Interlaboratory performance and quantitative PCR data acceptance metrics for NIST SRM® 2917.

Area of Science:

• Environmental Science
• Analytical Chemistry
• Molecular Biology

Background:

• Quantitative polymerase chain reaction (qPCR) is increasingly used for water quality testing.
• Reliable reference materials are crucial for accurate and comparable qPCR results across laboratories.
• Standard Reference Material® 2917 (NIST SRM® 2917) is a DNA plasmid designed for water quality qPCR assays.

Purpose of the Study:

• To evaluate the interlaboratory performance of NIST SRM® 2917.
• To assess the utility of SRM® 2917 for establishing reproducible calibration models.
• To develop data acceptance metrics for new SRM® 2917 users.

Main Methods:

• A Bayesian approach was used to combine data from 14 laboratories and 12 qPCR assays (n=1008 runs).
• Interlaboratory performance was assessed through repeated measures of SRM® 2917.
• Additional data sets were used to validate new data acceptance metrics.

Main Results:

• SRM® 2917 demonstrated reproducible single-instrument run calibration models across diverse laboratories and qPCR assays.
• Global calibration models effectively characterized within- and between-laboratory variability.
• The study proposed multiple data acceptance metric options for users.

Conclusions:

• NIST SRM® 2917 facilitates reproducible qPCR calibration models for water quality testing.
• The use of SRM® 2917 and proposed metrics can minimize variability and enhance data comparability.
• Implementing SRM® 2917 increases confidence in environmental and public health qPCR measurements.