiviri. |
| Kudzora Kwemukati | Ehe |
| Catalog Number | A7793YF-50T, A7793YF-100T |
| Certification | CE |
| Sayenzi | Nasopharyngeal swab, Oropharyngeal swab, Alveolar lavage fluid, Mate uye sputum |
| Applicable Instrument | ABI 7500, Roche Light Cycler 480Ⅱ , Roche Cobas z 480, SLAN-96P Chaiyo-Nguva PCR System |
Test Procedure
1. Nucleic Acid Extraction
Kushanda kunofanirwa kuitwa maererano nebhuku rekutora kit.
2. Kugadzirira System:
1) Bvisa reagent uye nyungudutsa reagent zvachose.Pindura musanganiswa uye centrifuge ipapo.N bvunzo maitiro (N = nhamba yemasamples anoedzwa + positive control + negative control + 1) akagadzirirwa maitiro masisitimu, zvichiteerana, sezvinotevera.
| Vomponentss | Vhoriyamu ye1 reaction system | Vhoriyamu yeN reaction system |
| Nucleic acid amplification reaction solution Mix (A7793YF) | 18µL | 18µL *N |
| Enzyme musanganiswa | 2 µL | 2µL *N |
| Vhoriyamu yese | 20 μL | 20µL *N |
2) Rection distribution: Mhinduro yemhinduro yakasanganiswa uye centrifuged, uye imwe neimwe chubhu yakagoverwa muhuwandu hwe20μL muPCR tube yakakodzera fluorescence PCR apparatus.
3. Loading
5μL yeyakabudiswa sampuli nucleic acid, positive control nucleic acid uye negative control nucleic acid inowedzerwa kune maitiro ekugadzirisa, uye huwandu hwekuita vhoriyamu i25μL.Sunga chivharo chechubhu uye uiendese kunzvimbo yekuyedza yeamplification mushure memasekondi mashoma ecentrifugation.
4. PCR Amplification Assay
1) Isa iyo PCR reaction chubhu mune fluorescent PCR amplification chiridzwa chekusimudza kuona.
2) Cycle parameter setting:
| Chirongwa | Nhamba yezvikamu | Tembiricha | Reaction nguva | |
| 1 | 1 | 50 ℃ | 10 min | |
| 2 | 1 | 95℃ | 30 sec | |
| 3 | 45 | 95℃ | 5 sec | |
| 60 ℃ | 30 sec | Fluorescence Collection | ||
3) Zvirongwa zvekuonekwa:
Nzira dzekuona dzakaiswa kuFAM, VIC, ROX uye CY5, inoenderana neORF1ab, N gene, uye E Gene, RNase P yemukati kudzora, zvichiteerana."Quencher Dye" uye "Passive Reference" yakaiswa ku "Hapana" yeABI 7500 chiridzwa.Isa iyo Positive Control, Negative Control, uye Sample (Isingazivikanwe) kuitira kuti masampuli anoenderana, uye isa zita remuenzaniso mu "Sample Name" column.
Kune X-POCH16, kushanda uye chirongwa ndeichi:
1) Mushure mekuzviongorora kwapera, vhura chivharo uye isa PCR reaction machubhu munzvimbo dzakatarwa muchiridzwa.
2) Tanga nekusarudza iyo "Exper."sarudzo.Sarudza iyo "Yese" sarudzo kana nemaoko sarudza nzvimbo yekuita kuruboshwe rwechidzitiro.
3) Sarudza "LOAD" sarudzo;sarudza chirongwa chekuedza;tinya "KUITA" uye "RUN".Iyo purogiramu inotora 30min42s kuti ipedze.
Nzira dzekuona dzeDefault program dzakaiswa kuFAM, VIC, ROX uye CY5, inoenderana neORF1ab, N gene, uye E Gene, RNase P yemukati kudzora, zvichiteerana.
Iyo yekutenderera parameter yeDefault chirongwa ndeiyi inotevera:
| Chirongwa | Nhamba ye | Tembiricha | Reaction nguva |
| 1 | 1 | 50 ℃ | 2min |
| 2 | 1 | 95℃ | 30sec |
| 3 | 41 | 95℃ | 2sec |
| 60 ℃ | 13sec | Fluorescence |
5. Threshold Setting
Zvinoenderana nemufananidzo wakaongororwa, gadzirisa kukosha kweKutanga, Kupera kukosha kweBaseline uye Threshold kukosha (Kutanga kukosha uye kukosha kwekupedzisira kunokurudzirwa kuiswa kuve 3 uye 15 zvakateerana, uye iyo amplification curve yeakaipa control inogadziriswa kuti ive flat kana yakaderera pane chikumbaridzo Mutsetse), tinya Ongororo kuti uwane otomatiki ongororo iyo yemuenzaniso Ct kukosha.Wona zvawanikwa muReport interface.
6. Quality Control Standard
Kudzora kwega kwega kwekiti kunofanirwa kuzadzisa zvinodiwa zvinotevera ne 'S' curve, zvikasadaro kuedza hakushande.
| Kuona nzira | Negative control | Positive control |
| FAM(ORF1ab) | Kwete Ct | Ct≤38 |
| VIC(N) | Kwete Ct | Ct≤38 |
| ROX(E) | Kwete Ct | Ct≤38 |
| CY5(RP) | Kwete Ct | Ct≤38 |
【Kuchekwa Kukosha】
Zvinoenderana nemhedzisiro ye100 oropharyngeal swab samples uye zana sputum samples, uye neROC curve nzira, iyo Cut-off kukosha kweOFR1ab, N gene E Gene yekiti iyi ndeye Ct = 38.
FAQ
Mune kit iyi, maprimers uye maprobes eiyo chaiyo-nguva fluorescent PCR tekinoroji akagadzirirwa akachengetwa uye chaiwo matunhu eORF1ab, N uye E gene ye2019-nCoV zvakateerana.Munguva yePCR kukwidziridzwa, probe inosungirira kune template, uye iyo 5'-yekupedzisira mutori wenhau boka reiyo probe yakatsemurwa neiyo Taq enzyme (5'→ 3' exonuclease chiitiko), nekudaro ichibva kure neboka rekudzima kuti igadzire chiratidzo chefluorescent. .Iyo chaiyo-nguva amplification curve inorongwa otomatiki zvichibva pane yakaonekwa fluorescence chiratidzo, uye sampuli Ct kukosha inoverengerwa.FAM, VIC uye ROX fluorophores zvakanyorwa kuORF1ab gene,N gene uye E gene probes.Nekushandisa bvunzo imwe chete, kuongororwa kwemhando yemajini matatu ari pamusoro e2019-nCoV anogona kuitwa panguva imwe chete.
Iyo kit inopihwa yemukati yekudzora yakanangana neRNase P gene yekutarisa kliniki samples kuunganidzwa, kubata, kudhirowa uye RT-PCR maitiro kudzivirira manyepo-asina kunaka mhedzisiro.Iyo yemukati kutonga yakanyorwa neCY5 fluorescent boka.
1. Ongorora uye kududzira bvunzo mhinduro kana chiridzwa chakajairika, uye kutonga kwakanaka, kutonga kwakashata uye mukati mekutonga bvunzo mhinduro zvinosangana nemhando yekudzora mwero.
2. Iyo amplification curve yekutonga kwemukati (CY5) inoratidza chimiro cheS curve uye Ct ≤ 38, kududzira mhedzisiro yemajini anotangwa inoenderana nemamiriro anotevera.
| Kuona nzira | Kuturikira mhedzisiro yemajini anonangwa | ||
| FMA | VIC (N gene) | ROX (E gene) | |
| Ct≤38 | Ct≤38 | Ct≤38 | Iine yakajairwa S amplification curve, iyo Ct kukosha ndeye≤38, inoenderana chinangwa geni yakanaka. |
| 38 ＜ Ct ＜ 40 | 38 ＜ Ct ＜ 40 | 38 ＜ Ct ＜ 40 | Iine yakajairwa S amplification curve, edzazve inoenderana chinangwa gene remuenzaniso zvakare. Kana iyo Ct yakakosha ＜ 40 ine yakajairwa S amplification curve, inoenderana inotangwa gene yakanaka;kana iyo Ct yakakosha≥40, iyo inoenderana neiyo gene gene haina kunaka |
| Ct≥40 | Ct≥40 | Ct≥40 | Iyo inoenderana nejini yechinangwa haina kunaka |
Dudziro yemhedzisiro ye2019-nCoV:
Zvinoenderana nemhedzisiro yeORF1ab, N gene uye E gene, dudziro sezvinotevera:
1) Kana maviri kana MATATU majini emajini akaonekwa ari akanaka, iyo 2019-nCoV yakanaka;
2) Kana IMWE CHETE kana KUSINA yemajini akaonekwa ari akanaka, iyo 2019-nCoV haina kunaka.
Ongorora: Iyo yekukwiridzira curve yeyakanaka sampuli inofanirwa kunge iine yakajairwa S curve.Nekudaro, kana chinangwa chekutarisa chakanyanya, iyo yemukati mwero kutonga inogona kusakwidziridzwa uye sampuli inogona kutongerwa zvakananga seyakanaka.Kana chero maviri emajini ekunangwa akawana Ct≤38, iyo 2019-nCoV yakanaka.Kana chero maviri emajini ekunangwa akawana Ct≥40, iyo 2019-nCoV haina kunaka.Kana Ct≥40, kana kuratidza kusakosha, kududzira mhedzisiro yejini yechinangwa haina kunaka.
3. Kana ese maCt values eFAM, VIC, ROX neCy5 machanera anopfuura makumi matatu nemasere kana pasina ari pachena akajairika S amplification curve:
1) Pane / pane zvinhu (s) mumuenzaniso izvo zvinotadzisa PCR kuita.Inokurudzirwa kudzikisa sampuli kuti iongororwe zvakare.
2) Iyo nzira yekubvisa nucleic acid haina kujairika, saka inokurudzirwa kuburitsa zvakare.
nucleic acid yekuongororazve.
3) Sample iyi yanga isiri sampuli yakakodzera panguva yesampling, kana kuderedzwa
panguva yekufambisa uye kuchengetedza.
Ndidzo nzira mbiri dzatinogona kuona mamiriro aya: NAAT neAntigen.
(Inobva kuCDC yeLos Angeles)
