Constantin Hays1, Thibault Delerue1, Dominique Lamarque2, Christophe Burucoa3, Ghislaine Collobert1, Annick Billöet1, Nicolas Kalach4, Josette Raymond1. 1. Bacteriology, Cochin Hospital, University of Paris-Descartes, Institut Pasteur, Paris, France. 2. Gastroentérologie, Hôpital Ambroise Paré, Université Versailles- Saint Quentin, Boulogne, France. 3. Laboratoire de bactériologie, Hygiène, EA 4331 LITEC, CHU de Poitiers, Université de Poitiers, Poitiers, France. 4. Clinique pédiatrique Saint Antoine, Hôpital Saint Vincent de Paul, Groupement des Hôpitaux de l'Institut Catholique de Lille (GHICL), Lille, France.
Abstract
BACKGROUND: Adapted treatments for Helicobacter pylori infection, guided by determining antimicrobial resistance, are associated with high eradication rates. We evaluated the performance of the Amplidiag® H. pylori + ClariR PCR assay (Amplidiag® ) for detecting H. pylori and its clarithromycin resistance from gastric biopsies taken during endoscopy in comparison to culture and our "in-house" PCR. MATERIALS AND METHODS: A total of 127 gastric biopsies were analyzed (98 adults; 29 children). Culture, PCR Amplidiag® , and in-house PCR were performed in parallel. The in-house PCR combined amplification and sequencing of a 267-bp fragment of the H. pylori 23S rRNA gene. Discrepancies were controlled by amplification of glmM gene. RESULTS: For detection of H. pylori, Amplidiag® and the in-house PCR were concordant in 118 of 127 of cases: 66 negative and 52 positive. Discrepancies were observed in nine cases, all with low bacterial load: Amplidiag® did not detect seven biopsies positive on in-house PCR but detected two positive biopsies that were negative on in-house PCR. Among the 19 of 52 (36%) H. pylori cases resistant to clarithromycin, only four biopsies with mixed populations exhibited discordant results between the two PCR methods. The A2142T mutation was not detected by Amplidiag® . With the in-house PCR and amplified glmM gene as the reference method, the sensitivity and specificity of Amplidiag® was 88.5% (95% confidence interval 83-94.1) and 100%. CONCLUSION: This study demonstrated the high sensitivity of the PCR-based Amplidiag® H. pylori test, especially with low H. pylori load, and the probability of its clarithromycin resistance analysis. For clinical use, a well-designed trial with a large scale of samples may still be needed.
BACKGROUND: Adapted treatments for Helicobacter pylori infection, guided by determining antimicrobial resistance, are associated with high eradication rates. We evaluated the performance of the Amplidiag® H. pylori + ClariR PCR assay (Amplidiag® ) for detecting H. pylori and its clarithromycin resistance from gastric biopsies taken during endoscopy in comparison to culture and our "in-house" PCR. MATERIALS AND METHODS: A total of 127 gastric biopsies were analyzed (98 adults; 29 children). Culture, PCR Amplidiag® , and in-house PCR were performed in parallel. The in-house PCR combined amplification and sequencing of a 267-bp fragment of the H. pylori 23S rRNA gene. Discrepancies were controlled by amplification of glmM gene. RESULTS: For detection of H. pylori, Amplidiag® and the in-house PCR were concordant in 118 of 127 of cases: 66 negative and 52 positive. Discrepancies were observed in nine cases, all with low bacterial load: Amplidiag® did not detect seven biopsies positive on in-house PCR but detected two positive biopsies that were negative on in-house PCR. Among the 19 of 52 (36%) H. pylori cases resistant to clarithromycin, only four biopsies with mixed populations exhibited discordant results between the two PCR methods. The A2142T mutation was not detected by Amplidiag® . With the in-house PCR and amplified glmM gene as the reference method, the sensitivity and specificity of Amplidiag® was 88.5% (95% confidence interval 83-94.1) and 100%. CONCLUSION: This study demonstrated the high sensitivity of the PCR-based Amplidiag® H. pylori test, especially with low H. pylori load, and the probability of its clarithromycin resistance analysis. For clinical use, a well-designed trial with a large scale of samples may still be needed.
Authors: Rebecca Marrero Rolon; Scott A Cunningham; Jayawant N Mandrekar; Erin T Polo; Robin Patel Journal: J Clin Microbiol Date: 2021-04-20 Impact factor: 5.948