{"id":16525,"date":"2021-03-02T00:00:00","date_gmt":"2021-03-01T23:00:00","guid":{"rendered":"https:\/\/www.ecoceutics.com\/funcionament-pcr\/"},"modified":"2023-04-11T13:11:17","modified_gmt":"2023-04-11T11:11:17","slug":"funcionament-pcr","status":"publish","type":"post","link":"https:\/\/www.ecoceutics.com\/ca\/respostes-de-salut\/salut\/funcionament-pcr\/","title":{"rendered":"En qu\u00e8 consisteix una PCR?"},"content":{"rendered":"<p>La reacci\u00f3 en cadena de la polimerasa (la famosa PCR) \u00e9s una t\u00e8cnica que s\u2019utilitza per a \u00abamplificar\u00bb petites quantitats d\u2019ADN. La sigla PCR ve del nom en angl\u00e8s de la t\u00e8cnica:&nbsp;Polymerase&nbsp;Chain&nbsp;Reaction. El material gen\u00e8tic en petita quantitat resulta invisible per als m\u00e8todes d\u2019an\u00e0lisis, fins i tot els m\u00e9s moderns, per la qual cosa \u00e9s necessari realitzar c\u00f2pies, moltes, moltes c\u00f2pies (d\u2019aqu\u00ed el verb \u00abamplificar\u00bb) per a poder estudiar-ho o detectar-ho. T\u2019expliquem una mica m\u00e9s detalladament aquesta t\u00e8cnica, tan trencadora que va rebre el premi Nobel de qu\u00edmica de l\u2019any 1993.<\/p>\n<div id=\"ez-toc-container\" class=\"ez-toc-v2_0_82_2 counter-hierarchy ez-toc-counter ez-toc-custom ez-toc-container-direction\">\n<div class=\"ez-toc-title-container\">\n<p class=\"ez-toc-title\" style=\"cursor:inherit\">Contenido de la p\u00e1gina<\/p>\n<span class=\"ez-toc-title-toggle\"><a href=\"#\" class=\"ez-toc-pull-right ez-toc-btn ez-toc-btn-xs ez-toc-btn-default ez-toc-toggle\" aria-label=\"Toggle Table of Content\"><span class=\"ez-toc-js-icon-con\"><span class=\"\"><span class=\"eztoc-hide\" style=\"display:none;\">Toggle<\/span><span class=\"ez-toc-icon-toggle-span\"><svg style=\"fill: #333333;color:#333333\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" class=\"list-377408\" width=\"20px\" height=\"20px\" viewBox=\"0 0 24 24\" fill=\"none\"><path d=\"M6 6H4v2h2V6zm14 0H8v2h12V6zM4 11h2v2H4v-2zm16 0H8v2h12v-2zM4 16h2v2H4v-2zm16 0H8v2h12v-2z\" fill=\"currentColor\"><\/path><\/svg><svg style=\"fill: #333333;color:#333333\" class=\"arrow-unsorted-368013\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"10px\" height=\"10px\" viewBox=\"0 0 24 24\" version=\"1.2\" baseProfile=\"tiny\"><path d=\"M18.2 9.3l-6.2-6.3-6.2 6.3c-.2.2-.3.4-.3.7s.1.5.3.7c.2.2.4.3.7.3h11c.3 0 .5-.1.7-.3.2-.2.3-.5.3-.7s-.1-.5-.3-.7zM5.8 14.7l6.2 6.3 6.2-6.3c.2-.2.3-.5.3-.7s-.1-.5-.3-.7c-.2-.2-.4-.3-.7-.3h-11c-.3 0-.5.1-.7.3-.2.2-.3.5-.3.7s.1.5.3.7z\"\/><\/svg><\/span><\/span><\/span><\/a><\/span><\/div>\n<nav><ul class='ez-toc-list ez-toc-list-level-1 ' ><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-1\" href=\"https:\/\/www.ecoceutics.com\/ca\/respostes-de-salut\/salut\/funcionament-pcr\/#que-es-la-reaccio-en-cadena-de-la-polimerasa\" >Qu\u00e8 \u00e9s la reacci\u00f3 en cadena de la polimerasa?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-2\" href=\"https:\/\/www.ecoceutics.com\/ca\/respostes-de-salut\/salut\/funcionament-pcr\/#com-funciona-la-reaccio-en-cadena-de-la-polimerasa\" >Com funciona la reacci\u00f3 en cadena de la polimerasa?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-3\" href=\"https:\/\/www.ecoceutics.com\/ca\/respostes-de-salut\/salut\/funcionament-pcr\/#que-es-la-rt-pcr\" >Qu\u00e8 \u00e9s la RT-PCR?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-4\" href=\"https:\/\/www.ecoceutics.com\/ca\/respostes-de-salut\/salut\/funcionament-pcr\/#que-es-la-rrt-pcr\" >Qu\u00e8 \u00e9s la&nbsp;rRT-PCR?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-5\" href=\"https:\/\/www.ecoceutics.com\/ca\/respostes-de-salut\/salut\/funcionament-pcr\/#quines-limitacions-presenta-la-pcr\" >Quines limitacions presenta la PCR?<\/a><\/li><\/ul><\/nav><\/div>\n<h2><span class=\"ez-toc-section\" id=\"que-es-la-reaccio-en-cadena-de-la-polimerasa\"><\/span>Qu\u00e8 \u00e9s la reacci\u00f3 en cadena de la polimerasa?<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>\u00c9s la repetici\u00f3 successiva d\u2019una reacci\u00f3 qu\u00edmica en la qual s\u2019aprofita un enzim anomenat polimerasa per a generar m\u00e9s ADN a partir d\u2019una mostra de material gen\u00e8tic original. Per a entendre-la b\u00e9 hem de con\u00e8ixer uns detalls de l\u2019ADN pr\u00e8viament, aix\u00ed que comencem pel principi.<\/p>\n<p>L\u2019ADN \u00e9s una macromol\u00e8cula, \u00e9s a dir, una mol\u00e8cula gegant, composta d\u2019unitats m\u00e9s petites anomenades \u00abnucle\u00f2tids\u00bb. Els nucle\u00f2tids s\u2019uneixen qu\u00edmicament per a formar una cadena, i dues cadenes complement\u00e0ries s\u00f3n capaces d\u2019unir-se i tor\u00e7ar-se sobre si mateixes per a formar una doble h\u00e8lix. Entre altres propietats de l\u2019ADN, comptem amb:<\/p>\n<ul>\n<li>La doble h\u00e8lix s\u2019obre en escalfar-se, i s\u2019obtenen dues cadenes simples de nucle\u00f2tids.<\/li>\n<li>Cadascuna d\u2019aquestes cadenes pot servir de motlle per a la cadena companya que forma la doble h\u00e8lix, perqu\u00e8 s\u00f3n cadenes perfectament complement\u00e0ries. Aquesta propietat permet que l\u2019ADN sigui \u00abautocopiant\u00bb amb els enzims de s\u00edntesis adequades, sent una de la m\u00e9s important l\u2019enzim anomenat ADN polimerasa.<\/li>\n<li>L\u2019ADN polimerasa pot copiar una cadena utilitzant nucle\u00f2tids individuals solts com si fos un paleta que fa una paret a partir de maons, per\u00f2 necessita almenys tenir un trosset de cadena complement\u00e0ria disponible per a comen\u00e7ar a sintetitzar una nova cadena. A aquest trosset l\u2019anomenem primer.<\/li>\n<\/ul>\n<p>La reacci\u00f3 en cadena de la polimerasa aprofita aquestes propietats per a obtenir una bona quantitat de material gen\u00e8tic a partir de poques (o fins i tot d\u2019una) dobles h\u00e8lixs.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"com-funciona-la-reaccio-en-cadena-de-la-polimerasa\"><\/span>Com funciona la reacci\u00f3 en cadena de la polimerasa?<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>Suposem que partim d\u2019una sola cadena doble d\u2019ADN trobada en una mostra biol\u00f2gica (sang, pell, secrecions nasals, etc.). A aquesta mostra podem afegir-li polimerasa, primers i nucle\u00f2tids, i dur a terme els seg\u00fcents passos amb un equip automatitzat:<\/p>\n<ul>\n<li>Escalfament de la mostra a una temperatura tal que s\u2019obri la doble h\u00e8lix, per\u00f2 no es destrueixi (90-95 \u00b0C).<\/li>\n<li>Refredament lleuger del medi de reacci\u00f3, perqu\u00e8 s\u2019uneixin les cadenes simples amb els primers disponibles en el medi.<\/li>\n<li>S\u00edntesi d\u2019una nova cadena complement\u00e0ria a les anteriors. Aix\u00f2 es realitza a trav\u00e9s de polimerases capaces de tolerar l\u2019escalfament i de treballar a 60-75 \u00b0C, com la polimerasa provinent dels microorganismes&nbsp;Thermus&nbsp;aquaticus&nbsp;(TaqDNA&nbsp;polimerasa, la primera a usar-se) o&nbsp;Pyrococcus&nbsp;furiosus&nbsp;(PfuDNA&nbsp;polimerasa, una miqueta millor copiant que la primera).<\/li>\n<\/ul>\n<p>En finalitzar aquests passos, en comptes d\u2019una doble cadena d\u2019ADN tenim dues cadenes d\u2019ADN, perqu\u00e8 cada cadena es va usar com a motlle per a una nova doble h\u00e8lix. Si afegim a aquest cicle altre igual, en comptes de quatre cadenes tindrem vuit. I si encadenem molts cicles (\u00abreacci\u00f3 en cadena\u00bb) obtindrem una quantitat important d\u2019ADN amb el qual podrem diagnosticar una infecci\u00f3, fer una prova de paternitat, trobar al culpable d\u2019un assassinat, etc. I si controlem acuradament les condicions de reacci\u00f3, podem tenir una idea de quant material gen\u00e8tic ten\u00edem en comen\u00e7ar: en el nostre exemple, si necessitem partim de 50 dobles h\u00e8lixs d\u2019ADN necessitarem un cicle per a arribar a 100 cadenes, per\u00f2 si partim d\u2019una sola, necessitarem diversos cicles (una h\u00e8lix resulta en dues, dues resulten en quatre, quatre resulten en vuit\u2026.i aix\u00ed successivament).<\/p>\n<h2><span class=\"ez-toc-section\" id=\"que-es-la-rt-pcr\"><\/span>Qu\u00e8 \u00e9s la RT-PCR?<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>La RT-PCR \u00e9s una modificaci\u00f3 de la t\u00e8cnica de la PCR necess\u00e0ria per a treballar amb material gen\u00e8tic de tipus ARN, com el que es troba en el virus&nbsp;Sars-CoV-2&nbsp;o el VIH. La polimerasa nom\u00e9s pot treballar amb ADN, per la qual cosa necessitem un enzim que sintetitzi una cadena d\u2019ADN complement\u00e0ria a la cadena d\u2019ARN original; aquesta funci\u00f3 la compleix l\u2019enzim anomenat transcriptasa&nbsp;reversa. El primer pas de la RT-PCR \u00e9s la s\u00edntesi de l\u2019ADN complementari a l\u2019ARN original i la destrucci\u00f3 de l\u2019ARN, per a despr\u00e9s fer els mateixos cicles de la PCR explicats a dalt.<\/p>\n<p>El nom RT-PCR ve de l\u2019angl\u00e8s&nbsp;reverse&nbsp;transcription&nbsp;\u2013&nbsp;polymerase&nbsp;chain&nbsp;reaction.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"que-es-la-rrt-pcr\"><\/span>Qu\u00e8 \u00e9s la&nbsp;rRT-PCR?<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>La&nbsp;r&nbsp;min\u00fascula \u00e9s el s\u00edmbol de \u00abreal time\u00bb, \u00e9s a dir, \u00aben temps real\u00bb. \u00c9s una modificaci\u00f3 de la RT-PCR que permet fer un seguiment detallat dels cicles de reacci\u00f3 i \u00e9s molt \u00fatil per a quantificar el material gen\u00e8tic de la mostra.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"quines-limitacions-presenta-la-pcr\"><\/span>Quines limitacions presenta la PCR?<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p>La PCR i les t\u00e8cniques relacionades s\u00f3n molt fiables, per\u00f2 presenten alguns inconvenients:<\/p>\n<ul>\n<li>Requereixen temps. Es requereixen diverses hores per a obtenir el resultat d\u2019un pacient, limitant la capacitat d\u2019un laboratori per a lliurar resultats.<\/li>\n<li>Alta sensibilitat a la contaminaci\u00f3. Els laboratoris solen treballar en \u00e0rees separades per a la&nbsp;pre-amplificaci\u00f3&nbsp;i per a la&nbsp;post-amplificaci\u00f3, perqu\u00e8 la contaminaci\u00f3 amb poquetes cadenes d\u2019ADN pot donar un fals positiu. La PCR sol&nbsp;fer-se amb&nbsp;un control negatiu (reacci\u00f3 sense material gen\u00e8tic present) que, si d\u00f3na positiu, delata una possible contaminaci\u00f3 del sistema de reacci\u00f3.<\/li>\n<li>S\u00f3n costoses. L\u2019aparell que fa aquestes proves no \u00e9s barat, els materials per a dur a terme la prova tampoc s\u00f3n econ\u00f2mics i es requereix personal qualificat per a operar-ho i interpretar el resultat correctament.<\/li>\n<\/ul>\n<p>\u00c9s per aix\u00f2 que, en moments cr\u00edtics com pot ser la pand\u00e8mia de covid-19, s\u2019han buscat opcions m\u00e9s r\u00e0pides i econ\u00f2miques a la PCR com els tests d\u2019ant\u00edgens, que estan llestos en 15 minuts i no requereixen tant d\u2019equip o instal\u00b7lacions.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>La reacci\u00f3 en cadena de la polimerasa (la famosa PCR) \u00e9s una t\u00e8cnica que s\u2019utilitza per a \u00abamplificar\u00bb petites quantitats d\u2019ADN. La sigla PCR ve del nom en angl\u00e8s de la t\u00e8cnica:&nbsp;Polymerase&nbsp;Chain&nbsp;Reaction. El material gen\u00e8tic en petita quantitat resulta invisible per als m\u00e8todes d\u2019an\u00e0lisis, fins i tot els m\u00e9s moderns, per la qual cosa \u00e9s [&hellip;]<\/p>\n","protected":false},"author":4,"featured_media":17978,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"inline_featured_image":false,"_mbp_gutenberg_autopost":false,"footnotes":""},"categories":[4,1],"tags":[],"class_list":["post-16525","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-respostes-de-salut","category-salut"],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.4 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>En qu\u00e8 consisteix una PCR? | farm\u00e0cies ecoceutics<\/title>\n<meta name=\"description\" content=\"Saps qu\u00e8 signifiquen les sigles PCR? \u00c9s la reacci\u00f3 en cadena de la polimerasa, un proc\u00e9s per amplificar l&#039;ADN. 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