PCR tooska ah waa falcelin si toos ah u adeegsata unugyada xayawaanka ama dhirta si ay u kordhiso iyada oo aan la soo saarin aashitada nucleic.Siyaabo badan, PCR tooska ah wuxuu u shaqeeyaa sida PCR caadiga ah

Farqiga ugu weyni waa kaydiyaha caadiga ah ee loo isticmaalo PCR tooska ah, muunada waxaa si toos ah loogu gudbin karaa falcelinta PCR iyada oo aan la soo saarin aashitada nukliyeerka, laakiin waxaa jira shuruudo u dhigma dulqaadka enzymes-ka iyo iswaafajinta kaydiyaha ku lug leh falcelinta tooska ah ee PCR.

Inkasta oo ay jiraan wax badan ama ka yar inhibitors PCR ee shaybaarada caadiga ah, PCR tooska ah ayaa weli ku guuleysan kara kor u qaadis la isku halayn karo iyada oo la raacayo ficilka enzymes iyo buffers.Falcelinta PCR-da dhaqameed waxay u baahan tahay aashito nucleic ah oo tayo sare leh qaab template ah, taas oo hor istaagi karta horumarka siman ee falcelinta PCR haddii qaabku ka kooban yahay borotiinno iyo wasakh kale.PCR tooska ah hadda waa mid ka mid ah tignoolajiyada caanka ah ee goobta baadhista molecular.

01 Direct PCR waxaa markii hore loo isticmaali jiray xoolaha iyo dhirta

Codsiga ugu horreeya ee PCR tooska ah waa duurka xoolaha iyo dhirta, sida dhiigga, unugyada iyo timaha jiirka, bisad, digaag, bakayle, idaha, lo'da, iwm, caleemaha dhirta iyo miraha, iwm, loo isticmaalo in lagu barto genotyping, transgenic, ogaanshaha plasmid, Falanqaynta Gene knockout, aqoonsiga isha DNA, aqoonsiga noocyada, falanqaynta SNP iyo beeraha kale.

Goobahani waxay leeyihiin qaar ka mid ah sifooyin caadi ah, taas oo ah, nuxurka hidde-sidaha bartilmaameedku waa mid aad u sarreeya iyo soo saarista nucleic acid waa dhib badan, markaa PCR tooska ah ma badbaadin karto oo kaliya waqti iyo saameyn yar oo natiijooyinka, laakiin sidoo kale waxay badbaadisaa kharashka.

PCR-ga tooska ah ee loo isticmaalo ogaanshaha cudur-sidaha waa arrin sannadihii la soo dhaafay, qaar ka mid ah soo-saareyaasha PCR reagent waxay sameeyeen dadaallo badan oo dhinacan ah markii ay samaynayeen hal-abuurnimo.Gaar ahaan cudurkan faafa ee COVID-19, alaabooyin badan oo noocaan oo kale ah ayaa ka soo muuqday suuqa, sida Qalabka Ogaanshaha Acid Nucleic Acid (SARS-CoV-2 Nucleic Acid Detection Kit) oo ay soo saartay Foregene, taas oo u adeegsata tikniyoolajiyadda RT PCR ee waqtiga-dhabta ah (rRT-PCR) si loo ogaado tayada SARS-CoV-2 ee muunadda nukleic acids ee bini'aadamka.

Foregene waa mid ka mid ah shirkadaha isticmaala tignoolajiyada tooska ah ee PCR, si loo ogaado ORF1ab, N, E, iyokala duwanaansho lineages nucleic acids ee nasopharyngeal bini'aadamka ama muunado swab oropharyngeal sida SARS-CoV-2 B.1.1.7 lineage (UK), B.1.351 lineage (ZA), B.1.617 lineage (IND) iyo P.1 lineage (BR).

02  Reagents ayaa looga baahan yahay PCR tooska ah

Tusaalaha Lysate

Muunada lysate-ka waxaa habayn kara naftaada ama waa la iibsan karaa.Farqiga u dhexeeya noocyada kala duwan ee lysate ayaa ka dhigi doona awoodda been-abuurka ah ee kala duwan, ka dibna wakhtiga been-abuurka ayaa wax yar ka duwanaan doona.Tusaale ahaan, diyaarinta shaybaarka unugyada xoolaha, 30 daqiiqo ama lysis habeen habeen ah ayaa guud ahaan lagula talinayaa, iyo xalka lysis ee fayraska wuxuu u dhexeeyaa 3-10 daqiiqo.

PCR Master mix

Waxaa lagu talinayaa in la isticmaalo polymerase DNA-ga bilawga ah si kor loogu qaado kor u qaadis gaar ah oo loo kordhiyo awoodda weyneynta.Xudunta PCR tooska ah waa polymerase dulqaad badan.

Meesha ka saar ama jooji qaybaha muunada ee saameeya waynaynta DNA-da

Ka dib marka muunada lagu farsameeyo lysate, borotiinada, lipids iyo haraaga unugyada kale ayaa la sii deyn doonaa, walxahan ayaa joojin doona falcelinta PCR.Sidaa darteed, PCR tooska ah waxay u baahan tahay in lagu daro ka saarista u dhiganta ama horjoogayaasha si loo yareeyo saameynta arrimahan.

03  Ururinta shan dhibcood oo aqooneed PCR tooska ah

Marka hore, tignoolajiyada tooska ah ee PCR waa tignoolajiyada PCR ee tooska ah ee muunado noole oo kala duwan.Marka la eego xaaladdan farsamo, looma baahna in la kala saaro oo la soo saaro aashitada nucleic, si toos ah u isticmaal muunadda nudaha shayga, oo lagu daro bartilmaameedyada hidda-wadaha si loo sameeyo falcelinta PCR.

Midda labaad, tignoolajiyada tooska ah ee PCR kaliya maahan tignoolajiyada kobcinta template DNA ee dhaqameed, laakiin sidoo kale waxaa ku jira template RNA PCR-ga beddelka qoraalka.

Seddexaad, tignoolajiyada tooska ah ee PCR ma aha oo kaliya inay si toos ah u fuliso falcelinta PCR ee tayada joogtada ah ee shaybaarada unugyada, laakiin sidoo kale waxaa ku jira falcelinta qPCR-waqtiga-dhabta ah, taas oo u baahan nidaamka falcelinta si uu u yeesho awood faragelineed oo ka hortag ah oo ka soo horjeeda asalka iyo fluorescence endogenous demiya awooda iska soo horjeeda.

Midda afraad, shaybaarrada lagu bartilmaameedsaday tignoolajiyada tooska ah ee PCR waxay u baahan yihiin oo keliya in la sii daayo habyaalada aashitada nukliyeerka, oo ha ka saarin borotiinnada, polysaccharides, ions milixda, iwm ee farageliya falcelinta PCR.Taas oo u baahan polymerase nucleic acid iyo PCR Isku darka habka falcelinta si ay u yeeshaan iska caabin heer sare ah iyo la qabsi si loo hubiyo dhaqdhaqaaqa enzyme iyo saxnaanta ku celcelinta xaaladaha adag.

Shanaad, muunadda nudaha ay bartilmaameedsatay tignoolajiyada tooska ah ee PCR iyada oo aan la helin daawaynta kobcinta aashitada nukliyeerka iyo cadadka template waa mid aad u yar, kaas oo u baahan nidaamka falcelinta si uu u yeesho dareen aad u sarreeya iyo waxtarka xoojinta.