I-Reagent yokukhishwa kwesampula
Isampula yokukhishwa kwe-Release Reagent eyezimo zokuxilonga ze-molecular POCT.Kulezi zinhlelo ezimbili ze-LAMP yokukhulisa i-amplification eqondile kanye ne-PCR yokukhulisa i-amplification eqondile, asikho isidingo sokukhishwa kwe-nucleic acid.I-lysate engahluziwe yesampula ingakhuliswa ngokuqondile, isakhi sofuzo esiqondiwe singatholwa ngokunembile, isikhathi sokuthola isampula singancishiswa ngokuqhubekayo, esifana ngokuphelele nezidingo zohlelo lokusebenza lwe-POCT yamangqamuzana.Ifanele ama-nasal swabs, ama-throat swabs nezinye izinhlobo zesampula.Amasampuli acutshunguliwe angasetshenziswa ngokuqondile ekutholeni i-PCR yesikhathi sangempela se-fluorescence quantitative noma i-LAMP, futhi imiphumela efanayo nezindlela ezivamile zokukhipha ingafinyelelwa ngaphandle kokusebenza okuyinkimbinkimbi kokukhipha i-nucleic acid.
Izimo Zokugcina
Ezokuthutha kanye nesitolo ekamelweni lokushisa.
Ikhwalithi yokulawula
Ukutholwa okusebenzayo - i-qPCR yobuningi: Isistimu ye-Release Release Reagent engu-800μl iye yandiswa
enamakhophi ayi-1000 i-Novel Pseudovirus, isampula elilodwa le-swab yamakhala, okuholela amajika afanayo okukhulisa kanyeΔCt amanani angaphakathi kuka-± 0.5 Ct.
Inqubo Yokuhlolares
1. Thatha i-800 μl Release Reagent yesampula bese usakaza isisombululo se-lysis ku-1.5 mL yesampula yeshubhu.
2. Thatha i-swab yamakhala noma i-swab yomphimbo nge-swab;Inqubo yesampula ye-swab yamakhala: thatha i-swab oyinyumba uyifake emakhaleni, kancane kancane uqhubekele ku-1.5 cm ukushona, ujikeleze ngobumnene izikhathi ezingu-4 ngokumelene ne-mucosa yamakhala imizuzwana engaphezu kwe-15. , bese uphinda ukuhlinzwa okufanayo komunye umgodi wekhala nge-swab efanayo.Inqubo yokusampula yomphimbo wesampula: thatha i-swab oyinyumba futhi ngobumnene, usule ngokushesha amathoni e-pharyngeal kanye nodonga lwangemuva lwe-pharyngeal izikhathi ezingu-3.
3.Faka i-swab ngokushesha kushubhu yesampula.Ikhanda le-swab kufanele lijikelezwe futhi lixutshwe kusixazululo sesitoreji okungenani imizuzwana engama-30 ukuze kuqinisekiswe ukuthi isampula likhishwe ngokugcwele kushubhu lesampula.
4. Ukufukamela ekamelweni lokushisa (20~ 25℃) i-1min, ukulungiswa kwe-lysis buffer kuqediwe.
5. Kokubili isistimu ye-25μl ye-RT-PCR ne-RT-LAMP yayihambisana nenani elingu-10μl lokwengezwa kwesifanekiso sokuhlolwa kokutholwa.
Amanothi
1. Inani elincane lesampula le-lysate eqondile elihambisana ne-swab eyodwa lingalungiswa libe ngu-400μl, elingalungiswa ngokuvumelana nezidingo zokuhlola.
2. Uma isampula isicutshungulwe yi-reagent yokukhishwa kwesampula, kunconywa ukwenza isigaba esilandelayo sokuhlola ngokushesha okukhulu, isikhathi sokulinda sesikhawu singaphansi kwehora elingu-1.
3. I-pH yesampula ye-lysate ine-asidi, futhi isistimu yokuthola idinga ukuba nesibhafa esithile.Ilungele ukutholwa okuningi kwe-PCR, RT-PCR, kanye ne-LAMP fluorescence ene-pH buffer, kodwa ayifanele ukutholwa kombala we-LAMP ngaphandle kwebhafa.
