ʻO ka Monkeypox Virus Kiko PCR Manawa maoli

1. LaʻanaʻOhi:Pono e hōʻiliʻili ʻia nā mea hoʻohālike i loko o nā paipu sterile e like me

me nā kikoʻī loea maʻamau.

2. Hoʻomākaukau Reagent (Reagent Preparation Area)

Wehe i nā ʻāpana o ka pahu, hoʻohālikelike iā lākou i ka wela lumi no ka hoʻohana ʻana i ka standby.

3. Ka Hana Hana ʻana (Wāhi Hana Hana ʻia)

3.1 ʻO ka unuhi ʻana i ka ʻakika nucleic

Manaʻo ʻia e lawe i 200μL wai samples, Positive Control and Negative Control no ka nucleic acid extraction, e like me nā koi kūpono a me nā kaʻina hana o nā pahu hoʻoiho DNA viral.

3.2 Hoʻoheheʻe ʻia ka pauka Lyophilized a me ka hoʻohui kumu

E hoʻomākaukau i ka Monkeypox Virus Lyophilized premix e like me ka helu o nā laʻana.Pono kekahi laʻana i hoʻokahi paipu PCR i loko o ka pauka premix Lyophilized.Pono e mālama ʻia ka mana maikaʻi ʻole a me ka mana maikaʻi e like me nā laʻana ʻelua.

(1) Hoʻohui i ka 15μL Dissolving Solution i loko o kēlā me kēia paipu PCR i loaʻa i ka Lyophilized premix, a laila hoʻohui i 5μL i unuhi ʻia nā laʻana / Mana Manaʻo / Manaʻo maikaʻi i kēlā me kēia paipu PCR.

(2) E uhi paʻa i nā paipu PCR, e paʻi lima i nā paipu PCR a hiki i ka pau ʻana o ka pauka lyophilized a hui pū ʻia, e hōʻiliʻili i ka wai i lalo o nā paipu PCR e ka centrifugation haʻahaʻa haʻahaʻa.

(3) Inā hoʻohana maʻamau i ka mea hana PCR manawa maoli no ka ʻike ʻana, a laila e hoʻololi pololei i nā paipu PCR i ka wahi hoʻonui;inā hoʻohana i ka BTK-8 no ka ʻike ʻana, a laila pono e hana i kēia mau hana: e hoʻololi i ka wai 10 μL mai ka paipu PCR i ka puʻupuʻu maikaʻi o BTK-8.Hoʻokahi paipu PCR e pili ana i hoʻokahi punawai ma ka chip.I ka hana ʻana i ka pipetting, e hōʻoia i ka 90 degere ke kū pololei ka pipette.Pono e kau ʻia nā ʻōlelo aerosol barrier pipette i waenakonu o ka pūnāwai me ka ikaika haʻahaʻa a hoʻōki i ka hoʻokuke ʻana i ka pipette ke hiki i ka mea hana mua (e pale i nā ʻōhū).Ma hope o ka hoʻopiha ʻia ʻana o nā pūnāwai, e lawe i kahi membrane chip reaction no ka uhi ʻana i nā pūnāwai a pau a laila hoʻoneʻe ʻia ka chip i ka wahi ʻike hoʻonui.

4. Hoʻonui PCR (Wahele ʻIke)

4.1 E hoʻokomo i nā paipu PCR/rection chip i loko o ka pahu hopena a hoʻonohonoho pono i nā inoa o kēlā me kēia pane ma ke ʻano kūpono.

4.2 Nā hoʻonohonoho o ka ʻike fluorescence: (1) Monkeypox virus (FAM);(2) Mana Kūloko (CY5).

4.3 E holo i kēia kaʻina kaʻa kaʻa

Kūkākūkā o ABI7500, Bio-Rad CFX96, SLAN-96S, QuantStudio:

 Kaʻina o BTK-8:

5. Ka nānā 'ana i nā hualoa'a (e nānā i ka Manual User Manual)

Ma hope o ka hopena, e mālama ʻia nā hopena.E kaomi i ka "Analyze" no ke kālailai ʻana, a na ka mea hana e wehewehe ʻakomi i nā waiwai Ct o kēlā me kēia laʻana ma ke kolamu hopena.ʻO nā hopena hoʻomalu maikaʻi ʻole a me nā hopena hoʻomalu maikaʻi e kūlike me kēia "6. Mana Mana maikaʻi ".

6. Ka Mana Mana

6.1 Manaʻo maikaʻi ʻole: ʻAʻohe Ct a i ʻole Ct> 40 ma ke kahawai FAM, Ct≤40 ma ke kahawai CY5 me ka pihi hoʻonui maʻamau.

6.2 Manaʻo maikaʻi: Ct≤35 i ke kahawai FAM me ka pihi hoʻonui maʻamau, Ct≤40 i ke kahawai CY5 me ka pihi hoʻonui maʻamau.

6.3 Pono ka hopena inā e hoʻokō ʻia nā pae hoʻohālike a pau.A i ʻole, ʻaʻole kūpono ka hopena.

Huna Hula

Hiki i kēia mau hopena:

HOOLAHA: Inā loaʻa ka hopena kūpono ʻole, pono e hōʻiliʻili a hoʻāʻo hou ka hāpana.