I-Monkeypox Virus (MPV) Nucleic Acid Detection Kit

Ikhithi iqukethe ama-reagents okucubungula izivivinyo ezingama-48 noma ukulawulwa kwekhwalithi, okuhlanganisa izingxenye ezilandelayo:

I-Reagent A

I-ReagentB

Qaphela: Izingxenye zezinombolo zeqoqo ezahlukene azikwazi ukusetshenziswa ngokushintshana

【Izimo Zesitoreji Kanye Nempilo Yeshelufu】

1.I-Reagent A/B ingagcinwa ku-2-30°C, kanti isikhathi seshelufu siyizinyanga eziyi-10.

2.Sicela uvule ikhava yeshubhu yokuhlola kuphela uma usulungele ukuhlolwa.

3.Ungasebenzisi amashubhu okuhlola ngaphezu kosuku lokuphelelwa yisikhathi.

4.Ungasebenzisi ishubhu lokubona elivuzayo.

【Ithuluzi Elisebenzayo】

Ifanele isistimu yokuhlaziya ye-LC480 PCR, i-Gentier 48E Automatic PCR system analysis, ABI7500 PCR system analysis.

【Izidingo Zesampula】

1.Izinhlobo zesampula ezisebenzayo: amasampula e-throat swabs.

2.Isixazululo sesampula:Ngemva kokuqinisekisa, kunconywa ukusebenzisa usawoti ovamile noma ishubhu lokulondoloza igciwane elikhiqizwe yi-Hangzhou Testsea biology ukuze kuqoqwe isampula.

umqala:sula amathoni epharyngeal amabili kanye nodonga lwangemuva lwepharyngeal ngeswabhu yesampula eyinyumba elahlwayo, cwilisa i-swab eshubhuni eliqukethe isisombululo sesampula esingu-3mL, ulahle umsila, bese uqinisa ikhava yeshubhu.

3. Ukugcinwa kwesampula nokulethwa:Amasampula azohlolwa kufanele ahlolwe ngokushesha ngangokunokwenzeka.Izinga lokushisa lokuthutha kufanele ligcinwe ku-2 ~ 8 ℃. Amasampula angahlolwa kungakapheli amahora angu-24 angagcinwa ku-2 ℃ ~ 8 ℃ futhi uma amasampula engakwazi ukuhlolwa kungakapheli amahora angu-24, kufanele agcinwe ngaphansi noma alinganayo. kuya ku -70 ℃ (uma singekho isimo sokugcina sika -70 ℃, singagcinwa ku -20 ℃ okwesikhashana), gwema ukuphindaphinda

ukuqhwaza nokuncibilika.

I-4.Iqoqo lesampula elifanelekile, ukugcinwa, nokuthutha kubalulekile ekusebenzeni kwalo mkhiqizo.

【Indlela Yokuhlola】

1.Ukucubungula isampula kanye nokwengezwa kwesampula

1.1 Ukucubungula isampula

Ngemva kokuxuba isixazululo sesampula esingenhla namasampula, thatha u-30μL wesampula uwufake eshubhuni ye-DNA ekhipha isampula bese uyixuba ngokulinganayo.

1.2 Iyalayisha

Thatha u-20μL we-reagent yokuhlanganisa futhi uyengeze ku-reagent yokuthola i-MPV, engeza u-5μL wesampula ecutshunguliwe ngenhla (Ukulawula okuhle nokulawula okungalungile kuzocutshungulwa ngokuhambisana namasampuli), vala i-tube cap, uyibeke phakathi ku-2000rpm ngo-10. imizuzwana.

2. Ukukhulisa i-PCR

2.1 Layisha ipuleti/amashubhu e-PCR alungisiwe ensimbini ye-fluorescence PCR, Ukulawula okungekuhle kanye nokulawula okuhle kuzosethelwa ukuhlolwa ngakunye.

2.2 Ukulungiselelwa kwesiteshi seFluorescent:

1) Khetha isiteshi se-FAM ukuze uthole i-MPV;

2) Khetha isiteshi se-HEX/VIC ukuze uthole ukutholwa kofuzo lwangaphakathi;

3.Ukuhlaziywa kwemiphumela

Setha umugqa wesisekelo ngaphezu kwephoyinti eliphakeme kakhulu lejika le-fluorescent lesilawuli esinegethivu.

4.Ukulawulwa kwekhwalithi

4.1 Ukulawula okungekuhle: Alikho inani le-Ct elitholwe ku-FAM, i-HEX/VIC isiteshi, noma i-Ct＞40;

4.2 Ukulawula okuhle: Ku-FAM, HEX/VIC channel, Ct≤40;

4.3 Lezi zidingo ezingenhla kufanele zaneliswe esivivinyweni esifanayo, ngaphandle kwalokho imiphumela yokuhlolwa ayivumelekile futhi isilingo sidinga ukuphindwa.

【Sika inani】

Isampula libhekwa njengelihle uma: Ukulandelana kwethagethi Ct≤40, Igene yokulawula yangaphakathi Ct≤40.

【Ukuchazwa kwemiphumela】

Uma isilawuli sekhwalithi sesidlulisiwe, abasebenzisi kufanele bahlole ukuthi ingabe ikhona yini ijika lokukhulisa isampula ngalinye esiteshini se-HEX/VIC, uma likhona futhi nge-Ct≤40, ibonise uhlobo lofuzo lokulawula lwangaphakathi lukhuliswe ngempumelelo futhi lokhu kuhlola okuthile kuvumelekile.Abasebenzisi bangaqhubekela ekuhlaziyweni okulandelayo:

3.Kumasampuli anokukhulisa ufuzo lokulawula kwangaphakathi kwehlulekile (HEX/VIC

isiteshi, i-Ct＞40, noma alikho ijika lokukhulisa), umthamo weViral ophansi noma ukuba khona kwe-PCR inhibitor kungaba isizathu sokwehluleka, ukuhlolwa kufanele kuphindwe kusukela eqoqweni lesampula;

4.Kumasampula amahle kanye negciwane elikhulisiwe, imiphumela yokulawula kwangaphakathi ayithinti;

Kumasampuli ahlolwe ukuthi akanayo, isilawuli sangaphakathi sidinga ukuhlolwa ukuthi unayo ngaphandle kwalokho umphumela usuwonke awuvumelekile futhi ukuhlolwa kudinga ukuphindwa, kusukela esinyathelweni sokuqoqwa kwesifanekiso.