| Titre : | Evaluation d'un kit PCR en temps réel pour le diagnostic de l'infection à helicobacter pylori et la résistance à la claritromycine |
| Auteurs : | Ouzeifatou Ouedraogo, Auteur ; Véronique Yvette Miendje Deyi, 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: | Helicobacter infection / microbiologie ; Helicobacter infection / thérapie ; Helicobacter infection/ transmission ; Helicobacter infection/ pathologie ; Clarithromycine /pharmacologie ; Helicobacter pylori ; Helicobacter infection /thérapie médicamenteuse ; Clarithromycine ; Diagnostic ; Traitement |
| Résumé : |
Background: Helicobacter pylori is a microaerophilic Gram-positive bacillus that colonizes the gastric mucosa of around half of the world's population. Early diagnosis and appropriate treatment can prevent the onset of pathologies caused by H. pylori such as peptic ulcers and gastric cancer. Culture is the reference method, but it is fastidious and depends on rigorous pre-analytical conditions for H.pylori isolation. This work was carried out in this context, in order to implement the use of molecular methods, less dependent on pre-analytical conditions.
Aim: The aim of this study is to validate a PCR kit for the diagnosis of H.pylori infection and the detection of mutations (A2142C, A2142G and A2143G) involved in clarithromycin resistance, a key antibiotic in treatment. Material and methods: Gastric biopsies will be tested in parallel by molecular method (real-time PCR: allplexTM H.pylori & ClariR Assay) and phenotypic method (culture using selective media and determination of clarithromycin susceptibility by diffusion disk method or E-test). Discordance cases will be analyzed, compared with histology results, and retested if necessary. Concordance rates will be calculated for both diagnosis and resistance detection. Performance tests (detection limit, repeatability, reproducibility and quality control) will also be carried out. Results and discussion: A total of 58 gastric biopsies were processed in parallel. We observed a final concordance of 71.41% for the diagnosis of H. pylori infection and 97.30% for the determination of clarithromycin susceptibility. Analysis of the origin of the 16 cases of discordance for diagnosis showed that culture negativity could be attributed to a break in the cold chain. Nevertheless, 14 (87.5%) of these cases of discordance also had a negative histology result, which is rather in favor of culture efficiency. For resistance detection, one case of discordance (absence of mutations and resistant with disc/E-test) was observed. This remains to be clarified by further analysis. The performance tests carried out were acceptable for PCR. Moreover, the detection limit for PCR was very low, compared to culture. Conclusion and perspectives: At the end of this work, the results obtained with the Real time PCR method (allplexTM H.pylori & ClariR Assay) are conclusive. After further analysis, routine implementation of PCR would enable faster diagnostic results, but also more rapid detection of sensitivity to clarithromycin. PCR would probably have to be coupled with culture to obtain susceptibility results for other antibiotics used for H.pylori eradication. To be discus by Microbiologists taking in account the entire data. |
| Disponible en ligne : | Oui |
| Lieu du stage : | LHUB-ULB Porte de Hal site (Rue haute 322 ,1000 Bruxelles |
| Département du TFE : | Biologie médicale |
Documents numériques (1)
 TFE Adobe Acrobat PDF |
