PCR-Based Diagnosis of Helicobacter pylori Infection and Real-Time Determination of Clarithromycin Resistance Directly from Human Gastric Biopsy Samples
| Autor: | Stephanie A. Chisholm, E. Louise Teare, Robert Owen, Seth Saverymuttu |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
DNA
Bacterial Microbiology (medical) Biopsy Drug resistance Polymerase Chain Reaction Helicobacter Infections law.invention Bacterial Proteins law 23S ribosomal RNA Clarithromycin RNA Ribosomal 16S medicine Humans Helicobacter Polymerase chain reaction Helicobacter pylori biology medicine.diagnostic_test Hybridization probe Stomach Bacteriology Drug Resistance Microbial bacterial infections and mycoses biology.organism_classification Virology Anti-Bacterial Agents RNA Ribosomal 23S medicine.drug |
| Zdroj: | Journal of Clinical Microbiology. 39:1217-1220 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.39.4.1217-1220.2001 |
| Popis: | A novel PCR detection assay that amplifies the Helicobacter pylori -specific vacuolating cytotoxin gene ( vacA ) and thus enables rapid diagnosis of infection is described. Additionally, a real-time probe hybridization melting point analysis assay to detect all three mutations in the 23S rRNA gene associated with clarithromycin resistance was applied directly to antral gastric biopsy samples. Comparison with culture and an alternative PCR assay targeting the 16S rrn gene showed that the vacA assay was sensitive and specific when tested on biopsy samples from 121 patients. Clarithromycin susceptibilities could be determined in the majority (92.3%) of culture-positive gastric biopsy samples analyzed, four of which generated melting peaks indicative of clarithromycin resistance by either an A→G or A→C mutation. The presence of the mutations correlated with the clarithromycin disk diffusion sensitivities of matched cultures. This PCR-based system was simple to perform and could be completed in 3 to 4 h, thereby overcoming the delays associated with conventional culture methods for H. pylori identification and susceptibility testing. |
| Databáze: | OpenAIRE |
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