I-Monkeypox Virus (MPV) i-Nucleic Acid Detection Kit

Ikhithi iqulethe ii-reagents zokuqhuba iimvavanyo ezingama-48 okanye ulawulo lomgangatho, kubandakanya la macandelo alandelayo:

I-Reagent A

I-ReagentB

Qaphela: Amalungu amanani eebhetshi ahlukeneyo akanakusetyenziswa ngokutshintshanayo

【Iimeko zoGcino kunye noBomi beShelf】

I-1.Reagent A / B inokugcinwa kwi-2-30 ° C, kwaye ubomi beshelufu ziinyanga ezili-10.

2.Nceda uvule ikhava yetyhubhu yovavanyo kuphela xa ulungele uvavanyo.

3.Musa ukusebenzisa iityhubhu zovavanyo ngaphaya komhla wokuphelelwa.

4.Musa ukusebenzisa ityhubhu yokubona evuzayo.

【Isixhobo Esisebenzayo】

Ifanelekile kwi-LC480 PCR inkqubo yokuhlalutya, i-Gentier 48E Automatic PCR system analysis, ABI7500 PCR system analysis.

【IiMfuno zeSampulu】

1.Iintlobo zeesampuli ezisebenzayo: iisampulu zomqala.

2.Isisombululo sesampulu:Emva kokuqinisekiswa, kuyacetyiswa ukuba kusetyenziswe i-saline eqhelekileyo okanye ityhubhu yogcino lweNtsholongwane eveliswe yi-Hangzhou Testsea biology ukuqokelela isampulu.

umqala:sula iitoni ze-pharyngeal zamazwe amabini kunye nodonga lwangasemva lwe-pharyngeal kunye ne-sposal sterile sampling swab, cwilisa i-swab kwi-tube equlethe isisombululo se-sampling ye-3mL, ulahle umsila, kwaye uqinise isigqubuthelo sombhobho.

3. Ukugcinwa kwesampuli kunye nokuhanjiswa:Iisampulu eziza kuvavanywa kufuneka zivavanywe ngokukhawuleza.Ubushushu bokuthutha kufuneka bugcinwe kwi-2 ~ 8 ℃. Iisampuli ezinokuvavanywa kwiiyure ezingama-24 zingagcinwa kwi-2 ℃ ~ 8 ℃ kwaye ukuba iisampuli azikwazi ukuvavanywa kwiiyure ezingama-24, kufuneka zigcinwe ngaphantsi okanye zilingana. ukuya ku -70 ℃ (ukuba akukho meko yokugcina -70 ℃, inokugcinwa ku -20 ℃ okwethutyana), kunqande ukuphinda

ukukhenkceza nokunyibilika.

I-4.Ukuqokelela isampula efanelekileyo, ukugcinwa, kunye nokuthutha kubaluleke kakhulu ekusebenzeni kwale mveliso.

【Indlela yoVavanyo】

1.Ukwenziwa kwesampuli kunye nokudibanisa isampuli

1.1 Ukwenziwa kwesampuli

Emva kokuxuba isisombululo sesampulu esingentla kunye neesampulu, thatha i-30μL yesampuli kwi-DNA yokukhupha i-reagent tube kwaye uyixube ngokulinganayo.

1.2 Iyalayisha

Thatha i-20μL ye-reagent yokubuyisela kwaye uyifake kwi-reagent yokufumanisa i-MPV, yongeza i-5μL yesampuli eqhutywe ngasentla (Ulawulo olulungileyo kunye nolawulo olubi luya kucutshungulwa ngokuhambelana neesampuli), vala i-tube cap, i-centrifuge kwi-2000rpm ye-10. imizuzwana.

2. Ukwandiswa kwePCR

2.1 Layisha ipleyiti yePCR elungisiweyo/iityhubhu kwisixhobo sePCR sefluorescence, Ulawulo olubi kunye nolawulo olulungileyo luya kumiselwa kuvavanyo ngalunye.

2.2 Ukusetwa kwetshaneli yeFluorescent:

1) Khetha itshaneli ye-FAM yokuchongwa kweMPV;

2) Khetha itshaneli ye-HEX/VIC yolawulo lwangaphakathi lofuzo;

3.Uhlalutyo lweziphumo

Seta umgca wesiseko ngentla kweyona ndawo iphezulu yegophe le-fluorescent yolawulo olungalunganga.

4.Ulawulo lomgangatho

4.1 Ulawulo olubi: Akukho xabiso le-Ct lifunyenwe kwi-FAM, i-HEX / VIC channel, okanye i-Ct＞40;

4.2 Ulawulo oluhle: Kwi-FAM, itshaneli ye-HEX/VIC, Ct≤40;

4.3 Ezi mfuno zingentla kufuneka zaneliswe kuvavanyo olufanayo, kungenjalo iziphumo zovavanyo azivumelekanga kwaye uvavanyo kufuneka luphindwe.

【Sika ixabiso】

Isampulu ithathwa njengento entle xa: Ulandelelwano ekujoliswe kulo Ct≤40, Igene yolawulo lwangaphakathi Ct≤40.

【Ukutolikwa kweziphumo】

Nje ukuba ulawulo lomgangatho lugqithiselwe, abasebenzisi kufuneka bajonge ukuba ngaba kukho ijika lokukhulisa kwisampulu nganye kwitshaneli ye-HEX/VIC, ukuba kukho kunye ne-Ct≤40, ibonise ukuba ijini yolawulo lwangaphakathi yandisiwe ngempumelelo kwaye olu vavanyo luchanekileyo.Abasebenzisi banokuqhubeka nohlalutyo olulandelayo:

3.Kwiisampulu ezinokwandiswa kwejini yolawulo lwangaphakathi ayiphumelelanga (HEX/VIC

itshaneli, Ct＞40, okanye akukho curve yokukhulisa), umthamo weViral ophantsi okanye ubukho be-PCR inhibitor bunokuba sisizathu sokungaphumeleli, uviwo kufuneka luphindwe kwingqokelela yomfanekiso;

I-4.Iisampulu ezintle kunye nentsholongwane ekhulisiwe, iziphumo zolawulo lwangaphakathi azichaphazeli;

Kwiisampuli ezifunyaniswe zi-negative, ulawulo lwangaphakathi kufuneka luvavanywe ukuba unayo, kungenjalo iziphumo zizonke azikho mthethweni kwaye uvavanyo kufuneka luphindwe, kuqalwa kwinyathelo lokuqokelelwa kwesampula.