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ʻO ka PCR pololei ka hopena e hoʻohana pono ana i nā ʻiʻo holoholona a mea kanu paha no ka hoʻonui ʻana me ka ʻole o ka unuhi ʻana o ka nucleic acid.Ma nā ʻano he nui, hana ka PCR pololei e like me ka PCR maʻamau

ʻO ka ʻokoʻa nui, ʻo ia ka pale maʻamau i hoʻohana ʻia i ka PCR pololei, hiki ke hoʻopili pololei ʻia ka laʻana i ka hopena PCR me ka ʻole o ka ʻokiʻoki nucleic acid, akā aia nā koi kūpono no ka hoʻomanawanui ʻana o nā enzymes a me ka hoʻohālikelike o ka buffer i komo i ka hopena PCR pololei.

ʻOiai he nui a liʻiliʻi paha ka PCR inhibitors i nā laʻana maʻamau, hiki i ka PCR pololei ke hoʻokō i ka hoʻonui hilinaʻi ma lalo o ka hana a nā enzymes a me nā buffers.Pono ka hoʻololi PCR kuʻuna i ka waika nucleic kiʻekiʻe ma ke ʻano he laʻa, hiki ke pale i ka holomua o ka hoʻololi PCR inā loaʻa nā protein a me nā mea haumia ʻē aʻe.ʻO Direct PCR i kēia manawa kekahi o nā ʻenehana kaulana loa i ke kahua o ka diagnostics molecular.

01 Ua hoʻohana mua ʻia ʻo Direct PCR no nā holoholona a me nā mea kanu

ʻO ka noi mua loa o ka PCR pololei ma ke kula o nā holoholona a me nā mea kanu, e like me ke koko, ka ʻiʻo a me ka lauoho o ka ʻiole, pōpoki, moa, rabbit, hipa, pipi, etc., nā lau kanu a me nā ʻanoʻano, etc., i hoʻohana ʻia e aʻo i ka genotyping, transgenic, plasmid detection, Gene knockout analysis, DNA source identification, species identification, SNP analysis and other field.

Loaʻa i kēia mau māla kekahi mau hiʻohiʻona maʻamau, ʻo ia hoʻi, ʻoi aku ka kiʻekiʻe o ka maʻiʻo o ka gene a me ka hoʻoneʻe nucleic acid he pilikia, no laila ʻaʻole hiki i ka PCR pololei ke mālama i ka manawa a loaʻa ka hopena liʻiliʻi i nā hopena, akā mālama pū kekahi i ke kumukūʻai.

ʻO ka PCR pololei i hoʻohana ʻia no ka ʻike pathogen he mea ia o nā makahiki i hala iho nei, ua hana nui nā mea hana PCR reagent i kēia ala i ka wā e hana ai i ka hana hou.ʻOi loa i kēia maʻi maʻi COVID-19, ua ʻike ʻia nā huahana ʻike he nui ma ka mākeke, e like me ka SARS-CoV-2 Nucleic Acid Detection Kit (Multiplex PCR Fluorescent Probe Method) i noiʻi ʻia a hoʻomohala ʻia e Foregene, e hoʻohana ana i ka ʻenehana RT PCR manawa maoli (rRT-PCR) no ka ʻike qualitative o ka swasharyngeal-2 o ke kanaka nucleic acid i loko o ke kanaka swa-CoVharyngeal-2 nucleic acid.

ʻO Foregene kekahi o nā hui e hoʻohana ana i ka ʻenehana Direct PCR, no ka ʻike ʻana i ka ORF1ab maʻamau, N, E, a meʻokoʻa laina nucleic acids i loko o ke kanaka nasopharyngeal a oropharyngeal swab samples like SARS-CoV-2 B.1.1.7 lineage (UK), B.1.351 lineage (ZA), B.1.617 lineage (IND) and P.1 lineage (BR).

02  Pono nā reagents no ka PCR pololei

Laʻana Lysate

Hiki ke hoʻonohonoho ʻia ka laʻana lysate e ʻoe a kūʻai ʻia paha.ʻO ka ʻokoʻa o ka haku mele ʻana o nā ʻano like ʻole o ka lysate e hoʻokaʻawale i ka hiki lysing, a laila e ʻokoʻa iki ka manawa lysing.No ka laʻana, no ka hoʻomākaukau ʻana i nā laʻana o nā ʻiʻo holoholona, ​​ʻo 30 mau minuke a i ʻole lysis i ka pō e ʻōlelo pinepine ʻia, a ʻo ka hopena lysis no nā maʻi mai ka 3-10 mau minuke.

PCR haku hui

Manaʻo ʻia e hoʻohana i ka polymerase DNA hoʻomaka wela e hoʻomaikaʻi i ka amplification kikoʻī a hoʻonui i ka mana amplification.ʻO ke kumu o ka PCR pololei he polymerase hoʻomanawanui loa.

Hoʻopau a keʻakeʻa paha i nā ʻāpana o ka laʻana e pili ana i ka hoʻonui ʻana i ka DNA

Ma hope o ka hana ʻia ʻana o ka hāpana me ka lysate, e hoʻokuʻu ʻia nā protein, lipids a me nā ʻōpala cell ʻē aʻe, e kāohi kēia mau mea i ka hopena PCR.No laila, pono ka PCR pololei i ka hoʻohui ʻana o ka wehe ʻana a i ʻole nā ​​mea paʻa e hōʻemi i ka mana o kēia mau kumu.

03  He hōʻiliʻili o ʻelima mau ʻike o ka PCR pololei

ʻO ka mua, ʻo ka ʻenehana PCR pololei he ʻenehana PCR pololei no nā ʻano laʻana olaola like ʻole.Ma lalo o kēia kūlana ʻenehana, ʻaʻohe pono e hoʻokaʻawale a unuhi i ka waikawa nucleic, e hoʻohana pololei i ka ʻiʻo kikoʻī ma ke ʻano he mea, a hoʻohui i nā kumu kumu kumu e hana ai i ka hopena PCR.

ʻO ka lua, ʻo ka ʻenehana PCR Direct ʻaʻole ia he ʻenehana hoʻonui hoʻohālikelike DNA kuʻuna, akā aia pū kekahi RNA template reverse transcription PCR.

ʻO ke kolu, ʻaʻole hana pololei ka ʻenehana PCR pololei i nā hopena PCR qualitative maʻamau ma nā ʻāpana kiko, akā e komo pū kekahi i nā hopena qPCR Real-Time, kahi e koi ai i ka ʻōnaehana hoʻonā e loaʻa i ka ikaika anti-background fluorescence interference a me endogenous fluorescence quenchers i ka antagonistic hiki.

ʻO ka hā, ʻo nā laʻana i manaʻo ʻia e ka ʻenehana Direct PCR pono wale nō e hoʻokuʻu i nā mamana nucleic acid, a ʻaʻole e wehe i nā proteins, polysaccharides, salt ions, etc.Pono ia i ka nucleic acid polymerase a me ka PCR Mix i loko o ka ʻōnaehana hopena e loaʻa i ke kūpaʻa maikaʻi a me ka hoʻololi ʻana e hōʻoia i ka hana enzyme a me ka pololei o ka hana hou ʻana ma lalo o nā kūlana paʻakikī.

ʻO ka lima, ʻo ka hāpana kikoo i manaʻo ʻia e ka ʻenehana Direct PCR me ka ʻole o ka mālama ʻana i ka waikawa nucleic a he liʻiliʻi loa ka nui o ka template, kahi e koi ai i ka ʻōnaehana hopena e loaʻa i ka ʻike kiʻekiʻe a me ka hoʻonui ʻana.


Ka manawa hoʻouna: Iune-28-2021