PCR itaziguye ni reaction ikoresha mu buryo butaziguye inyamanswa cyangwa ibimera kugirango byongerwe imbaraga nta gukuramo aside nucleic.Muburyo bwinshi, kuyobora PCR ikora nka PCR isanzwe

Itandukaniro nyamukuru ni buffer yihariye ikoreshwa muri PCR itaziguye, icyitegererezo gishobora gukorerwa byimazeyo PCR itavuyemo acide nucleic, ariko haribisabwa bijyanye no kwihanganira imisemburo no guhuza kwa buffer bigira uruhare muburyo butaziguye bwa PCR.

Nubwo hariho byinshi cyangwa bike bya inhibitori ya PCR mubitegererezo bisanzwe, PCR irashobora kugera kuri amplification yizewe ikorwa na enzymes na buffers.Imyitwarire ya PCR isanzwe isaba aside nucleic yo mu rwego rwo hejuru nkicyitegererezo, ishobora kubuza iterambere ryiza rya PCR niba inyandikorugero irimo proteyine nibindi byanduye.Direct PCR kuri ubu ni bumwe mu buhanga buzwi cyane mu bijyanye no gusuzuma molekile.

01 PCR itaziguye yakoreshwaga mubikoko n'ibimera

Ikoreshwa rya mbere rya PCR ritaziguye ni mubijyanye ninyamaswa n’ibimera, nkamaraso, tissue numusatsi wimbeba, injangwe, inkoko, urukwavu, intama, inka, nibindi, amababi yimbuto nimbuto, nibindi, bikoreshwa mukwiga genotyping, transgenic, plasmid detection, Gene knockout, kumenya inkomoko ya ADN, kumenya amoko, gusesengura SNP nizindi nzego.

Iyi mirima ifite ibintu bimwe bihuriweho, ni ukuvuga, intego ya gene igereranijwe ni ndende kandi gukuramo aside nucleic ni ikibazo, bityo PCR itaziguye ntishobora gutakaza umwanya gusa kandi igira ingaruka nke kubisubizo, ariko kandi ikanabika ikiguzi.

PCR itaziguye ikoreshwa mugutahura indwara ni ikibazo cyimyaka yashize, bamwe mubakora PCR reagent bakoze imbaraga nyinshi muriki cyerekezo mugihe bakora udushya.By'umwihariko muri iki cyorezo cya COVID-19, ibicuruzwa byinshi byo gutahura byagaragaye ku isoko, nka SARS-CoV-2 Nucleic Acide Detection Kit (Multiplex PCR Fluorescent Probe Method) byakorewe ubushakashatsi kandi bikozwe na Foregene, ikoresha ikoranabuhanga rya RT-PCR (rRT-PCR) kugira ngo hamenyekane neza SARS-CoV-2 ya nucleopique.

Foregene ni imwe mu masosiyete akoresha tekinoroji ya PCR, kugirango amenye ORF1ab isanzwe, N, E, navariant imirongo ya acide nucleic acide ya nasofaryngeal ya oropharyngeal swab urugero nka SARS-CoV-2 B.1.1.7 umurongo (UK), B.1.351 umurongo (ZA), B.1.617 (IND) numurongo wa P.1 (BR).

02  Reagents ikenewe kuri PCR itaziguye

Icyitegererezo Lysate

Icyitegererezo lysate irashobora gushyirwaho wenyine cyangwa waguzwe.Itandukaniro mubigize ibirango bitandukanye bya lysate bizatuma ubushobozi bwa lysing butandukanye, hanyuma igihe cyo guterana kizaba gitandukanye gato.Kurugero, mugutegura ingero zinyama zinyamanswa, iminota 30 cyangwa ijoro ryose lysis irasabwa, kandi igisubizo cya lysis ya virusi kiri hagati yiminota 3-10.

PCR master mix

Birasabwa gukoresha ADN ishyushye-itangira polymerase kugirango yongere amplification kandi yongere ubushobozi bwo kongera imbaraga.Intangiriro ya PCR itaziguye ni polymerase yihanganira cyane.

Kuraho cyangwa kubuza ibice murugero bigira ingaruka kumyanya ya ADN

Icyitegererezo kimaze gutunganywa na lysate, proteyine, lipide nindi myanda ya selile irekurwa, ibyo bintu bizabuza PCR kwitwara.Kubwibyo, PCR itaziguye isaba kongerwaho gukuraho cyangwa kubuza kugabanya ingaruka zibi bintu.

03  Icyegeranyo cyibintu bitanu byubumenyi bya PCR itaziguye

Ubwa mbere, tekinoroji ya PCR ni tekinoroji ya PCR itaziguye kubinyabuzima bitandukanye.Muri ubu buryo bwa tekiniki, nta mpamvu yo gutandukanya no gukuramo aside nucleique, koresha mu buryo butaziguye icyitegererezo cya tissue nkikintu, hanyuma wongere intego ya gene primers ni ugukora reaction ya PCR.

Icya kabiri, tekinoroji ya PCR ntabwo ari tekinoroji gakondo ya ADN yerekana uburyo bwo kongera imbaraga, ahubwo ikubiyemo na RNA inyandikorugero ihinduranya PCR.

Icya gatatu, tekinoroji ya Directeur PCR ntabwo ikora gusa muburyo busanzwe bwa PCR ibisubizo byintangarugero, ahubwo inashyiramo reaction ya Real-Time qPCR, isaba sisitemu yogukora kugira ubushobozi bukomeye bwo kurwanya anti-background fluorescence hamwe na endogenous fluorescence ikuraho ubushobozi bwa antagonistique.

Icya kane, ibyitegererezo byibasiwe nubuhanga bwa Direct PCR bikenera gusa kurekura inyandikorugero ya acide nucleic, kandi ntibikuraho poroteyine, polysaccharide, ion yumunyu, nibindi bibangamira reaction ya PCR.Bikaba bisaba aside nucleique aside polymerase na PCR Ivanga muri sisitemu yo kubyitwaramo kugirango igire imbaraga zo guhangana noguhuza neza kugirango ibikorwa bya enzyme no kwigana neza mubihe bigoye.

Icya gatanu, icyitegererezo cya tissue cyerekanwe na tekinoroji ya PCR idafite uburyo bwo kuvura aside nucleique kandi ingano yicyitegererezo ni nto cyane, bisaba ko sisitemu yo kubyitwaramo iba ifite sensibilité nini kandi ikora neza.