Comparison of next-generation droplet digital PCR with quantitative PCR for enumeration of Naegleria fowleri in environmental water and clinical samples

Abstract

This study explored the application of ddPCR and qPCR methods in identifying Naegleria fowleri from both clinical and environmental water samples. Strong agreement between ddPCR and qPCR methods over clinical DNA samples was observed. Naegleria fowleri was present in surface water samples from Lake Pontchartrain during our study period. The ability of N. fowleri to survive in brackish water is therefore a potential risk factor for people who engage in water-related recreational activities. The ddPCR performance demonstrated in this study on clinical and environmental samples lead to greater confidence of ddPCR technology on field application.

Quantitative Microbial Risk Assessment and Molecular Biology: Paths to Integration

“…Some of this may relate to the issue of viability discussed in the next section. For example, Xue et al 33 found close correspondence between qPCR and dPCR measurements of Naegleria fowleri in spiked water. In contrast, Ricchi et al 34 analyzed water samples spiked with three strains each of three bacteria using plate count, qPCR, and dPCR (on two instruments) as shown in Figure 2.…”

“…Some of this may relate to the issue of viability discussed in the next section. For example, Xue et al found close correspondence between qPCR and dPCR measurements of Naegleria fowleri in spiked water. In contrast, Ricchi et al .…”

Integrating Virus Monitoring Strategies for Safe Non-Potable Water Reuse

“…High-throughput qPCR using microfluidic technology is demonstrated as a direct multi-pathogen detection approach for environmental water samples. This technology makes use of microfluidic chips, which allow for high-throughput measurement of large sample quantities for a variety of enteric viruses and other pathogens [53,54].…”

“…Droplet digital PCR (ddPCR) is also shown to have improved performance in the presence of inhibitory compounds as compared to qPCR [54,55]. ddPCR performs better because it is an end-point positive/negative detection combined with Poisson statistics for quantification, so it has higher accuracy and precision against PCR inhibition.…”

Detection of Kudoa thyrsites (Myxozoa) eDNA by real‐time and digital PCR from high seawater volumes

“…dPCR offers lower impacts from sample inhibitors, independence from calibration curves, high levels of precision (Pavsic et al, 2017) and may improve sensitivity (e.g. Xue et al, 2018). Finally, we compared both platforms using samples collected from an active farm with a history of high K. thyrsites impact.…”