Clinical personnel and health officials must rapidly identify potential pathogens in order to respond to suspected disease outbreaks to prevent public disturbances. Next-generation sequencing has the potential to complement conventional diagnostics in many instances, and whole genome sequencing is ideal for use as a typing tool to identify the causative agent behind diseases.
One recent example of NGS being used in a clinical setting is illustrated in the publication Rapid Metagenomic Diagnostics for Suspected Outbreak of Severe Pneumonia, a letter to Emerging Infectious Diseases, in which researchers describe the methodology their group used to identify the pathogen responsible for acute respiratory distress syndrome in their patients, using a NEXTflex™ Kit for library construction. This sequencing methodology was used to identify the etiologic agent of a suspected pneumonia outbreak among German police officers.
With a robust two-hour library prep protocol and up to 384 single-index barcodes, which facilitate high level multiplexing while reducing concerns about run-to-run sample carryover and sample crosstalk, the NEXTflex™ Rapid DNA-Seq Kit is ideal for rapid identification of disease-causing pathogens.
To demonstrate the efficacy of next generation sequencing for identification of pathogens, we present below Bioanalyzer traces of libraries constructed from varying inputs of DNA isolated from E. coli, Campylobacter and Salmonella bacteria.
Figure 1. Bioanalyzer trace of a library constructed using the NEXTflex Rapid DNA-Seq Kit from 100 ng of E. coli bacteria in water.
Figure 2. Bioanalyzer trace of a library constructed using the NEXTflex Rapid DNA-Seq Kit from 1 µg of Campylobacter bacteria in water.
Figure 3. Bioanalyzer trace of a library constructed using the NEXTflex Rapid DNA-Seq Kit from 100 ng Salmonella bacteria in water.
Fischer, N., et al. (2014) Rapid metagenomic diagnostics for suspected outbreak of severe pneumonia [letter]. Emerg Infect Dis. doi: 10.3201/eid2006.131526.