Virusi ya Monkeypox (MPV) Nucleic Acide Detection Kit

Igikoresho kirimo reagent zo gutunganya ibizamini 48 cyangwa kugenzura ubuziranenge, harimo ibice bikurikira:

Reagent A.

ReagentB

Icyitonderwa: Ibice byimibare itandukanye ntishobora gukoreshwa muburyo bumwe

【Imiterere yo kubika hamwe nubuzima bwa Shelf】

1.Ragent A / B irashobora kubikwa kuri 2-30 ° C, kandi igihe cyo kubaho ni amezi 10.

2. Nyamuneka fungura igifuniko cya test tube gusa mugihe witeguye gukora ikizamini.

3.Ntukoreshe ibizamini byo kwipimisha kurenza itariki izarangiriraho.

4.Ntugakoreshe umuyoboro wamenyekanye.

【Igikoresho gikoreshwa】

Bikwiranye na LC480 sisitemu yo gusesengura PCR, Gentier 48E Sisitemu yo gusesengura PCR yikora, sisitemu yo gusesengura ABI7500 PCR.

【Icyitegererezo】

1.Ubwoko bw'icyitegererezo bukoreshwa: umuhogo wo mu muhogo.

2.Icyitegererezo:Nyuma yo kugenzurwa, birasabwa gukoresha umuyoboro usanzwe wa saline cyangwa Virusi yo kubika virusi yakozwe na biologiya ya Hangzhou Testsea kugirango ikusanyirizwe hamwe.

umuhogo:ohanagura toni ya faryngeal byombi hamwe nurukuta rwinyuma rwa faryngeal hamwe na swabable sterile sampling swab, wibiza swab mu muyoboro urimo igisubizo cya 3mL, guta umurizo, no gukomera umupfundikizo.

3.Urugero rwo kubika no gutanga:Ingero zigomba gupimwa zigomba gupimwa vuba bishoboka.Ubushyuhe bwo gutwara abantu bugomba kubikwa kuri 2 ~ 8 ℃ .Urugero rushobora kugeragezwa mu masaha 24 rushobora kubikwa kuri 2 ℃ ~ 8 ℃ kandi niba ingero zidashobora gupimwa mu masaha 24, zigomba kubikwa munsi cyangwa zingana. kugeza kuri -70 ℃ (niba nta bubiko bwa -70 ℃, burashobora kubikwa kuri -20 ℃ by'agateganyo), irinde gusubiramo

gukonjesha no gukonja.

4.Icyegeranyo gikwiye cyo gukusanya, kubika, no gutwara abantu ni ngombwa mu mikorere yiki gicuruzwa.

【Uburyo bwo Kwipimisha】

1.Urugero rwo gutunganya no kongeramo icyitegererezo

1.1 Gutunganya icyitegererezo

Nyuma yo kuvanga igisubizo cyavuzwe haruguru hamwe nicyitegererezo, fata 30μL yicyitegererezo muri ADN irekura reagent hanyuma ubivange neza.

1.2 Kuremera

Fata 20μL ya reagent yo kwiyubaka hanyuma uyongere kuri reagent ya MPV, ongeramo 5μL yicyitegererezo cyatunganijwe haruguru (Igenzura ryiza hamwe nigenzura ribi bizakorwa bigereranywa nicyitegererezo), upfundikire igituba, centrifuge kuri 2000rpm kuri 10 amasegonda.

2. Kwiyongera kwa PCR

2.1 Shira plaque ya PCR yateguwe kubikoresho bya fluorescence PCR, kugenzura nabi no kugenzura neza bizashyirwaho kuri buri kizamini.

2.2 Gushiraho umuyoboro wa Fluorescent ：

1) Hitamo umuyoboro wa FAM kugirango umenye MPV ；

2) Hitamo umuyoboro wa HEX / VIC kugirango ugenzure imbere gene ；

3.Isesengura ryibisubizo

Shyira umurongo shingiro hejuru yumurongo wo hejuru wigenzura ribi rya fluorescent curve.

4. Kugenzura ubuziranenge

4.1 Igenzura ribi ： Nta Ct agaciro yagaragaye muri FAM channel HEX / VIC, cyangwa Ct ＞ 40 ；

4.2 Igenzura ryiza ： Muri FAM channel HEX / VIC umuyoboro, Ct≤40 ；

4.3 Ibisabwa haruguru bigomba kuba byujujwe mubigeragezo bimwe, bitabaye ibyo ibisubizo byikizamini bikaba bitemewe kandi ubushakashatsi bugomba gusubirwamo.

【Gabanya agaciro】

Icyitegererezo gifatwa nkicyiza iyo: Intego ikurikirana Ct≤40, Imbere yo kugenzura imbere Ct≤40.

【Ibisobanuro】

Igenzura rimaze gutambuka, abayikoresha bagomba kugenzura niba hari umurongo wa amplification umurongo kuri buri cyitegererezo mu muyoboro wa HEX / VIC, niba uhari hamwe na Ct≤40, byerekanaga ko gen igenzura imbere yongerewe neza kandi iki kizamini gifite ishingiro.Abakoresha barashobora gukomeza gukurikirana isesengura:

3.Kurugero hamwe na amplification ya gen igenzura imbere byananiranye (HEX / VIC

umuyoboro, Ct ＞ 40, cyangwa nta amplification curve), umutwaro muke wa virusi cyangwa kubaho kwa PCR inhibitor bishobora kuba impamvu yo gutsindwa, ikizamini kigomba gusubirwamo uhereye kubikusanyamakuru;

4.Kurugero rwiza na virusi yumuco, ibisubizo byo kugenzura imbere ntabwo bigira ingaruka;

Kubitegererezo byageragejwe nabi, igenzura ryimbere rigomba gupimwa neza naho ubundi ibisubizo muri rusange ntibyemewe kandi ikizamini kigomba gusubirwamo, guhera kumurongo wo gukusanya icyitegererezo.