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Performance of NIST SRM® 2917 with 13 Recreational Water Quality Monitoring qPCR Assays (ASM Microbe 2022)
Citation:
Willis, J., Mano Sivaganesan, Rich Haugland, J. Kralj, S. Servetas, M. Hunter, AND S. Jackson. Performance of NIST SRM® 2917 with 13 Recreational Water Quality Monitoring qPCR Assays (ASM Microbe 2022). ASM Microbe 2022, Washington, DC, June 09 - 13, 2022.
Impact/Purpose:
National implementation of real-time quantitative PCR recreational water quality monitoring tools requires the development of and access to a high-quality standard control material. This manuscript reports a single laboratory qPCR performance assessment of the National Institute of Science and Technology Standard Reference Material 2917 (NIST SRM® 2917), a linearized plasmid DNA construct that functions with 13 recreational water quality qPCR assays. Findings demonstrated that NIST SRM® 2917 functions with all qPCR methods and suggests that the future use of this control material by scientists and water quality managers should help reduce variability in concentration estimates and make results more consistent between laboratories. This effort addresses an EPA Office of Water high research priority described in the ORD Research Action Plan (SSWR 3.1.1).
Description:
Fecal pollution remains a significant challenge for recreational water quality management worldwide. In response, there is a growing interest in the use of real-time quantitative PCR (qPCR) methods to achieve same-day notification of recreational water quality and associated public health risk as well as to characterize fecal pollution sources for targeted mitigation. However, successful widespread implementation of these technologies requires the development of and access to a high-quality standard control material. Here, we report a single laboratory qPCR performance assessment of the National Institute of Science and Technology Standard Reference Material 2917 (NIST SRM® 2917), a linearized plasmid DNA construct that functions with 13 recreational water quality qPCR assays. Performance experiments indicate the generation of standard curves with amplification efficiencies ranging from 0.95 ± 0.006 to 0.99 ± 0.008 and coefficient of determination values (R2) ≥ 0.980. Regardless of qPCR assay, variability in repeated measurements at each dilution level were very low (cycle threshold standard deviations ≤ 0.657) and exhibited a heteroscedastic trend characteristic of qPCR standard curves. The influence of a yeast carrier tRNA added to the standard control material buffer was also investigated. Findings demonstrated that NIST SRM® 2917 functions with all qPCR methods and suggests that the future use of this control material by scientists and water quality managers should help reduce variability in concentration estimates and make results more consistent between laboratories.