Monkeypox Virus (MPV) Nucleic Acid Detection Kit

Kit e na le li-reagents bakeng sa ho sebetsana le liteko tse 48 kapa taolo ea boleng, ho kenyelletsa le likarolo tse latelang:

Reagent A

ReagentB

Tlhokomeliso: Likarolo tsa linomoro tse fapaneng tsa batch li ke ke tsa sebelisoa ka ho fapanyetsana

【Maemo a polokelo le Bophelo ba Shelf】

1.Reagent A / B e ka bolokoa ho 2-30 ° C, 'me bophelo ba sethala ke likhoeli tse 10.

2.Ka kopo bula sekoahelo sa tube ea teko feela ha u se u itokiselitse tlhahlobo.

3.U se ke ua sebelisa li-tubes tsa teko ho feta letsatsi la ho fela.

4.U se ke ua sebelisa tube ea ho lemoha e lutlang.

【Seletsa se Sebetsang】

E loketse bakeng sa Sistimi ea tlhahlobo ea LC480 PCR, sistimi ea tlhahlobo ea Gentier 48E Automatic PCR, sistimi ea tlhahlobo ea ABI7500 PCR.

【Litlhoko tsa Mohlala】

1.Mefuta ea mehlala e sebetsang: lisampole tsa 'metso.

2.Sampling tharollo:Kamora ho netefatsoa, ​​ho khothaletsoa ho sebelisa tšupa e tloaelehileng ea letsoai kapa ea ho boloka kokoana-hloko e hlahisoang ke Hangzhou Testsea biology bakeng sa ho bokella sampole.

mometso:Hlakola lithane tse peli tsa pharyngeal le lebota le ka morao la pharyngeal ka swab ea sampole e hloekisitsoeng, kenya sesepa ka har'a tube e nang le 3mL sampling solution, u lahle mohatla, 'me u tiise sekoaelo sa tube.

3.Sample polokelo le delibari:Mehlala e lokelang ho lekoa e lokela ho lekoa kapele kamoo ho ka khonehang.Mocheso oa lipalangoang o lokela ho bolokoa ho 2 ~ 8 ℃. Mehlala e ka hlahlojoang ka hare ho lihora tse 24 e ka bolokoa ho 2 ℃ ~ 8 ℃ 'me haeba lisampole li ke ke tsa lekoa ka hare ho lihora tse 24, li lokela ho bolokoa ka tlase ho kapa ho lekana. ho ea ho -70 ℃ (haeba ho se na boemo ba polokelo ea -70 ℃, e ka bolokoa ho -20 ℃ ka nakoana), qoba ho pheta-pheta

ho qhibidiha le ho qhibidiha.

4.Pokello e nepahetseng ea mohlala, polokelo, le lipalangoang ke tsa bohlokoa mosebetsing oa sehlahisoa sena.

【Mokhoa oa ho Leka】

1.Sample processing le ho eketsa mohlala

1.1 Mohlala oa ho sebetsa

Kamora ho kopanya tharollo ea sampole e kaholimo le lisampole, nka 30μL ea sampole ka har'a tube ea reagent ea DNA ebe u e kopanya ka ho lekana.

1.2 Ho kenya

Nka 20μL ea reagent ea reconstitution 'me u e kenye mochineng oa ho lemoha oa MPV, eketsa 5μL ea sampuli e entsoeng ka holimo (Taolo e ntle le taolo e mpe e tla sebetsoa ka ho tsamaisana le lisampole), koahela sekoahelo sa tube, se centrifuge ka 2000rpm bakeng sa 10. metsotsoana.

2. PCR amplification

2.1 Laola poleiti ea PCR e lokiselitsoeng ho sesebelisoa sa PCR sa fluorescence, Taolo e mpe le taolo e ntle e tla beoa bakeng sa tlhahlobo ka 'ngoe.

2.2 Tlhophiso ea kanale ea Fluorescent:

1) Khetha mocha oa FAM bakeng sa ho lemoha MPV;

2) Khetha mocha oa HEX / VIC bakeng sa ho lemoha liphatsa tsa lefutso tsa ka hare;

3.Tlhahlobo ea liphetho

Beha mola oa motheo ka holim'a ntlha e phahameng ka ho fetisisa ea mothinya oa fluorescent oa taolo e mpe.

4.Ho laola boleng

4.1 Taolo e fosahetseng: Ha ho boleng ba Ct bo fumanoeng ho FAM, HEX/VIC channel, kapa Ct＞40;

4.2 Taolo e ntle: Ho FAM, HEX/VIC kanale, Ct≤40;

4.3 Litlhoko tse ka holimo li lokela ho khotsofatsoa tekong e tšoanang, ho seng joalo liphetho tsa liteko li fosahetse 'me teko e lokela ho phetoa.

【Khaola boleng】

Mohlala o nkoa e le o motle ha: Tatellano ea sepheo Ct≤40, The internal control gene Ct≤40.

【Liphetho tlhaloso】

Hang ha taolo ea boleng e fetisitsoe, basebelisi ba lokela ho hlahloba hore na ho na le curve ea amplification bakeng sa sampole ka 'ngoe ho kanale ea HEX/VIC, haeba e teng le ka Ct≤40, e bonts'itse hore mofuta oa taolo ea kahare o matlafalitsoe ka katleho mme tlhahlobo ena e nepahetse.Basebelisi ba ka tsoela pele ho etsa tlhahlobo e latelang:

3. Bakeng sa disampole tse nang le matlafatso ea gene ea taolo ea kahare e hlotsoe (HEX/VIC

channel, Ct＞40, kapa no amplification curve), low Viral load kapa ho ba teng ha PCR inhibitor e ka ba lebaka la ho hlōleha, tlhahlobo e lokela ho phetoa ho tsoa pokellong ea mohlala;

4.Bakeng sa disampole tse ntle le kokoana-hloko e hlahisitsoeng, liphello tsa taolo ea ka hare ha li ame;

Bakeng sa disampole tse fumanweng di se na HIV, taolo ya kahare e lokela ho lekolwa e le ntle, ho seng jwalo sephetho se fosahetse, mme tlhahlobo e lokela ho phetwa, ho qala ka mohato wa pokello ya mohlala.