| Popis: |
Objective: Timely diagnosis is crucial for successful treatment of infections. Contemporarydiagnostics of clinically significant infections relies on a variety of laboratory-based tests.However, the routine microbiological methods, per se are not always confirmatory for theaccurate diagnosis in some cases with a significant time delay. Rapid detection of causativeagents using nanopore-based metagenomic sequencing is reported in a case of suspectedtuberculosis not confirmed using conventional microbiology diagnostics, and in a case ofsuspected brucellosis with delayed confirmation by conventional microbiology.Materials and Methods: This report includes two cases. Patient-1 was clinically diagnosedwith peritoneal tuberculosis while Patient-2 had relapsing brucellosis. Cultures and PCRwere negative for the peritoneal fluid of Patient-1. Serological determinants for brucellosiswere under the thresholds for Patient-2. The blood culture was positive for Brucella spp;however, the results were available only after seven days. Peritoneal fluid and blood samplewere obtained from Patient-1 and Patient-2, respectively. Total DNA was extracted usingcommercial kits. Shotgun metagenomic sequencing was performed using Oxford NanoporeTechnologies. Read-event data were base-called by the software Metrichor using workflowv2.39.3.Results: For Patient-1, the first Mycobacterium tuberculosis reads were confidently detected in20 minutes, for Patient-2 the first Brucella melitensis read was detected at 30-minutes-timepoint. Overall, the pathogens were detected within 6 hours. We demonstrated a real-timenanopore sequencing technology for the rapid detection of infectious agents in two crucialcases where traditional microbiological methods failed to reveal the pathogens.Conclusion: In this report, we present that real-time sequencing can not only detectconventionally unrevealed pathogens, but it can achieve this in a tight time-frame.Therefore, it could be said that the nanopore sequencing has the promise to enhance ourability to diagnose infectious diseases. |
