Kumenya virusi Nucleic Acide

Imiterere ya genomic ya virusi nyinshi yarazwi.Nucleic acide igizwe nibice bigufi bya ADN yagenewe kuvanga hamwe na ADN yuzuzanya cyangwa ibice bya RNA.Urunigi rwa polymerase (PCR) nubuhanga bukora neza mugutahura virusi.Uburyo bwo kwisuzumisha buhanitse bwateguwe vuba aha.

A. Ubuhanga bwo kuvanga aside nucleique

Nucleic aside ivanga cyane cyane harimo Amajyepfo (Amajyepfo) na Amajyaruguru Amajyaruguru (Amajyaruguru), ni tekinike nshya yihuta mu rwego rwo gusuzuma virusi.Impamvu yo kuvanga imvange ni ugukoresha ibice bigufi bya ADN (bita “probe”) bigamije guhuza hamwe na ADN yuzuzanya cyangwa ibice bya RNA.Mu gushyushya cyangwa kuvura alkaline, intego ebyiri ADN cyangwa RNA bigabanijwemo umurongo umwe hanyuma bigahagarikwa ku nkunga ikomeye.Nyuma yibyo, iperereza ryongeweho kandi rivangwa hamwe na ADN cyangwa RNA.Nkuko iperereza ryanditseho isotope cyangwa nuclide idafite radiyo, intego ya ADN cyangwa RNA irashobora kumenyekana binyuze muri autoradiography cyangwa na biotin-avidin.Kubera ko genoside nyinshi za virusi zagiye zikoronizwa kandi zikurikiranwa, zirashobora kumenyekana ukoresheje virusi yihariye nkuko byakurikiranwe.Kugeza ubu, uburyo bwo kuvanga burimo: akadomo, mu guhuza imiterere mu ngirabuzimafatizo, guhuza ADN (ADN) (Amajyepfo) hamwe na RNA (RNA) (Amajyaruguru).

Ikoranabuhanga rya B.PCR

Mu myaka yashize, hakozwe uburyo bwa tekinike ya vitro nucleic aside amplificateur yakozwe hifashishijwe PCR, kugirango isuzume virusi zitumva cyangwa zidahingwa.PCR nuburyo bushobora guhuza urutonde rwa ADN ikurikirana muri vitro polymerase reaction.Inzira ya PCR ikubiyemo uruziga rwubushyuhe rwintambwe eshatu: gutandukana, kwizirika, no kwaguka Ku bushyuhe bwo hejuru (93 ℃ ~ 95 ℃), ADN ifite imirongo ibiri itandukanijwemo imirongo ibiri ya ADN imwe;hanyuma ku bushyuhe buke (37 ℃ ~ 60 ℃), intangiriro ebyiri za nucleotide primers anneal kubice bya ADN byuzuzanya;mugihe ku bushyuhe bukwiye bwa enzyme ya Taq (72 ℃), synthesis yiminyururu mishya ya ADN itangirira kuri primer 3'end ukoresheje ADN yuzuzanya nkicyitegererezo na nucleotide imwe nkibikoresho.Nyuma ya buri cyiciro, urunigi rumwe rwa ADN rushobora kwongerwa muminyururu ibiri.Gusubiramo iki gikorwa, buri munyururu wa ADN ukomatanyirijwe mukuzingo umwe urashobora gukoreshwa nkicyitegererezo mugihe gikurikiraho, kandi umubare wiminyururu ya ADN wikubye kabiri muri buri cyiciro, bivuze ko umusaruro wa PCR wongerewe umuvuduko wa 2n.Nyuma yizunguruka 25 kugeza 30, umusaruro wa PCR ugaragazwa hakoreshejwe electrophore, kandi ibicuruzwa bya ADN byihariye bishobora kugaragara munsi yumucyo UV (254nm).Kubwinyungu zihariye, ibyiyumvo, kandi byoroshye, PCR yemejwe mugupima indwara zanduye virusi nka HCV, VIH, CMV, na HPV.Nkuko PCR yunvikana cyane ， irashobora kumenya virusi ya ADN kurwego rwa fg, igikorwa kigomba gukorwa neza cyane kugirango wirinde ibyiza.Byongeye kandi, ibisubizo byiza mugupima aside nucleic ntabwo bivuze ko muri virusi harimo virusi yanduye.

Hamwe nogukoresha kwinshi tekinike ya PCR, tekiniki nuburyo bushya byatejwe imbere hashingiwe kubuhanga bwa PCR kubwintego zitandukanye.Kurugero, igihe nyacyo PCR ishobora kumenya imitwaro ya virusi;mumwanya PCR ikoreshwa mukumenya kwandura virusi mubice cyangwa selile;PCR yubatswe irashobora kongera umwihariko wa PCR.Muri byo, igihe nyacyo PCR yatejwe imbere byihuse.Ubuhanga bwinshi bushya, nka TaqMan hydrolysis probe, Hybridisation probe, hamwe na molekuline ya beacon probe, byahujwe muburyo nyabwo bwa PCR igereranya, ikoreshwa cyane mubushakashatsi bwamavuriro.Usibye kumenya umutwaro wa virusi mumazi yumubiri wabarwayi, ubu buryo burashobora no gukoreshwa mugutahura imiti yihanganira ibiyobyabwenge.Kubwibyo, igihe nyacyo PCR ikoreshwa cyane mugusuzuma ingaruka zikiza no kugenzura kwihanganira ibiyobyabwenge.

C. Kumenyekanisha-hejuru ya virusi nucleic acide

Kugira ngo hakemurwe ibisubizo byihuse by’indwara nshya zandura, hashyizweho uburyo butandukanye bwo gutahura ibintu byinshi, nka ADN chip (ADN).Kuri chip ya ADN, iperereza ryihariye rirahuzwa kandi rigafatanwa na chipo ntoya ya silicon mubucucike bwinshi cyane kugirango habeho micarray ya ADN (ADN) ishobora kuvangwa nicyitegererezo.Ikimenyetso cyo kuvanga kirashobora gushushanywa na microscope ya conocal cyangwa laser scaneri hanyuma bigatunganywa na mudasobwa kandi amakuru manini yashyizweho yerekeye genes zitandukanye arashobora kuboneka.Hariho ubwoko bubiri bwa chip ya ADN."Synthesis chip" ni nkibi bikurikira: oligonucleotide yihariye ikomatanyirizwa kuri chip.Ikindi ni chip ya ADN.Imirasire ya clone cyangwa ibicuruzwa bya PCR byacapishijwe neza kurupapuro.Ibyiza bya tekinoroji ya chip ya ADN ni icyarimwe kumenya icyarimwe umubare munini wa ADN ikurikirana.Verisiyo iheruka ya chip yamenyekanye irashobora kumenya virusi zirenga 1700 icyarimwe.Ikoranabuhanga rya chip chip ya ADN ryakemuye ibibazo byuburyo bwa Hybridiside ya nucleique gakondo kandi bifite uburyo bwagutse cyane mugupima virusi no kwiga epidemiologiya.