{"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\/da\/rt-pcr-test-for-coronavirus\/","title":{"rendered":"RT-PCR-test for coronavirus"},"content":{"rendered":"

Af de mange ting, du har h\u00f8rt om COVID-19 i 2020, er der \u00e9n ting midt i det hele, nemlig den test, der bruges til at opdage, om nogen er smittet med virussen, den ber\u00f8mte RT-PCR-test eller bare PCR.<\/span><\/p>\n

Du har helt sikkert allerede h\u00f8rt om denne test f\u00f8r, fra en l\u00e6ge eller i nyhederne. PCR-testen bruges til at p\u00e5vise tilstedev\u00e6relsen (eller ej) af virussen i en persons krop.<\/span><\/p>\n

RT-PCR-testen er ikke den eneste, der kan p\u00e5vise virussen, men sammenlignet med de andre metoder er det den mest n\u00f8jagtige og p\u00e5lidelige. En anden test er den konventionelle serologiske test, der p\u00e5viser specifikke antistofniveauer i blodpr\u00f8ver.<\/span><\/p>\n

Bogstaverne i RT-PCR st\u00e5r for <\/span>Polymerase-k\u00e6dereaktion med omvendt transkription<\/b>Det har v\u00e6ret brugt i \u00e5rtier af forskere til at producere flere kopier af genetisk materiale.<\/span><\/p>\n

Med andre ord, i dette tilf\u00e6lde, mere genetisk materiale fra SARS-CoV-2, som bruges som fingeraftryk til at bevidne tilstedev\u00e6relsen af virussen i pr\u00f8ver fra n\u00e6se, \u00f8rer eller spyt indsamlet fra patienter. Hvis der er virus gemt i pr\u00f8ven, selv i sm\u00e5 m\u00e6ngder, vil det kunne detekteres p\u00e5 grund af dets copy-paste-cyklus. Det er derfor, RT-PCR-testen er s\u00e5 p\u00e5lidelig.<\/span><\/p>\n

Testen<\/b> er en metode med grundl\u00e6ggende to trin i en temperaturcyklus, som udf\u00f8rer en copy-paste-proces af et stykke genetisk materiale, hvad enten det er DNA eller RNA.<\/span><\/p>\n

Testen kr\u00e6ver en <\/span>termocykler-maskine<\/b> som har en pr\u00e6cis <\/span>temperatur- og tidsstyring<\/b>. Gennem flere temperaturcyklusser vil det virale genetiske materiale \u00f8ges. Et eksempel p\u00e5 en temperaturcyklus er vist nedenfor.<\/span><\/p>\n

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

Metoden kr\u00e6ver to hovedenzymer, Reverse Transcriptase (det er derfor, der st\u00e5r \"RT\" i titlen) og DNA-polymerase. Hver af dem har en virkningsfase og en funktion. I figuren ovenfor er den gule fase af cyklussen, n\u00e5r RT virker, og den r\u00f8de fase er, n\u00e5r DNA-polymerase g\u00f8r sit arbejde.<\/span><\/p>\n

Det f\u00f8rste enzym laver den komplement\u00e6re DNA-streng fra RNA-virussen, s\u00e5 det andet enzym kan udf\u00f8re arbejdet med at kopiere og inds\u00e6tte fra den nye DNA-streng, der lige er lavet, hvilket \u00f8ger m\u00e6ngden af genetisk materiale. Denne amplifikation g\u00f8r det muligt at karakterisere og\/eller manipulere det genetiske materiale, hvor det ikke ville v\u00e6re muligt i en mindre m\u00e6ngde.<\/span><\/p>\n

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

For at alle disse processer kan finde sted, har enzymerne brug for hj\u00e6lp fra et lille (eller stort) molekyle kaldet PRIMER. Primerens funktion er at angive det sted, hvor enzymet skal begynde at forl\u00e6nge strengene, uden den vil DNA-polymerasens copy-paste-proces ikke starte.<\/span><\/p>\n

Primeren kan v\u00e6lges med en tilf\u00e6ldig eller en specifik form. Det er meget vigtigt at kende det genetiske kort over v\u00edrus i tilf\u00e6lde som dette, s\u00e5 man kan bruge specifikke primere til at \u00f8ge effektiviteten og p\u00e5lideligheden af resultaterne.<\/span><\/p>\n

Hvis du ligesom os er nysgerrig og gerne vil vide mere om RT-PCR, kan du l\u00e6se mere her.\u00a0<\/span> denne videnskabelige artikel <\/span>lige her<\/span><\/a>\u00a0 men i bund og grund er her alt, hvad der er n\u00f8dvendigt for at udf\u00f8re testen:<\/span><\/p>\n

Virus' genetiske materiale, RNA'et<\/strong><\/span><\/p>\n

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

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

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

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

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

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

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

Og fluorescerende reporterfarvestoffer:\u00a0<\/b><\/span><\/p>\n

For at kende resultaterne af en pr\u00f8ve, der m\u00e5ske eller m\u00e5ske ikke er kontamineret med COVID-19-virus, bruges fluorescerende reporterfarvestoffer i kombination med RT-PCR-testen. Assayet er baseret p\u00e5 tilstedev\u00e6relsen eller frav\u00e6ret af et fluorescerende signal i pr\u00f8ven. Hvis der ikke er et signal, kan testen konkluderes som negativ eller falsk-negativ. Hvis der er et intenst signal, kan det konkluderes, at det ikke kun er positivt for tilstedev\u00e6relsen af v\u00edrus, men ogs\u00e5 som et fremskredent stadie af sygdommen.<\/span><\/p>\n

Bland alle ingredienserne, og kom det i termocyklusmaskinen.\u00a0<\/b><\/span><\/p>\n

Testens f\u00f8rste trin er vist i den n\u00e6ste infografik. RT-enzymet laver den f\u00f8rste kopi af det genetiske materiale fra den virus, der findes i pr\u00f8ven. Herefter ser du det andet trin i RT-PCR-metoden. Primeren angiver startpositionen, og DNA-polymerasen begynder gennem en enzymatisk reaktion at forl\u00e6nge DNA-strengene.<\/span><\/p>\n

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

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

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

Endelig er dette billede et elektronmikrografisk billede af SARS-CoV-2-virusset, og det er muligt at se virussets struktur perfekt. Er det ikke smukt?\u00a0\u00a0<\/span><\/p>\n

Testen er kompleks og ikke s\u00e5 enkel, som den ser ud til at v\u00e6re, men med infografikken var det garanteret meget lettere at forst\u00e5 det store billede af, hvordan den fungerer, ikke?<\/span><\/p>\n

Det kan du g\u00f8re med al den videnskabelige information, du \u00f8nsker, ved hj\u00e6lp af\u00a0<\/span> Mind the Graph<\/span><\/a>. Platformen er her for at give dig autonomi til bedre at kommunikere videnskabelig information. F\u00e5 mere at vide om, hvordan du bruger platformen, og kontakt os, hvis du har brug for hj\u00e6lp.<\/span>!<\/b><\/p>\n

Reference<\/b><\/p>\n

AI, T. et al. Korrelation mellem bryst-CT og RT-PCR-test for Coronavirus Disease 2019 (COVID-19) i Kina: En rapport om 1014 tilf\u00e6lde. Radiology, v. 296, n. 2, p. E32-E40, 26 fev. 2020.<\/span><\/p>\n

GIBELLINI, D. et al. Kvantitativ p\u00e5visning af human immundefektvirus type 1 (HIV-1) viral belastning med SYBR green real-time RT-PCR-teknik hos HIV-1 seropositive patienter. Journal of Virological Methods, v. 115, n. 2, s. 183-189, 1 fev. 2004.<\/span><\/p>\n

JAMES, D. A simple and reliable protocol for the detection of apple stem grooving virus by RT-PCR and in a multiplex PCR assay. 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-test af SARS-CoV-2 for indlagte patienter, der er klinisk diagnosticeret med COVID-19. Journal of Medical Virology, v. 92, n. 7, s. 903-908, 2020.<\/span><\/p>\n

LIAO, C.-S. et al. Miniature RT-PCR-system til diagnosticering af RNA-baserede vira. Nucleic Acids Research, v. 33, n. 18, p. e156-e156, 1 out. 2005.<\/span><\/p>\n

MOHAN, A. et al. Pr\u00e6valens af virusinfektion p\u00e5vist ved PCR og RT-PCR hos patienter med akut forv\u00e6rring af KOL: En systematisk gennemgang. Respirology, v. 15, n. 3, s. 536-542, 2010.<\/span><\/p>\n

NOLAN, T.; HANDS, R. E.; BUSTIN, S. A. Kvantificering af mRNA ved hj\u00e6lp af RT-PCR i realtid. Nature Protocols, v. 1, n. 3, s. 1559-1582, ago. 2006.<\/span><\/p>\n

Nyt coronavirus SARS-CoV-2 | Dette transmissionselektronbillede | Flickr. Dispon\u00edvel em: . Adgang til: 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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