Cikakken Bayani
Hukumar Lafiya ta Duniya (WHO) ta ba da umarnin cewa gwaje-gwajen saurin gano maganin antigen (Ag-RDRs) na iya ba da hanya mai sauri da ƙarancin tsada don tantance kamuwa da cutar SARS-CoV-2 mai aiki fiye da gwajin haɓaka haɓakar acid nucleic (NAATs), kuma WHO ta kuma ba da shawarar. za a iya amfani da Ag-RDTs da suka cika mafi ƙarancin buƙatun aiki don gano shari'ar farko, gano tuntuɓar juna, yayin binciken fashewa da kuma sa ido kan abubuwan da ke faruwa na kamuwa da cuta a cikin al'ummomi.
Siffofin
● M:Kwayoyin da aka yi niyya guda uku sun gano a cikin gwaji ɗaya
● masu jituwa:Mai dacewa da kayan aiki gama gari tare da tashoshin CY5, FAM, VIC/HEX.
● Kyakkyawan aiki:Babban hankali da ƙayyadaddun bayanai, LOD = 200 kwafi / ml.
Sigar Fasaha
| Ƙimar tattarawa | Gwaje-gwaje 50/kit, gwaji 100/kit |
| Yankin Target | ORF1ab, N, E |
| Samfurin da ya dace | Sputum, Oropharyngeal Swab |
| Iyakar Ganewa | 200 kwafi/ml |
| Jimlar Ƙimar Daidaituwa | 99.55% |
| Ƙimar Ct (CV,%) | ≤5.0% |
| Matsakaicin Daidaito Mai Kyau | 99.12% |
| Matsakaicin Rashin daidaituwa | 100% |
| Yanayin Ajiya da Ranar Karewa | An adana shi a -20± 5℃, kuma yana aiki na ɗan lokaci na watanni 12. |
| Ikon Cikin Gida | Ee |
| Lambar Catalog | Saukewa: A7793YF-50T |
| Takaddun shaida | CE |
| Samfura | Nasopharyngeal swab, Oropharyngeal swab, Alveolar lavage ruwa ruwa, Saliva da sputum |
| Abubuwan da ake Aiwatar da su | ABI 7500, Roche Light Cycler 480Ⅱ , Roche Cobas z 480, SLAN-96P Tsarin PCR na Gaskiya |
Tsarin Gwaji
1. Ciwon Acid Nucleic
Ya kamata a gudanar da aiki bisa ga littafin kayan aikin hakar.
2. Tsari Tsari:
1) Fitar da reagent kuma a narke reagent gaba daya.Juya cakuda da centrifuge nan da nan.N gwajin halayen (N = adadin samfuran da za a gwada + kulawa mai kyau + iko mara kyau + 1) an shirya don tsarin amsawa, bi da bi, kamar haka.
| Voponents | Ƙarar don tsarin amsawa 1 | Ƙarar don tsarin amsawa N |
| Nucleic acid amplification reaction Mix (A7793YF) | 18 µl | 18µL* N |
| Enzyme cakuda | 2 µl | 2 µL * N |
| Jimlar girma | 20 µl | 20µL* N |
2) Rarraba ra'ayi: Maganin amsawa ya gauraye kuma an ba da shi, kuma an ba da kowane bututu a cikin adadin 20μL a cikin bututun PCR wanda ya dace da na'urar PCR mai haske.
3. Loading
5μL na samfurin nucleic acid da aka fitar, ingantaccen iko nucleic acid da kuma mummunan iko nucleic acid ana kara su zuwa tsarin amsawa, kuma jimlar amsawar shine 25μL.A ɗaure murfin bututu kuma matsar da shi zuwa wurin gwajin haɓakawa bayan ƴan daƙiƙa na ɗaki.
4. PCR Amplification Assay
1) Sanya bututun amsawar PCR a cikin kayan haɓakawa na PCR mai kyalli don gano haɓakawa.
2) Saitin siginar zagayowar:
| Shirin | Yawan hawan keke | Zazzabi | Lokacin amsawa | |
| 1 | 1 | 50 ℃ | 10 min | |
| 2 | 1 | 95 ℃ | 30 seconds | |
| 3 | 45 | 95 ℃ | 5 dakika | |
| 60 ℃ | 30 seconds | Tarin Fluorescence | ||
3) Saitunan ganowa:
An saita tashoshin ganowa zuwa FAM, VIC, ROX da CY5, daidai da ORF1ab, N gene, da E Gene, RNase P na ciki, bi da bi."Quencher Dye" da "Passive Reference" an saita zuwa "Babu" don kayan aikin ABI 7500.Saita Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa ) da Samfura (Ba a sani ba) a cikin abin da samfurori suka dace, kuma saita sunan samfurin a cikin "Sample Name".
Don X-POCH16, aiki da shirin sune kamar haka:
1) Bayan an gama gwajin kai, buɗe murfin kuma sanya bututun amsawar PCR a cikin wuraren da aka keɓe a cikin kayan aiki.
2) Fara da zabar "Exper."zaɓi.Zaɓi zaɓin "Dukkan" ko kuma da hannu zaɓi wurin amsawa a gefen hagu na allon.
3) Zaɓi zaɓin "LOAD";zaɓi shirin gwajin;danna "AIKATA" da "RUN".Shirin yana ɗaukar 30min42s don kammalawa.
An saita tashoshin ganowa na Default shirin zuwa FAM, VIC, ROX da CY5, daidai da ORF1ab, N gene, da E Gene, RNase P na ciki, bi da bi.
Ma'aunin zagayowar shirin Default sune kamar haka:
| Shirin | Adadin | Zazzabi | Lokacin amsawa |
| 1 | 1 | 50 ℃ | 2 min |
| 2 | 1 | 95 ℃ | 30 seconds |
| 3 | 41 | 95 ℃ | 2 dakika |
| 60 ℃ | dakika 13 | Fluorescence |
5. Saitin Ƙarfi
Dangane da hoton da aka bincika, daidaita ƙimar Fara, Ƙimar Ƙarshen Ƙimar Baseline da Ƙimar Ƙarshen (ƙimar farawa da ƙimar ƙarshe ana ba da shawarar saita su su zama 3 da 15 bi da bi, kuma ana daidaita lanƙwan haɓakar ƙarancin iko don zama lebur ko ƙasa da layin kofa), danna Analysis don samun bincike ta atomatik samfurin ƙimar Ct.Duba sakamakon a cikin Rahoto dubawa.
6. Standard Control Standard
Kowane iko na kit ɗin dole ne ya cika buƙatu masu zuwa tare da lanƙwan 'S', in ba haka ba gwajin ba ya aiki.
| Tashoshin ganowa | Sarrafa mara kyau | Kyakkyawan iko |
| FAM(ORF1ab) | Babu Ct | Ct≤38 |
| VIC(N) | Babu Ct | Ct≤38 |
| ROX(E) | Babu Ct | Ct≤38 |
| CY5(RP) | Babu Ct | Ct≤38 |
【Yanke Ƙimar】
Dangane da sakamakon samfuran swab na oropharyngeal 100 da samfuran sputum 100, kuma tare da hanyar lanƙwasa ROC, ƙimar Cut-off na OFR1ab, N genes E Gene na wannan kit shine Ct = 38
FAQ
A cikin wannan kit ɗin, firamare da bincike na fasaha na PCR mai kyalli na ainihin lokaci an ƙirƙira su zuwa takamaiman yankuna na ORF1ab, N da E na 2019-nCoV bi da bi.A lokacin haɓakawa na PCR, binciken yana ɗaure da samfuri, kuma ƙungiyar masu ba da rahoto ta 5'-ƙarshen binciken tana katsewa ta hanyar Taq enzyme (5'→3' ayyukan exonuclease), don haka yana motsawa daga ƙungiyar kashewa don samar da siginar kyalli. .Ana ƙididdige maɓallin ƙarawa na ainihin lokaci ta atomatik bisa siginar haske da aka gano, kuma ana ƙididdige ƙimar samfurin Ct.FAM, VIC da ROX fluorophores ana lakafta su zuwa ORF1ab gene, N gene da E bincike.Ta amfani da gwaji guda ɗaya, ana iya gano ƙimar ƙimar waɗannan kwayoyin halitta guda uku na 2019-nCoV a lokaci guda.
An ba da kit ɗin tare da kulawa na ciki wanda ke niyya ga halittar RNase P don saka idanu tarin samfuran asibiti, sarrafawa, cirewa da tsarin RT-PCR don guje wa sakamako mara kyau.Ana yiwa ikon cikin gida lakabi da ƙungiyar masu kyalli na CY5.
1. Yi nazari da fassara sakamakon gwajin lokacin da kayan aiki ya kasance na al'ada, da kuma kulawa mai kyau, kulawa mara kyau da sakamakon gwaji na ciki sun hadu da daidaitattun kulawa.
2. Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙarfafawa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwar Ƙaƙwalwa na Ƙaƙwalwar Ƙaƙwalwar Ƙaƙwalwar Ƙaƙƙarfan Ƙaƙwalwar Ƙarƙashin Ƙarƙashin Ƙarƙashin Ƙarƙa ) ya nuna na S-curve da Ct ≤ 38, fassarar sakamakon kwayoyin da aka yi niyya yana ƙarƙashin yanayi masu zuwa.
| Tashoshin ganowa | Fassarar sakamakon kwayoyin da aka yi niyya | ||
| FMA | VIC (N gene) | ROX (E gene) | |
| Ct≤38 | Ct≤38 | Ct≤38 | Tare da madaidaicin ƙara girman S, ƙimar Ct shine≤38, madaidaicin ƙwayar maƙasudi yana da inganci. |
| 38 Ct 40 | 38 Ct 40 | 38 Ct 40 | Tare da yanayin ƙara girman S, sake gwada madaidaicin maƙasudin samfurin samfurin. Idan ƙimar Ct＜ 40 tare da madaidaicin madaidaicin ƙara girman S, madaidaicin jigon manufa yana da inganci;idan darajar Ct≥40, madaidaicin kwayar halitta mara kyau |
| Ct≥40 | Ct≥40 | Ct≥40 | Madaidaicin kwayar halittar manufa mara kyau |
Fassarar sakamako na 2019-nCoV:
Bisa ga sakamakon ORF1ab, N gene da E gene, fassarar kamar haka:
1) Idan kwayoyin halitta guda biyu ko uku na kwayoyin halittar da aka gano suna da inganci, 2019-nCoV yana da inganci;
2) Idan DAYA ko BABU daga cikin kwayoyin halittar da aka gano suna da inganci, 2019-nCoV mara kyau.
Lura: Madaidaicin ƙarawa na ingantaccen samfurin yakamata ya kasance tare da madaidaicin S.Koyaya, idan maƙasudin maƙasudin ya yi yawa, ƙila ba za a iya haɓaka daidaitaccen kulawar cikin gida ba kuma ana iya tantance samfurin kai tsaye a matsayin tabbatacce.Idan kowane ɗayan kwayoyin halittar biyu da aka yi niyya ya sami Ct≤38, 2019-nCoV yana da inganci.Idan kowane ɗayan kwayoyin halittar biyu da aka yi niyya sun sami Ct≥40, 2019-nCoV mara kyau.Idan Ct≥40, ko nuna babu ƙima, fassarar sakamakon halittar da aka yi niyya mara kyau ne.
3. Idan duk darajar Ct na tashoshin FAM, VIC, ROX da Cy5 sun fi 38 ko kuma babu wata alamar ƙararrawar S.
1) Akwai / akwai abubuwa (s) a cikin samfurin da ke hana amsawar PCR.Ana bada shawara don tsoma samfurin don sake gwadawa.
2) Tsarin hakar nucleic acid ba shi da kyau, don haka ana ba da shawarar sake cirewa.
nucleic acid don sake gwadawa.
3) Wannan samfurin ba samfurin da ya dace ba ne a lokacin yin samfur, ko ƙasƙanta
a lokacin sufuri da ajiya.
Hanyoyi 2 ne da za mu iya gano wannan yanayin: NAAT da Antigen.
(Ya zo daga CDC na Los Angeles)
