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Interlaboratory performance and quantitative PCR data acceptance metrics for NIST SRM® 2917
Citation:
Sivaganesan, M., J. Willis, M. Karim, A. Babatola, D. Catoe, A. Boehm, M. Wilder, H. Green, A. Lobos, V. Harwood, S. Hertel, R. Klepikow, M. Howard, P. Laksanalamai, A. Roundtree, M. Mattioli, S. Eytcheson, M. Molina, M. Lane, R. Rediske, A. Ronan, N. D'Souza, J. Rose, A. Shrestha, C. Hoar, A. Silverman, W. Faulkner, K. Wickman, J. Kralj, S. Servetas, M. Hunter, S. Jackson, AND O. Shanks. Interlaboratory performance and quantitative PCR data acceptance metrics for NIST SRM® 2917. WATER RESEARCH. Elsevier Science Ltd, New York, NY, 225:119162, (2022). https://doi.org/10.1016/j.watres.2022.119162
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 multiple water safety 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:
Surface water quality quantitative polymerase chain reaction (qPCR) technologies are expanding from a subject of research to routine environmental and public health laboratory testing. Readily available, reliable reference material is needed to interpret qPCR measurements, particularly across laboratories. Standard Reference Material® 2917 (NIST SRM® 2917) is a DNA plasmid construct that functions with multiple water quality qPCR assays allowing for estimation of total fecal pollution and identification of key fecal sources. This study investigates SRM 2917 interlaboratory performance based on repeated measures of 12 qPCR assays by 14 laboratories (n = 1008 instrument runs). Using a Bayesian approach, single-instrument run data are combined to generate assay-specific global calibration models allowing for characterization of within- and between-lab variability. Comparable data sets generated by two additional laboratories are used to assess new SRM 2917 data acceptance metrics. SRM 2917 allows for reproducible single-instrument run calibration models across laboratories, regardless of qPCR assay. In addition, global models offer multiple data acceptance metric options that future users can employ to minimize variability, improve comparability of data across laboratories, and increase confidence in qPCR measurements.
URLs/Downloads:
DOI: Interlaboratory performance and quantitative PCR data acceptance metrics for NIST SRM® 2917
https://pubmed.ncbi.nlm.nih.gov/36191524/