{"id":11803,"date":"2020-10-28T09:30:35","date_gmt":"2020-10-28T11:30:35","guid":{"rendered":"https:\/\/mindthegraph.com\/blog\/?p=11803"},"modified":"2022-10-18T08:09:32","modified_gmt":"2022-10-18T11:09:32","slug":"rt-pcr-test-for-coronavirus","status":"publish","type":"post","link":"https:\/\/mindthegraph.com\/blog\/sl\/rt-pcr-test-za-koronavirus\/","title":{"rendered":"Test RT-PCR za koronavirus"},"content":{"rendered":"

Iz \u0161tevilnih stvari, ki ste jih sli\u0161ali o COVID-19 v letu 2020, je sredi vsega tega test, ki se uporablja za ugotavljanje, ali je nekdo oku\u017een z virusom, znani test RT-PCR ali samo PCR.<\/span><\/p>\n

Zagotovo ste za ta test \u017ee sli\u0161ali od zdravnika ali v novicah. Test PCR se uporablja za ugotavljanje prisotnosti (ali odsotnosti) virusa v telesu.<\/span><\/p>\n

Test RT-PCR ni edini, s katerim je mogo\u010de odkriti virus, vendar je v primerjavi z drugimi metodami najbolj natan\u010den in zanesljiv. Drug test je obi\u010dajni serolo\u0161ki test, s katerim v vzorcih krvi odkrijemo specifi\u010dne ravni protiteles.<\/span><\/p>\n

\u010crke v RT-PCR pomenijo <\/span>Reverzna transkripcijska veri\u017ena reakcija s polimerazo<\/b>, ga znanstveniki \u017ee desetletja uporabljajo za izdelavo ve\u010d kopij genskega materiala.<\/span><\/p>\n

Z drugimi besedami, v tem primeru je ve\u010d genetskega materiala SARS-CoV-2, ki se uporablja kot prstni odtis za dokazovanje prisotnosti virusa v vzorcih iz nosu, u\u0161es ali sline, zbranih pri bolnikih. \u010ce so v vzorcu skriti kakr\u0161ni koli virusi, \u010detudi v majhnih koli\u010dinah, jih bo lahko odkril zaradi svojega cikla kopiranja in prilepljanja. Zato je test RT-PCR tako zanesljiv.<\/span><\/p>\n

Test<\/b> je metoda z dvema korakoma znotraj temperaturnega cikla, ki v bistvu izvaja postopek kopiranja in prilepljanja dela genskega materiala, bodisi DNK ali RNK.<\/span><\/p>\n

Test zahteva <\/span>termocikli\u010dni stroj<\/b> ki ima natan\u010den <\/span>nadzor temperature in \u010dasa<\/b>. Z ve\u010d temperaturnimi cikli se virusni genski material pove\u010da. Primer temperaturnega cikla je prikazan v nadaljevanju.<\/span><\/p>\n

\"\"<\/a><\/p>\n

Metoda zahteva dva glavna encima, reverzno transkriptazo (zato je v naslovu \"RT\") in DNK-polimerazo. Vsak od njiju ima svojo fazo delovanja in funkcijo. Na zgornji sliki je rumena faza cikla, ko deluje RT, in rde\u010da faza, ko DNK-polimeraza opravlja svoje delo.<\/span><\/p>\n

Prvi encim izdela komplementarno verigo DNK iz virusa RNK, kar omogo\u010da drugemu encimu, da opravi delo kopiranja in prilepitve iz nove, pravkar izdelane verige DNK, s \u010dimer se pove\u010da koli\u010dina prisotnega genskega materiala. To pomno\u017eevanje omogo\u010da karakterizacijo in\/ali manipulacijo genetskega materiala, ki v manj\u0161i koli\u010dini ne bi bila mogo\u010da.<\/span><\/p>\n

\"\"<\/a><\/p>\n

Pri vseh teh procesih encimi potrebujejo pomo\u010d majhne (ali velike) molekule, imenovane PRIMER. Naloga primerja je, da ozna\u010di mesto, kjer naj encim za\u010dne podalj\u0161evati verige, brez njega se postopek kopiranja in lepljenja DNK-polimeraze ne bo za\u010del.<\/span><\/p>\n

Primer lahko izberete v naklju\u010dni ali dolo\u010deni obliki. V tak\u0161nih primerih je zelo pomembno poznati genetski zemljevid virusa, da se lahko uporabijo posebni primeri za pove\u010danje u\u010dinkovitosti in zanesljivosti rezultatov.<\/span><\/p>\n

\u010ce ste tako kot mi radovedni in \u017eelite izvedeti ve\u010d podrobnosti o RT-PCR, lahko preverite ve\u010d podrobnosti\u00a0<\/span> ta znanstveni \u010dlanek <\/span>tukaj<\/span><\/a>\u00a0 v bistvu pa je tukaj vse, kar je potrebno za izvedbo testa:<\/span><\/p>\n

Virusni genetski material, RNK<\/strong><\/span><\/p>\n

\"\"<\/a><\/p>\n

Encimi reverzne transkriptaze in DNK-polimeraze<\/b><\/span><\/p>\n

\"\"<\/a><\/p>\n

PRIMERJI<\/b><\/span><\/p>\n

\"\"<\/a><\/p>\n

Nukleozidni trifosfati<\/b><\/span><\/p>\n

\"\"<\/a><\/p>\n

In fluorescen\u010dna reporterska barvila:\u00a0<\/b><\/span><\/p>\n

Za ugotavljanje rezultatov vzorca, ki je morda oku\u017een z virusom COVID-19 ali ne, se v kombinaciji s testom RT-PCR uporablja test s fluorescen\u010dnimi reporterskimi barvili. Test temelji na prisotnosti ali odsotnosti fluorescen\u010dnega signala vzorca. \u010ce signala ni, je test lahko negativen ali la\u017eno negativen. \u010ce je prisoten intenziven signal, je lahko test pozitiven ne le za prisotnost virusa, temve\u010d tudi za napredovalo stopnjo bolezni.<\/span><\/p>\n

Zme\u0161ajte vse sestavine in jih dajte v termocikel.\u00a0<\/b><\/span><\/p>\n

Prvi korak testa je prikazan v naslednji infografiki. Encim RT naredi prvo kopijo genetskega materiala virusa, ki je prisoten v vzorcu. Sledi drugi korak metode RT-PCR. Primer ozna\u010di za\u010detni polo\u017eaj in DNK-polimeraza z encimsko reakcijo za\u010dne podalj\u0161evati verige DNK.<\/span><\/p>\n

\"Test<\/a><\/p>\n

\"\"<\/a><\/p>\n

\"\"<\/a><\/p>\n

Na tej sliki je elektronska mikrografija virusa SARS-CoV-2, na kateri je dobro vidna struktura virusa. Ali ni to \u010dudovito?\u00a0\u00a0<\/span><\/p>\n

Test je zapleten in ni tako preprost, kot se zdi, vendar stavim, da je bilo z infografikami veliko la\u017eje razumeti celotno sliko njegovega delovanja, kajne?<\/span><\/p>\n

To lahko storite z vsemi znanstvenimi podatki, ki jih \u017eelite, z uporabo\u00a0<\/span> Mind the Graph<\/span><\/a>. Platforma je tu zato, da vam omogo\u010di avtonomijo pri bolj\u0161em posredovanju znanstvenih informacij. Preberite ve\u010d o tem, kako uporabljati platformo, in \u010de potrebujete pomo\u010d, se obrnite na nas.<\/span>!<\/b><\/p>\n

Referenca<\/b><\/p>\n

AI, T. et al. Korelacija CT prsnega ko\u0161a in testiranja RT-PCR za koronavirusno bolezen 2019 (COVID-19) na Kitajskem: Poro\u010dilo o 1014 primerih. Radiology, v. 296, \u0161t. 2, str. E32-E40, 26. feb. 2020.<\/span><\/p>\n

GIBELLINI, D. et al. Kvantitativno odkrivanje virusne obremenitve virusa \u010dlove\u0161ke imunske pomanjkljivosti tipa 1 (HIV-1) s tehniko SYBR green RT-PCR v realnem \u010dasu pri seropozitivnih bolnikih s HIV-1. Journal of Virological Methods, v. 115, \u0161t. 2, str. 183-189, 1. februar 2004.<\/span><\/p>\n

JAMES, D. Preprost in zanesljiv protokol za odkrivanje virusa groovinga jablanovega peclja z RT-PCR in s testom multiplex PCR. Journal of Virological Methods, v. 83, \u0161t. 1, str. 1-9, 1. dez. 1999.<\/span><\/p>\n

LI, Y. et al. Vpra\u0161anja stabilnosti testiranja RT-PCR SARS-CoV-2 pri hospitaliziranih bolnikih s klini\u010dno diagnozo COVID-19. Journal of Medical Virology, v. 92, \u0161t. 7, str. 903-908, 2020.<\/span><\/p>\n

LIAO, C.-S. et al. Miniaturni sistem RT-PCR za diagnosticiranje virusov na osnovi RNK. Nucleic Acids Research, v. 33, \u0161t. 18, str. e156-e156, 1 izid. 2005.<\/span><\/p>\n

MOHAN, A. et al. Raz\u0161irjenost virusnih oku\u017eb, ugotovljenih s PCR in RT-PCR, pri bolnikih z akutnim poslab\u0161anjem KOPB: sistemati\u010dni pregled. Respirology, letnik 15, \u0161t. 3, str. 536-542, 2010.<\/span><\/p>\n

NOLAN, T.; HANDS, R. E.; BUSTIN, S. A. Kvantifikacija mRNA z RT-PCR v realnem \u010dasu. Nature Protocols, v. 1, \u0161t. 3, str. 1559-1582, nazaj. 2006.<\/span><\/p>\n

Novi koronavirus SARS-CoV-2 | Ta transmisijski elektronski mi... | Flickr. Na voljo v: . Dostopno prek: 23 jul. 2020. <\/span><\/p>","protected":false},"excerpt":{"rendered":"","protected":false},"author":4,"featured_media":11847,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[959,28],"tags":[388,799,554,798,250],"yoast_head":"\nThe RT-PCR Test for Coronavirus - Mind the Graph Blog<\/title>\n<meta name=\"description\" content=\"Read this blog to know everything related to the popular COVID-19 test, RT-PCR or only PCR. 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