{"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\/nb\/rt-pcr-test-for-koronavirus\/","title":{"rendered":"RT-PCR-test for koronaviruset"},"content":{"rendered":"

Av alt du har h\u00f8rt om covid-19 i 2020, er det \u00e9n ting som er midt i blinken: testen som brukes til \u00e5 p\u00e5vise om noen er smittet med viruset, den ber\u00f8mte RT-PCR-testen eller bare PCR.<\/span><\/p>\n

Surely, you have already heard about this test before, from a medical professional or in the news. PCR test is employed to detect the presence (or not) of the virus in someone’s body.<\/span><\/p>\n

RT-PCR-testen er ikke den eneste som kan p\u00e5vise viruset, men sammenlignet med de andre metodene er det den mest n\u00f8yaktige og p\u00e5litelige. En annen test er den konvensjonelle serologitesten som p\u00e5viser spesifikke antistoffniv\u00e5er i blodpr\u00f8ver.<\/span><\/p>\n

Bokstavene i RT-PCR st\u00e5r for <\/span>Polymerasekjedereaksjon med omvendt transkripsjon<\/b>I flere ti\u00e5r har den blitt brukt av forskere til \u00e5 produsere flere kopier av genetisk materiale.<\/span><\/p>\n

Med andre ord, i dette tilfellet, mer genetisk materiale fra SARS-CoV-2, som brukes som fingeravtrykk for \u00e5 p\u00e5vise tilstedev\u00e6relsen av viruset i pr\u00f8ver fra nese, \u00f8rer eller spytt fra pasienter. Hvis det finnes virus skjult i pr\u00f8ven, selv i sm\u00e5 mengder, vil det kunne p\u00e5vises p\u00e5 grunn av kopi-lim-syklusen. Det er derfor RT-PCR-testen er s\u00e5 p\u00e5litelig.<\/span><\/p>\n

Testen<\/b> er en metode med i utgangspunktet to trinn i temperatursyklusen, der man kopierer og limer inn et stykke genetisk materiale, enten det er DNA eller RNA.<\/span><\/p>\n

Testen krever en <\/span>termosykler-maskin<\/b> som har en presis <\/span>temperatur- og tidsstyring<\/b>. Gjennom flere temperatursykluser vil virusets genetiske materiale \u00f8ke. Nedenfor vises et eksempel p\u00e5 en temperatursyklus.<\/span><\/p>\n

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The method requires two main enzymes, the Reverse Transcriptase (that\u2019s why the \u201cRT\u201d in the title) and the DNA-polymerase. Each one of them has a phase of action and a function. In the figure above, the yellow phase of the cycle is when the RT acts, and the red phase is when DNA-polymerase does its job.<\/span><\/p>\n

Det f\u00f8rste enzymet lager den komplement\u00e6re DNA-strengen fra RNA-viruset, slik at det andre enzymet kan utf\u00f8re arbeidet med \u00e5 kopiere og lime inn den nye DNA-strengen, og dermed \u00f8ke mengden genetisk materiale. Denne amplifikasjonen gj\u00f8r det mulig \u00e5 karakterisere og\/eller manipulere det genetiske materialet, noe som ikke ville v\u00e6rt mulig i en mindre mengde.<\/span><\/p>\n

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For all of this process occur the enzymes need help from a small (or big) molecule called PRIMER. The primer function is to indicate the location where the enzyme should begin to extend the strands, without it the copy-paste process of the DNA-polymerase won’t start.<\/span><\/p>\n

Primeren kan velges med en tilfeldig eller en spesifikk form. I slike tilfeller er det sv\u00e6rt viktig \u00e5 kjenne det genetiske kartet til v\u00edrus, slik at man kan bruke spesifikke primere for \u00e5 \u00f8ke effektiviteten og p\u00e5liteligheten til resultatene.<\/span><\/p>\n

Hvis du er like nysgjerrig som oss og vil vite mer om RT-PCR, kan du lese mer om dette her.\u00a0<\/span> denne vitenskapelige artikkelen <\/span>her<\/span><\/a>\u00a0 men her er i hovedsak alt som trengs for \u00e5 utf\u00f8re testen:<\/span><\/p>\n

Virusets genetiske materiale, RNA<\/strong><\/span><\/p>\n

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

Omvendt transkriptase og DNA-polymerase-enzymer<\/b><\/span><\/p>\n

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

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

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

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

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

og fluorescerende rapporteringsfargestoffer:\u00a0<\/b><\/span><\/p>\n

For \u00e5 vite resultatene av en pr\u00f8ve som kan eller ikke kan v\u00e6re forurenset med COVID-19-virus, brukes fluorescerende reporterfargestoffer i kombinasjon med RT-PCR-testen. Analysen er basert p\u00e5 tilstedev\u00e6relsen eller frav\u00e6ret av et fluorescerende signal i en pr\u00f8ve. I frav\u00e6r av et signal kan testen konkluderes som negativ eller falsk negativ. Ved tilstedev\u00e6relse av et intenst signal kan det ikke bare konkluderes med at pr\u00f8ven er positiv for tilstedev\u00e6relse av v\u00edrus, men ogs\u00e5 at sykdommen er i et avansert stadium.<\/span><\/p>\n

Bland alle ingrediensene og legg dem i termosyklusmaskinen.\u00a0<\/b><\/span><\/p>\n

Det f\u00f8rste trinnet i testen vises i infografikken nedenfor. RT-enzymet lager den f\u00f8rste kopien av virusets genetiske materiale i pr\u00f8ven. Deretter f\u00f8lger det andre trinnet i RT-PCR-metoden. Primeren angir startposisjonen, og DNA-polymerasen begynner \u00e5 forlenge DNA-strengene ved hjelp av en enzymatisk reaksjon.<\/span><\/p>\n

\"RT-PCR-test<\/a><\/p>\n

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

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

Finally, this picture is an electron micrograph image of the SARS-CoV-2 virus, and it is possible to see perfectly the structure of the virus. Isn’t that beautiful?\u00a0\u00a0<\/span><\/p>\n

Testen er kompleks og ikke s\u00e5 enkel som den ser ut til \u00e5 v\u00e6re, men med infografikken var det sikkert mye lettere \u00e5 forst\u00e5 helheten i hvordan den fungerer, ikke sant?<\/span><\/p>\n

Det kan du gj\u00f8re med hvilken som helst vitenskapelig informasjon du \u00f8nsker \u00e5 bruke.\u00a0<\/span> Mind the Graph<\/span><\/a>. Plattformen er her for \u00e5 gi deg frihet til \u00e5 kommunisere vitenskapelig informasjon p\u00e5 en bedre m\u00e5te. L\u00e6r mer om hvordan du bruker plattformen, og kontakt oss hvis du trenger hjelp.<\/span>!<\/b><\/p>\n

Referanse<\/b><\/p>\n

AI, T. et al. Correlation of Chest CT and RT-PCR Testing for Coronavirus Disease 2019 (COVID-19) in China: A Report of 1014 Cases. Radiology, v. 296, n. 2, p. E32\u2013E40, 26 fev. 2020.<\/span><\/p>\n

GIBELLINI, D. et al. Kvantitativ p\u00e5visning av humant immunsviktvirus type 1 (HIV-1) viral belastning ved SYBR gr\u00f8nn sanntids RT-PCR-teknikk hos HIV-1 seropositive pasienter. Journal of Virological Methods, v. 115, n. 2, s. 183-189, 1 fev. 2004.<\/span><\/p>\n

JAMES, D. En enkel og p\u00e5litelig protokoll for p\u00e5visning av eple stamme grooving virus ved RT-PCR og i en multiplex PCR-analyse. Journal of Virological Methods, v. 83, n. 1, s. 1-9, 1 dez. 1999.<\/span><\/p>\n

LI, Y. et al. Stabilitetsproblemer ved RT-PCR-testing av SARS-CoV-2 for innlagte pasienter som er klinisk diagnostisert med COVID-19. Journal of Medical Virology, v. 92, nr. 7, s. 903-908, 2020.<\/span><\/p>\n

LIAO, C.-S. et al. Miniature RT\u2013PCR system for diagnosis of RNA-based viruses. Nucleic Acids Research, v. 33, n. 18, p. e156\u2013e156, 1 out. 2005.<\/span><\/p>\n

MOHAN, A. et al. Prevalence of viral infection detected by PCR and RT-PCR in patients with acute exacerbation of COPD: A systematic review. Respirology, v. 15, n. 3, p. 536\u2013542, 2010.<\/span><\/p>\n

NOLAN, T.; HANDS, R. E.; BUSTIN, S. A. Quantification of mRNA using real-time RT-PCR. Nature Protocols, v. 1, n. 3, p. 1559\u20131582, ago. 2006.<\/span><\/p>\n

Det nye koronaviruset SARS-CoV-2 | Dette overf\u00f8ringselektronm... | Flickr. Tilgjengelig p\u00e5: . Tilgjengelig fra: 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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He has a Ph.D. and solid scientific background in Psychopharmacology and experience as a Guest Researcher at the Max Planck Institute of Psychiatry (Germany) and Researcher in D'Or Institute for Research and Education (IDOR, Brazil). Fabricio holds over 2500 citations in Google Scholar. He has 10 years of experience in small innovative businesses, with relevant experience in product design and innovation management. 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