Ua ʻike ʻia nā kaʻina genomic o ka hapa nui o nā maʻi.Nucleic acid probes he mau ʻāpana pōkole o ka DNA i hoʻolālā ʻia e hoʻohui me nā ʻāpana viral viral a i ʻole RNA.ʻO ka polymerase chain reaction (PCR) kahi hana ʻoi aku ka maikaʻi no ka ʻike viral.Ua kūkulu ʻia nā ʻano diagnostic high throughput i kēia manawa.
A. Nucleic acid hybridization technique
ʻO ka hoʻohuihui ʻana o ka waikawa Nucleic, ka hapa nui me ka Southern blotting (Southern) a me ka Northern blotting (Northern), kahi hana wikiwiki e hoʻomohala hou i ka kahua diagnostic virus.ʻO ke kumu o ka hoʻohālikelike ʻana ʻo ka hoʻohana ʻana i nā ʻāpana pōkole o ka DNA (i kapa ʻia ʻo "probe") i hoʻolālā ʻia e hoʻohui me nā ʻāpana viral viral a i ʻole RNA.Ma ka hoʻomehana ʻana a i ʻole ka hoʻomaʻamaʻa alkaline, ua hoʻokaʻawale ʻia ʻo DNA a i ʻole RNA pae ʻelua i nā kaula hoʻokahi a laila hoʻopaʻa ʻia ma ke kākoʻo paʻa.Ma hope o kēlā, hoʻohui ʻia ka probe a hoʻohui ʻia me ka DNA a i ʻole RNA.Ke hoʻopaʻa inoa ʻia ka probe me ka isotope a i ʻole ka nuclide non-radioactive, hiki ke ʻike ʻia ka DNA a i ʻole RNA i ʻike ʻia ma o autoradiography a i ʻole e ka biotin-avidin system.No ka mea, ua hoʻopili ʻia ka hapa nui o nā genome viral, hiki ke ʻike ʻia me ka hoʻohana ʻana i nā kaʻina virus-specific ma ke ʻano he probes i ka specimen.I kēia manawa, loaʻa nā ʻano hana hoʻohuihui: dot blot , in situ hybridization in cell , DNA blotting(DNA) (Southern blot) a me RNA blotting(RNA) (Northern blot).
ʻenehana B.PCR
I nā makahiki i hala iho nei, ua hoʻomohala ʻia kahi ʻano o ka in vitro nucleic acid amplification techniques e pili ana i ka PCR, e hoʻāʻo ai i nā maʻi maʻi insensitive a i ʻole kanu ʻia.ʻO ka PCR kahi ala e hiki ai ke hoʻohui i ke kaʻina DNA kikoʻī ma o ka hopena in vitro polymerase.ʻO ke kaʻina hana o ka PCR e loaʻa i kahi pōʻai wela o ʻekolu mau ʻanuʻu: denaturation , annealing , a me ka hoʻonui I ka wela kiʻekiʻe (93 ℃ ~ 95 ℃), ua hoʻokaʻawale ʻia ka DNA stranded pālua i ʻelua mau kaula DNA hoʻokahi;a laila ma ka wela haʻahaʻa (37 ℃ ~ 60 ℃), ʻelua mau kumu nucleotide synthesized e hoʻopili i nā ʻāpana DNA hoʻohui;ʻoiai ma ka mahana kūpono no Taq enzyme (72 ℃), hoʻomaka ka synthesis o nā kaulahao DNA hou mai ka 3' hope me ka hoʻohana ʻana i ka DNA hoʻohui ma ke ʻano he mau mea hoʻohālike a me nā nucleotides hoʻokahi i mea.No laila ma hope o kēlā me kēia pōʻai, hiki ke hoʻonui ʻia kahi kaulahao DNA i ʻelua kaulahao.Ma ka hana hou ʻana i kēia kaʻina hana, hiki ke hoʻohana ʻia kēlā me kēia kaulahao DNA i hoʻohui ʻia i hoʻokahi pōʻaiapuni ma ke ʻano he hoʻohālikelike i ka pōʻaiapuni aʻe, a ua pālua ʻia ka helu o nā kaulahao DNA i kēlā me kēia pōʻai, ʻo ia hoʻi, ua hoʻonui ʻia ka hana ʻana o PCR i ka wikiwiki 2n log.Ma hope o 25 a 30 mau pōʻai, ʻike ʻia ka hana ʻana o PCR ma o ka electrophoresis, a hiki ke nānā ʻia nā huahana DNA kikoʻī ma lalo o ke kukui UV (254nm).No ka pono o ka kiko'ī, ka naʻau, a me ka maʻalahi, ua hoʻohana ʻia ka PCR i ka hōʻoia ʻana o nā maʻi maʻi he nui e like me HCV, HIV, CMV, a me HPV.Ma muli o ka maʻalahi o ka PCR, hiki iā ia ke ʻike i ka DNA virus ma ka pae fg, pono e mālama pono ʻia ka hana e pale aku i ka maikaʻi ʻole.Eia kekahi, ʻaʻole manaʻo ka hopena maikaʻi i ka hoʻāʻo ʻana i ka waika nucleic i loko o ka hāpana.
Me ka hoʻohana ākea o ka ʻenehana PCR, kūkulu ʻia nā ʻenehana hou a me nā ʻano hana e pili ana i ka ʻenehana PCR no nā kumu hoʻāʻo like ʻole.No ka laʻana, hiki i ka PCR quantitative manawa maoli ke ʻike i ka ukana viral;hoʻohana ʻia ka PCR in situ no ka ʻike ʻana i ka maʻi maʻi i loko o ka ʻiʻo a i ʻole nā pūnaewele;Hiki i ka PCR nested ke hoʻonui i ka kikoʻī o PCR.Ma waena o lākou, ʻoi aku ka wikiwiki o ka PCR quantitative manawa maoli.Nui nā ʻano hana hou, e like me TaqMan hydrolysis probe, hybridization probe, a me molecular beacon probe, ua hoʻohui ʻia i loko o ka ʻenehana PCR quantitative maoli, i hoʻohana nui ʻia i ka noiʻi lapaʻau.Ma waho aʻe o ka ʻike pololei ʻana i ka ukana viral i loko o ka wai kino o nā mea maʻi, hiki ke hoʻohana ʻia kēia ala e ʻike ai i ka mutant hoʻomanawanui i ka lāʻau.No laila, hoʻohana nui ʻia ka PCR quantitative maoli i ka loiloi hopena curative a me ka nānā ʻana i ka lāʻau.
C. Kiʻekiʻe-throughput ike o viral nucleic acids
No ka hoʻokō ʻana i nā pono no ka hoʻomaʻamaʻa wikiwiki ʻana i nā maʻi infectious hou, ua hoʻokumu ʻia nā ʻano hana ʻike kiʻekiʻe, e like me DNA chips (DNA).No nā ʻāpana DNA, hoʻohui ʻia nā ʻimi kikoʻī a hoʻopaʻa ʻia i nā ʻāpana silika liʻiliʻi ma ke ʻano kiʻekiʻe loa e hana i ka DNA probe microarray (DNA) hiki ke hoʻohui ʻia me ka laʻana.Hiki ke kiʻi ʻia ka hōʻailona o ka hybridization e ka confocal microscope a i ʻole ka laser scanner a e hoʻoponopono hou ʻia e ke kamepiula a hiki ke loaʻa ka ʻikepili nui e pili ana i nā ʻano genes.ʻElua ʻano o ka chip DNA.ʻO ka "synthesis chip" e like me kēia: ua hoʻopili pololei ʻia nā oligonucleotides kikoʻī ma luna o nā chips.ʻO kekahi mea ʻo DNA pool chip.Paʻi pololei ʻia nā genes cloned a i ʻole nā huahana PCR ma ka slide.ʻO ka pōmaikaʻi o ka ʻenehana chip DNA ka ʻike like ʻana o ka nui o nā kaʻina DNA.ʻO ka mana hou loa o ka puʻupuʻu pathogen detection hiki ke ʻike ma luna o 1700 mau maʻi kanaka i ka manawa hoʻokahi.Ua hoʻoponopono ʻia ka ʻenehana chip DNA i nā pilikia o nā ʻano kuʻuna nucleic acid hybridization a loaʻa nā noi ākea loa i ka maʻi maʻi viral a me ka noiʻi epidemiological.
Ka manawa hoʻouna: Dec-23-2020