Ikhithi yokuhlola ngokushesha iMonkeypox Virus 2022 MPV Nucleic Acid Detection Kit PCR Test nge-CE Euro Certification

[ INTRODUCTION ]
*Ikhithi isetshenziselwa ukutholwa kwekhwalithi ye-in vitro yezigameko ezisolwayo ze-Monkeypox Virus (MPV), amacala ahlanganisiwe nezinye izimo *ezidinga ukuhlolelwa ukutheleleka ngeMonkeypox Virus.
*Ikhithi isetshenziselwa ukuhlonza isakhi sofuzo se-f3L se-MPV kuma-swabs omphimbo namasampula e-nasal swab.
*Imiphumela yokuhlolwa yale khithi ingeyenkomba yomtholampilo kuphela futhi akufanele isetshenziswe njengeyona ndlela yodwa yokuxilongwa komtholampilo. Kunconywa ukwenza ukuhlaziywa okuphelele kwesimo ngokusekelwe ekubonakalisweni komtholampilo kwesiguli kanye nezinye izivivinyo zaselabhorethri. 

[ ISICI SOMKHIQIZO ]

[ Principle ]
Le khithi ithatha ukulandelana okuqondile okulondoloziwe kofuzo lwe-MPV f3L njengendawo eqondiwe. Ubuchwepheshe besikhathi sangempela be-fluorescence quantitative PCR kanye nobuchwepheshe bokukhululwa okusheshayo be-nucleic acid busetshenziselwa ukuqapha i-viral nucleic acid ngokushintsha isignali ye-fluorescence yemikhiqizo yokukhulisa umsindo. Isistimu yokuthola ihlanganisa ukulawula kwekhwalithi yangaphakathi, esetshenziselwa ukuqapha ukuthi ingabe akhona yini ama-PCR inhibitor kumasampuli noma ukuthi amaseli kumasampuli athathiwe, okungavimbela ngempumelelo isimo esingesihle esingelona iqiniso.

[IMINININGWANE EMAKHULU]

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

I-Reagent A

Ukuphendula B

Qaphela: Izingxenye zezinombolo zeqoqo ezahlukene azikwazi ukusetshenziswa ngokushintshana

[ Izimo Zesitoreji Nempilo Yeshelufu ] 
1.I-Reagent A/B ingagcinwa ku-2-30°C, kanti impilo yeshelufu yizinyanga eziyi-10.
2.Sicela uvule ikhava yeshubhu yokuhlola kuphela uma usulungele ukuhlolwa.
3.Do not use test tubes beyond the expiration date.
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: throat swabs samples.
2.Isixazululo sesampula: Ngemva kokuqinisekisa, kunconywa ukusebenzisa usawoti ovamile noma ishubhu lokulondoloza igciwane elikhiqizwe yi-Hangzhou Testsea biology ukuze kuqoqwe isampula.
umqala: sula amathoni wepharyngeal omabili kanye nodonga lwangemuva lwepharyngeal ngeswabhu yesampula eyinyumba elahlwayo, cwilisa i-swab eshubhuni eliqukethe isisombululo sesampula esingu-3mL, ulahle umsila, bese uqinisa ikhava yeshubhu.
3.Isampula yokugcina nokulethwa: Amasampula azohlolwa kufanele ahlolwe ngokushesha ngangokunokwenzeka. Izinga lokushisa lokuthutha kufanele ligcinwe ku-2 ~ 8ºC. Amasampula angahlolwa phakathi namahora angu-24 angagcinwa ku-2ºC~8ºC futhi uma amasampula engakwazi ukuhlolwa phakathi namahora angu-24, kufanele agcinwe ngaphansi noma alingane no -70ºC. (uma singekho isimo sokugcina esingu -70ºC, singagcinwa ku -20ºC okwesikhashana), gwema ukuphindaphinda
ukuqhwaza nokuncibilika.
I-4.Iqoqo lesampula elifanelekile, ukugcinwa, nokuthutha kubalulekile ekusebenzeni kwalo mkhiqizo.
[Testing Method ]
1.Isampula yokucubungula kanye nokwengezwa kwesampula
1.1 Ukucutshungulwa kwesampula
Ngemva kokuxuba isixazululo sesampula esingenhla namasampuli, thatha u-30μL wesampula ulifake eshubhu le-reagent ekhipha i-DNA bese uyixuba ngokulinganayo.
1.2 Iyalayisha
Take 20μL of the reconstitution reagent and add it to the MPV detection reagent, add 5μL of the above processed sample (The positive control and negative control shall be processed in parallel with the samples), cover the tube cap, centrifuge it at 2000rpm for 10 seconds.
2. Ukukhulisa i-PCR
2.1 Load the prepared PCR plate/tubes to the fluorescence PCR instrument, Negative control and positive control shall be set for each test.
2.2 Ukulungiselelwa kwesiteshi se-Fluorescent:
1)Khetha isiteshi se-FAM ukuze uthole i-MPV;
2)Khetha isiteshi se-HEX/VIC ukuze uthole ukulawula kwangaphakathi kofuzo;
3.Ukuhlaziywa kwemiphumela
Setha umugqa wesisekelo ngaphezu kwephoyinti eliphakeme kakhulu lejika le-fluorescent lesilawuli esinegethivu.
4.Ukulawulwa kwekhwalithi
4.1 Ukulawula okungalungile: Alikho inani le-Ct elitholwe kusiteshi se-FAM, HEX/VIC, noma i-Ct>40;
4.2 Positive control:In FAM,HEX/VIC channel, Ct≤40;
4.3 Lezi zidingo ezingenhla kufanele zaneliswe esivivinyweni esifanayo, ngaphandle kwalokho imiphumela yokuhlola ayivumelekile futhi isilingo sidinga ukuphindwa.
[ Sika inani ]
A sample is considered as positive when: Target sequence Ct≤40, The internal control gene Ct≤40.
[ Ukutolikwa kwemiphumela ]
Uma isilawuli sekhwalithi sesidlulile, abasebenzisi kufanele bahlole ukuthi ingabe ikhona yini ijika lokukhulisa isampula ngayinye esiteshini se-HEX/VIC, uma likhona futhi nge-Ct≤40, ibonise ukuthi uhlobo lofuzo lokulawula lwangaphakathi lukhuliswe ngempumelelo futhi lokhu kuhlola okuthile kuvumelekile. Abasebenzisi bangaqhubekela ekuhlaziyweni okulandelayo:
3.For samples with the amplification of internal control gene failed (HEX/VIC
isiteshi, i-Ct>40, noma alikho ijika lokukhulisa), umthamo wegciwane egazini 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 kudingeka kuphindwe, kusukela esinyathelweni sokuqoqwa kwesifanekiso.

[ Ukupakisha nokuhamba ]

[ Mayelana NATHI ]