Monkeypox Virus Igihe Cyukuri PCR Kit
Ibisobanuro birambuye
Gukoresha
Ikoreshwa mugutahura virusi ya Monkeypox muri serumu yumuntu cyangwa lesion exudate sample ukoresheje sisitemu nyayo PCR.
Ihame ry'ikizamini
Ibicuruzwa ni fluorescent probe ishingiye kuri Taqman® sisitemu nyayo ya PCR.Primers na probe zihariye zagenewe kumenya gene ya F3L ya virusi ya Monkeypox.Igenzura ryimbere ryibasira gene yabantu ikurikirana ikusanyamakuru ryikitegererezo, uburyo bwo gukora icyitegererezo hamwe nigihe nyacyo PCR kugirango wirinde ibisubizo bibi-bibi.Igikoresho ni sisitemu yuzuye ya lyophilized sisitemu, ikubiyemo ibikoresho bikenerwa mugushakisha virusi ya Monkeypox: enzyme ya nucleic aside amplification enzyme, enzyme ya UDG, reaction reaction, primer na probe.
Ibirimo
Ibice byatanzwe biri kurutonde.
| Ibigize | Amapaki | Ibikoresho |
| Virusi ya MonkeypoxLyophilized Premix | 8 kwiyambura imiyoboro ya PCRAmapaki 6 | Primers, probe, DNTP / dUTP Ivanga, Mg2 +, Taq DNA polymerase, UDG Enzyme |
| MPV Igenzura ryiza | 400 μL × 1 umuyoboro | Urutonde rwa ADN ikubiyemo intego ya gen |
| Kugenzura nabi MPV | 400 μL × 1 umuyoboro | Urutonde rwa ADN rurimo igice cya gen |
| Gukemura igisubizo | 1 mL × 1 umuyoboro | Stabilizer |
| Icyemezo cyo guhuza | Igice 1 | / |
Ibikorwa
1. IcyitegererezoIcyegeranyo:Ibigereranirizo bigomba gukusanyirizwa mu tubari sterile ukurikije
hamwe nibisanzwe bya tekiniki.
2. Gutegura Reagent (Agace ko Gutegura)
Kuramo ibice bigize ibikoresho, uburinganire mubushyuhe bwicyumba kugirango ukoreshwe neza.
3. Gutunganya Ibigereranyo (Agace gatunganyirizwamo)
3.1 Gukuramo aside nucleique
Birasabwa gufata urugero rwa 200μL rwamazi, kugenzura neza no kugenzura nabi gukuramo aside nucleic, ukurikije ibisabwa hamwe nuburyo bukoreshwa mubikoresho byo gukuramo ADN virusi.
3.2 Ifu ya Liyofilize ishonga no kongeramo inyandikorugero
Tegura Monkeypox Virus Lyophilized premix ukurikije umubare wintangarugero.Icyitegererezo kimwe gikenera umuyoboro umwe wa PCR urimo ifu ya Lyophilized.Kugenzura nabi & kugenzura neza bigomba gufatwa nkintangarugero ebyiri.
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.niba ukoresha BTK-8 mugushakisha, noneho ukeneye gukora ibikorwa bikurikira: kwimura 10 μL amazi ava mumiyoboro ya PCR kuri chip reaction ya BTK-8.Umuyoboro umwe wa PCR uhuye n'iriba rimwe kuri chip.Mugihe cyo gukora imiyoboro, menya neza ko pipeti ihagaze dogere 90.Inama ya bariyeri ya aerosol igomba gushyirwa hagati y iriba n'imbaraga ziciriritse hanyuma ukareka gusunika pipeti iyo igeze kubikoresho byambere (kugirango wirinde ibibyimba).Amariba amaze kuzura, fata reaction ya chip membrane kugirango utwikire amariba yose hanyuma chip ihindurwe ahantu hagaragara.
4. Kwiyongera kwa PCR (Agace ko gutahura)
4.1 Shira PCR tubes / reaction chip muri reaction hanyuma ushireho amazina ya buri reaction neza murutonde rukwiranye.
4.2 Igenamiterere rya fluorescence: (1) virusi ya Monkeypox (FAM);(2) Igenzura ryimbere (CY5).
4.3 Koresha protocole ikurikira
Porotokole ya ABI7500, Bio-Rad CFX96, SLAN-96S, QuantStudio:
| Intambwe | Ubushyuhe | Igihe | Amagare | |
| 1 | Mbere yo gutandukana | 95 ℃ | Iminota 2 | 1 |
| 2 | Gutandukana | 95 ℃ | 10 s | 45 |
| Annealing, kwaguka, kugura fluorescence | 60 ℃ | 30 s | ||
Porotokole ya BTK-8:
| Intambwe | Ubushyuhe | Igihe | Amagare | |
| 1 | Mbere yo gutandukana | 95 ℃ | Iminota 2 | 1 |
| 2 | Gutandukana | 95 ℃ | 5 s | 45 |
| Annealing, kwaguka, kugura fluorescence | 60 ℃ | 14 s | ||
5. Isesengura ry'ibisubizo (nyamuneka reba Igitabo gikoresha Igitabo）
Nyuma yo kubyitwaramo, ibisubizo bizabikwa mu buryo bwikora.Kanda "Gusesengura" kugirango usesengure, kandi igikoresho kizahita gisobanura indangagaciro za Ct ya buri sample mubisubizo inkingi.Ibisubizo bibi kandi byiza byo kugenzura bigomba guhura nibi "6. Kugenzura ubuziranenge".
6. Kugenzura ubuziranenge
6.1 Igenzura ribi: Nta Ct cyangwa Ct> 40 mumuyoboro wa FAM, Ct≤40 mumuyoboro wa CY5 hamwe na amplification isanzwe.
6.2.
6.3 Ibisubizo biremewe niba ibisabwa byose byavuzwe haruguru byujujwe.Bitabaye ibyo, ibisubizo ntabwo byemewe.
Ibisobanuro
Ibisubizo bikurikira birashoboka:
| Ct agaciro k'umuyoboro wa FAM | Ct agaciro k'umuyoboro CY5 | Gusobanura | |
| 1# | Nta Ct cyangwa Ct> 40 | ≤40 | Virusi ya Monkeypox mbi |
| 2# | ≤40 | Ibisubizo byose | Virusi ya Monkeypox nziza |
| 3# | 40 ～ 45 | ≤40 | Ongera ugerageze;niba bikiri 40 ~ 45, raporo nka 1 # |
| 4# | Nta Ct cyangwa Ct> 40 | Nta Ct cyangwa Ct> 40 | Ntibyemewe |
ICYITONDERWA: Niba ibisubizo bitemewe bibaye, icyitegererezo gikeneye gukusanywa no kongera kugeragezwa.
Tegeka amakuru
| izina RY'IGICURUZWA | Injangwe.Oya | Ingano | Ingero | Ubuzima bwa Shelf | Trans.& Sto.Ubushuhe. |
| Monkeypox Virus Igihe Cyukuri PCR Kit | B001P-01 | Ibizamini 48 | Serumu / Lesion Exudate | Amezi 12 | -25 ～ -15 ℃ |
