Monkeypox Virus (MPV) Nucleic Acid Detection Kit

Iyo kit ine ma reagents ekugadzirisa makumi mana nemasere bvunzo kana kudzora kwemhando, kusanganisira zvinotevera zvikamu:

Reagent A

ReagentB

Ongorora: Izvo zvikamu zvenhamba dzebatch dzakasiyana hazvigone kushandiswa zvakasiyana

【Storage Mamiriro uye Sherufu Hupenyu】

1.Reagent A / B inogona kuchengetwa pa 2-30 ° C, uye hupenyu hwesherufu ndeye 10 mwedzi.

2.Ndapota vhura test tube cover chete kana wagadzirira bvunzo.

3.Usashandisa test tubes kupfuura zuva rekupera.

4.Usashandisa chubhu yekuona inodonha.

【Applicable Instrument】

Inokodzera Inokodzera LC480 PCR yekuongorora system, Gentier 48E Automatic PCR yekuongorora system, ABI7500 PCR yekuongorora system.

【Mienzaniso Zvinodiwa】

1.Inoshanda mhando dzemhando: huro swabs samples.

2.Sampling mhinduro:Mushure mekuona, zvinokurudzirwa kushandisa yakajairika saline kana Virus kuchengetedza chubhu inogadzirwa neHangzhou Testsea biology yekuunganidza sampuli.

pahuro swab:pukuta bilateral pharyngeal tonsils uye posterior pharyngeal madziro nekuraswa sterile sampling swab, nyudza swab muchubhu ine 3mL sampling solution, kurasa muswe, uye sunga chubhu chivharo.

3.Sample kuchengetedza uye kutumira:Mienzaniso inofanirwa kuongororwa inofanira kuongororwa nekukurumidza sezvinobvira.Kupisa kwekutakura kunofanira kuchengetwa pa 2 ~ 8 ℃. Masampuli anogona kuongororwa mukati meawa makumi maviri nemana anogona kuchengetwa pa 2 ℃ ~ 8 ℃ uye kana samples isingagoni kuongororwa mukati maawa makumi maviri nemana, inofanira kuchengetwa zvishoma kudarika kana kuenzana. kusvika -70 ℃ (kana pasina chekuchengetedza -70 ℃, inogona kuchengetwa pa -20 ℃ kwenguva pfupi), dzivisa kudzokororwa

kutonhora nekunyunguduka.

4.Kutora sampuli yakakodzera, kuchengetedza, uye kutakura zvakakosha pakushanda kwechigadzirwa ichi.

【Testing Method】

1.Sample processing uye sampuli yekuwedzera

1.1 Sample processing

Mushure mekusanganisa mhinduro yesampling iri pamusoro nemasample, tora 30μL yemuenzaniso muDNA inoburitsa reagent chubhu wosanganisa zvakaenzana.

1.2 Loading

Tora 20μL ye reconstitution reagent woiwedzera kuMPV yekuona reagent, wedzera 5μL yemuenzaniso wakagadziridzwa uri pamusoro (Kudzora kwakanaka uye kutonga kwakashata kuchagadziriswa nenzira yakafanana nemasampuli), vhara chivharo chechubhu, centrifuge pa 2000rpm kwegumi. seconds.

2. PCR kuwedzera

2.1 Takura iyo yakagadzirirwa PCR ndiro / machubhu kune fluorescence PCR chiridzwa, Negative control uye kutonga kwakanaka kuchaiswa kune yega bvunzo.

2.2 Fluorescent chiteshi kugadzirisa:

1)Sarudza chiteshi cheFAM chekuona MPV;

2)Sarudza HEX/VIC chiteshi chemukati chekudzora gene kuona;

3.Kuongorora kwemigumisiro

Seta mutsara wepasi pamusoro penzvimbo yepamusoro yeiyo negative control's fluorescent curve.

4.Quality control

4.1 Negative control: Hapana Ct kukosha yaonekwa muFAM, HEX/VIC chiteshi, kana Ct＞40;

4.2 Positive control: MuFAM, HEX/VIC chiteshi, Ct≤40;

4.3 Zvinodiwa zviri pamusoro zvinofanirwa kugutsikana mukuyedza kumwe chete, zvikasadaro mhedzisiro yebvunzo haina basa uye kuyedza kunoda kudzokororwa.

【Bvisa kukosha】

Sample inoonekwa seyakanaka kana: Target sequence Ct≤40, The internal control gene Ct≤40.

【Dudziro yemigumisiro】

Kana kutonga kwemhando kwapfuura, vashandisi vanofanirwa kutarisa kana paine amplification curve yesample yega yega muHEX/VIC chiteshi, kana iripo uye neCt≤40, yakaratidza kuti yemukati yekudzora gene yakakwidziridzwa zvinobudirira uye iyi bvunzo inoshanda.Vashandisi vanogona kuenderera kune yekutevera ongororo:

3.Kumasamples ane kukwidziridzwa kwejene rekutonga kwemukati zvakundikana (HEX/VIC

chiteshi, Ct＞40, kana pasina amplification curve), yakaderera Viral load kana kuvepo kwePCR inhibitor kunogona kunge kuri chikonzero chekutadza, bvunzo dzinofanira kudzokororwa kubva mukuunganidzwa kwemuenzaniso;

4.Kune sampuli dzakanaka uye hutachiwana hwakagadzirwa, migumisiro yekutonga kwemukati haibatsiri;

Kune masampuli akaongororwa kuti haana, kutonga kwemukati kunoda kuongororwa kuti ane positive zvikasadaro mhedzisiro yacho haina basa uye bvunzo inoda kudzokororwa, kutanga kubva padanho rekuunganidza.