{"id":24517,"date":"2021-03-19T15:43:00","date_gmt":"2021-03-19T14:43:00","guid":{"rendered":"https:\/\/www.chu-poitiers.fr\/?p=24517"},"modified":"2021-05-18T10:43:36","modified_gmt":"2021-05-18T08:43:36","slug":"covid-19-identifier-les-variants","status":"publish","type":"post","link":"https:\/\/www.chu-poitiers.fr\/en\/covid-19-identifier-les-variants\/","title":{"rendered":"Covid-19 : identifier les variants"},"content":{"rendered":"\n<h3>Avant de proc\u00e9der au s\u00e9quen\u00e7age, il faut d\u2019abord attester la pr\u00e9sence du virus. Pouvez-vous nous dire comment cela se passe&nbsp;?<\/h3>\n<p>Lorsque les \u00e9chantillons arrivent \u00e0 la plateforme covid, ils sont d\u2019abord soumis \u00e0 un test RT-PCR de d\u00e9pistage qui confirme ou non l\u2019infection au SARS-CoV2. Pour cela, une fois inactiv\u00e9s et enregistr\u00e9s, les \u00e9chantillons sont plac\u00e9s dans un automate qui les d\u00e9pose sur une plaque. Un autre automate extrait l\u2019ARN, l\u2019information g\u00e9n\u00e9tique du virus. La plaque est ensuite plac\u00e9e dans une machine qui permet d\u2019amplifier trois g\u00e8nes cibles du virus au travers de cycles d\u2019augmentation et de baisse de temp\u00e9ratures de 40\u00b0C \u00e0 90\u00b0C. Trois g\u00e8nes sont cibl\u00e9s parmi lesquels un fragment de la prot\u00e9ine S qui permet au virus de p\u00e9n\u00e9trer dans les cellules. Un logiciel permet de contr\u00f4ler en temps r\u00e9el \u00e0 chaque cycle les courbes d\u2019amplification des trois g\u00e8nes figur\u00e9es par des signaux fluorescents. L\u2019apparition de ces signaux indique la pr\u00e9sence du virus.<\/p>\n<h3>A cette \u00e9tape, pouvez-vous d\u00e9terminer le type de variant&nbsp;?<\/h3>\n<p>Apr\u00e8s cette premi\u00e8re \u00e9tape d\u2019analyse RT-PCR, nous pouvons dire, d\u2019une part, si l\u2019\u00e9chantillon est infect\u00e9 ou non au SARS-CoV2 et indiquer les \u00e9chantillons susceptibles d\u2019\u00eatre des variants britanniques. La pr\u00e9sence des trois g\u00e8nes cibl\u00e9s confirme qu\u2019il s\u2019agit du SARS-CoV2. Par contre, l\u2019absence de d\u00e9tection de la prot\u00e9ine S correspond \u00e0 un profil suspect du variant britannique puisque le fragment que nous ciblons contient une premi\u00e8re mutation, la d\u00e9l\u00e9tion 69-70, caract\u00e9ristique de ce variant. A ce stade, nous ne pouvons cependant pas affirmer qu\u2019il s\u2019agisse bien du variant britannique. De plus, il est impossible de pr\u00e9dire de potentiels variants sud-africains ou br\u00e9siliens qui sont par ailleurs surveill\u00e9s. C\u2019est pourquoi nous allons entreprendre un autre test RT-PCR dit de criblage sur tous les pr\u00e9l\u00e8vements positifs au SARS-CoV2.<\/p>\n<h3>Que sont ces tests RT-PCR de criblage ?<\/h3>\n<p>Ce type de test d\u00e9tecte une autre mutation de la prot\u00e9ine S connue du virus, la substitution N501Y que l\u2019on retrouve chez les variants britannique, br\u00e9silien et sud-africain. Ce test s\u2019effectue sur les m\u00eames automates que le d\u00e9pistage mais utilise d\u2019autres r\u00e9actifs pour l\u2019amplification de l\u2019ARN. Les tests RT-PCR de criblage est une alternative, plus rapide et \u00e0 plus haut d\u00e9bit, \u00e0 la m\u00e9thode classique de s\u00e9quen\u00e7age. En effet, le s\u00e9quen\u00e7age de certaines portions du g\u00e9nome viral ne peut \u00eatre r\u00e9alis\u00e9 que pour une vingtaine d\u2019\u00e9chantillons par semaine alors que le criblage permet de tester jusqu\u2019\u00e0 200 pr\u00e9l\u00e8vements par jour. N\u00e9anmoins, le s\u00e9quen\u00e7age est une technique susceptible d\u2019identifier un plus grand nombre de mutations que le criblage. On le r\u00e9serve donc \u00e0 une analyse plus fine des pr\u00e9l\u00e8vements suspects de contenir un variant sud-africain ou br\u00e9silien afin d\u2019obtenir une identification d\u00e9finitive.<\/p>\n<h3><strong>Quelles m\u00e9thodes de s\u00e9quen\u00e7age utilisez-vous ?<\/strong><\/h3>\n<p>A ce jour, nous utilisons la m\u00e9thode dite Sanger utilis\u00e9e en virologie pour le g\u00e9notypage des souches de VIH \u00e0 la recherche de mutations de r\u00e9sistance aux traitements antiviraux. Cette technique recherche \u00e0 des endroits pr\u00e9cis du g\u00e9nome du virus les mutations caract\u00e9ristiques des variants. L\u2019ARN des \u00e9chantillons est extrait et deux r\u00e9gions couvrant ces mutations caract\u00e9ristiques sont amplifi\u00e9es puis plac\u00e9es dans un s\u00e9quenceur qui les lit. Nous obtenons alors, pour chaque \u00e9chantillon, les s\u00e9quences nucl\u00e9otidiques (enchainement des nucl\u00e9otides ATCG constitutif de l\u2019ADN) des deux g\u00e8nes cibl\u00e9s que nous comparons \u00e0 la s\u00e9quence de r\u00e9f\u00e9rence du virus, tel qu\u2019il est apparu en Chine au d\u00e9but de la pand\u00e9mie. Tous les chercheurs utilisent cette m\u00eame s\u00e9quence de r\u00e9f\u00e9rence. La comparaison \u00e0 cette r\u00e9f\u00e9rence nous permet alors d\u2019identifier des mutations ou diff\u00e9rences dans les s\u00e9quences analys\u00e9es et de d\u00e9terminer quel variant est pr\u00e9sent dans un \u00e9chantillon. Nous avons ainsi d\u00e9tect\u00e9 les premiers cas de variant sud-africain en Vienne d\u00e9but f\u00e9vrier.<\/p>\n<p>Mais cette m\u00e9thode de s\u00e9quen\u00e7age a ses limites puisqu\u2019elle ne permet de s\u00e9quencer qu\u2019une toute petite partie du g\u00e9nome du virus. Or, il existe des \u00e9quipements de s\u00e9quen\u00e7age \u00e0 haut-d\u00e9bit, d\u00e9j\u00e0 utilis\u00e9s en canc\u00e9rologie ou en g\u00e9n\u00e9tique, qui permettent de s\u00e9quencer le g\u00e9nome entier du SARS-CoV2. Ces \u00e9quipements innovants g\u00e9n\u00e8rent une quantit\u00e9 astronomique de s\u00e9quences qu\u2019il faut analyser par le crible d\u2019outils bio-informatiques pour obtenir les donn\u00e9es essentielles pour nos recherches. Etudier le g\u00e9nome complet nous permettrait de r\u00e9aliser une meilleure surveillance \u00e9pid\u00e9miologique du virus et notamment l\u2019identification de nouveaux variants susceptibles d\u2019appara\u00eetre \u00e0 la suite de la g\u00e9n\u00e9ralisation de la vaccination. Nous allons mettre en place ce s\u00e9quen\u00e7age \u00e0 haut-d\u00e9bit mi-f\u00e9vrier.<\/p>\n[1] D\u00e9tentrice d\u2019un doctorat en biologie de l\u2019Institut de Recherche sur la Biologie de l\u2019Insecte, Caroline Michaud a des comp\u00e9tences en biologie mol\u00e9culaire qui fait appel aux m\u00eames outils de s\u00e9quen\u00e7ages.<\/p>\n\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-large is-resized\"><a href=\"https:\/\/www.chu-poitiers.fr\/wp-content\/uploads\/2021\/05\/Dr-Michaud-1-1024x1019-1.jpg\"><img decoding=\"async\" src=\"https:\/\/www.chu-poitiers.fr\/wp-content\/uploads\/2021\/05\/Dr-Michaud-1-1024x1019-1.jpg\" alt=\"\" class=\"wp-image-24532\" width=\"512\" height=\"510\"\/><\/a><\/figure><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Une course contre la montre est lanc\u00e9e afin de freiner la propagation des variants du SARS-CoV2 consid\u00e9r\u00e9s plus contagieux. Les pouvoirs publics\u2026","protected":false},"author":12,"featured_media":24541,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[96],"tags":[],"class_list":["post-24517","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-non-classifiee"],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v28.6 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Covid-19 : identifier les variants - Site du CHU de Poitiers<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.chu-poitiers.fr\/covid-19-identifier-les-variants\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Covid-19 : identifier les variants - Site du CHU de Poitiers\" \/>\n<meta property=\"og:description\" content=\"Une course contre la montre est lanc\u00e9e afin de freiner la propagation des variants du SARS-CoV2 consid\u00e9r\u00e9s plus contagieux. Les pouvoirs publics ont mobilis\u00e9 les laboratoires hospitaliers dans la traque aux variants. Depuis janvier, le laboratoire de virologie du CHU de Poitiers s\u2019adapte et utilise des techniques d\u2019identification pour un suivi \u00e9pid\u00e9miologique plus pr\u00e9cis. 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Les pouvoirs publics ont mobilis\u00e9 les laboratoires hospitaliers dans la traque aux variants. Depuis janvier, le laboratoire de virologie du CHU de Poitiers s\u2019adapte et utilise des techniques d\u2019identification pour un suivi \u00e9pid\u00e9miologique plus pr\u00e9cis. 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