| Titre : | Validation d’une PCR en temps réel qualitative permettant la détection de Campylobacter species sur des prélèvements de selles humaines |
| Auteurs : | Yasmina El Kanchidi, Auteur ; Benoit Prevost, Promoteur |
| Type de document : | Travail de fin d'études |
| Editeur : | Woluwe-Saint-Lambert : Haute École Léonard de Vinci, 2024 |
| Index. décimale : | TFE - Biologie médicale |
| Mots-clés: | Campylobacter ; Réaction de polymérisation en chaine en temps réel ; ARN ribosomique 16S ; Recherche qualitative ; Etude de validation de principe |
| Résumé : | This research study presents the validation results of a real-time qualitative PCR method for detecting Campylobacter species in human stool samples. Globally, an estimated 550 million people, including 220 million children under the age of 5, experience Campylobacter species gastroenteritis annually. These bacteria are challenging to culture due to their slow growth rate and varying incubation conditions depending on the species. Hence, PCR is employed in the laboratory to expedite the process and deliver a prompt qualitative analysis result to patients. The method involves two primary stages. The first stage is the extraction process utilizing the EZ2 equipment, which purifies bacterial DNA through elution with magnetic silica beads. The second stage is an in-house duplex PCR performed with the Quantstudio instrument, targeting two sequences: the Campylobacter species 16S rRNA and an internal control added to each sample. The goal of this method is to ascertain the presence or absence of the bacterium in a patient sample, without quantifying it. The focus is solely on correlating the symptomatology of gastroenteritis patients with the bacterial cause. The results for various validation criteria were conclusive, except for one temperature point in terms of reproducibility. Consequently, the method still requires further review and refinement by the research laboratory. |
| Disponible en ligne : | Oui |
| Lieu du stage : | LHUB-ULB site Anderlecht, service de microbiologie, secteur MICN, CNR Campylobacter |
| Département du TFE : | Biologie médicale |
Documents numériques (1)
 TFE Adobe Acrobat PDF |
