ʻO Coronavirus hou(SARS-Cov-2) pahu ʻimi ʻakika nucleic

New Coronavirus(SARS-Cov-2) Nucleic Acid Detection Kit

(Fluokascent RT-PCR Probe Method) Product Manual

【Product name 】ʻO ka Coronavirus hou(SARS-Cov-2) ka pahu ʻike ʻakika nucleic (Fluorescent RT-PCR Probe Method)

【Packaging specifications 】25 Ho'āʻo/Kiki

【Intended usmakahiki】

Hoʻohana ʻia kēia kit no ka ʻike qualitative o ka waikawa nucleic mai ka coronavirus hou i loko o nā swabs nasopharyngeal, oropharyngeal (puʻi) swabs, nā ʻūhā ihu mua, nā swabs waena waena, nā holoi ihu a me nā aspirates ihu mai nā poʻe i manaʻo ʻia he COVID19 e kā lākou mea mālama olakino. Hiki ke hoʻohana ʻia ka ʻike ʻana i nā genes ORF1ab a me N o ka coronavirus hou no ka maʻi kōkua a me ka nānā ʻana i ka maʻi epidemiological o ka maʻi coronavirus hou.

【Principles of the procedure 】

Hoʻolālā ʻia kēia kime no nā ʻimi kikoʻī TaqMan a me nā kumu mua i hoʻolālā ʻia no ka novel coronavirus (SARS-Cov-2) ORF1ab a me N gene sequences. Aia i loko o ka solution reaction PCR he 3 sets o nā primers kiko'ī a me nā ʻimi fluorescent no ka ʻike pono ʻana i nā pahuhopu, a ua hoʻohana ʻia kekahi pūʻulu o nā primers kikoʻī a me nā ʻimi fluorescent ma ke ʻano he mana maʻamau o loko o ka pahu no ka ʻike ʻana i nā genes mālama hale endogenous.

ʻO ke kumu hoʻāʻo ʻo ia ka hoʻokaʻawale ʻana a hoʻohaʻahaʻa ʻia ka ʻimi fluorescent kikoʻī e ka hana exonuclease o ka enzyme Taq i ka hopena PCR, i hoʻokaʻawale ʻia ka pūʻulu fluorescent reporter a me ka hui fluorescent quenched, i hiki i ka ʻōnaehana nānā fluorescence ke loaʻa i kahi fluorescent. hōʻailona, ​​a laila Ma o ka hoʻonui ʻia ʻana o ka PCR amplification, hiki i ka hōʻailona fluorescence o ka probe i kahi waiwai paepae-Ct waiwai (Cycle threshold). Inā ʻaʻohe amplicon i hoʻopaʻa ʻia, kokoke ka pūʻulu hoʻolaha o ka probe i ka hui quenching. I kēia manawa, hiki ke hoʻololi i ka ikehu fluorescence resonance, a ua kinai ʻia ka fluorescence o ka hui reporter e ka hui quenching, i hiki ʻole ke ʻike ʻia ka hōʻailona fluorescent e ka mea hana PCR fluorescent.

No ka nānā ʻana i ka hoʻohana ʻana i nā reagents i ka wā o ka hoʻāʻo ʻana, ua lako ka pahu me nā mana maikaʻi a maikaʻi ʻole: aia ka mana maikaʻi i ka plasmid recombinant pūnaewele i manaʻo ʻia, a ʻo ka mana maikaʻi ʻole ʻo ka wai Distilled, i hoʻohana ʻia e nānā i ka pollution kaiapuni. Manaʻo ʻia e hoʻonohonoho i kahi mana maikaʻi a me kahi mana maikaʻi ʻole i ka manawa like i ka wā e hoʻāʻo ai.

【Main components 】

Nānā：(1) ʻAʻole hiki ke hoʻohui a hoʻololi ʻia nā ʻāpana o nā pahu pahu like ʻole.

(2) E hoʻomākaukau i kāu reagent ponoʻī: Nucleic acid extraction kit.

【Storage conditions a expiration date 】

For BST-SARS-25:ʻO ka lawe a mālama ʻana ma -20±5 ℃ no ka manawa lōʻihi.

For BST-SARS-DR-25:ʻO ka lawe ʻana i ka lumi wela. E kūʻai ma -20±5 ℃ no ka manawa lōʻihi.

E hōʻalo i nā pōʻai hoʻoheheʻe-hau. Hoʻonohonoho ʻia ka manawa kūpono no 12 mau mahina.

E ʻike i ka lepili no ka lā hana a hoʻohana.

Ma hope o ka wehe mua ʻana, hiki ke mālama ʻia ka reagent ma -20 ± 5 ° C no ka mea ʻaʻole i ʻoi aku ma mua o 1 mahina a i ʻole ka pau ʻana o ka manawa reagent, ʻo kēlā me kēia lā e hiki mua mai, e pale i nā pōʻai manuahi-thaw hou, a me ka helu o ka reagent freeze. -ʻaʻole pono eʻoi aku ka nui o nā manawa hoʻoheheʻe ma mua o 6 manawa.

【Applicable instrument】ABI 7500, SLAN-96P, Roche-LightCycler-480.

【Sample requirements 】

1.Applicable laʻanaʻano: Extracted nucleic acid solution.

2.Sample mālama a me ka lawe ʻana: E kūʻai ma-20±5 ℃ no 6 mau mahina. E hoʻokuʻu a hoʻoheheʻe i nā laʻana ma mua o 6 mau manawa.

【Testing method】

1.Nucleic acid extraction

E koho i kahi pahu lawe ʻakika nucleic kūpono no ka unuhi ʻana i ka waika nucleic viral, a e hahai i nā ʻōlelo aʻoaʻo e pili ana. Manaʻo ʻia e hoʻohana i kahi pahu hoʻomaʻemaʻe Nucleic acid i hana ʻia e Yixin Bio-Tech (Guangzhou) Co., Ltd. a i ʻole ka pahu hoʻomaʻemaʻe nucleic acid.

2.  Reaction reagent preparation

2.1 For BST-SARS-25:

(1) Wehe i ka SARS-Cov-2 Mix a me Enzyme Mix , hehee loa i ka lumi wela e hui pono me ka Vortex a laila centrifuge pōkole.

(2) Hoʻohui ʻia ʻo 16.5uL Enzyme Mix i ka 358.5uL SARS-Cov-2 Mix a hui maikaʻi ʻia e loaʻa ai ka hopena hoʻohuihui.

(3) E hoʻomākaukau i kahi paipu octal PCR maʻemaʻe 0.2 mL a e kaha iā ia me 15uL o ka hoʻonā hui ʻia ma luna o kēlā me kēia punawai.

(4) E hoʻohui i 15 μL o ka hoʻomaʻemaʻe nucleic acid solution, ka mana maikaʻi a me ka mana maikaʻi ʻole, a uhi pono i ka pāpale octal tube.

(5) Hoʻohui maikaʻi ma ka hoʻohuli ʻana i luna i lalo, a e hoʻokaʻawale wikiwiki i ka wai ma lalo o ka paipu.

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