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ARS Home » Pacific West Area » Salinas, California » Crop Improvement and Protection Research » Research » Publications at this Location » Publication #419144
Research Project: Disease Management and Improved Detection Systems for Control of Pathogens of Vegetables and Strawberries

Location: Crop Improvement and Protection Research

Title: How controls improve diagnostic assay performance: Hitchhiker's guide to diagnostic assay controls

Author
item Martin, Frank
item BATUMAN, OZGUR - Pennsylvania State University
item Luster, Douglas
item MILES, TIMOTHY - Michigan State University
item RIVERA, YAZMIN - Animal And Plant Health Inspection Service (APHIS)
item SHARMA, POONAM - Oklahoma State University
item GEISER, DAVID - Pennsylvania State University
item CARDWELL, KITTY - Oklahoma State University

Submitted to: PhytoFrontiers
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/8/2025
Publication Date: 5/9/2025
Citation: Martin, F.N., Batuman, O., Luster, D.G., Miles, T., Rivera, Y., Sharma, P., Geiser, D., Cardwell, K. 2025. How controls improve diagnostic assay performance: Hitchhiker's guide to diagnostic assay controls. PhytoFrontiers. 5(2):127-137. https://doi.org/10.1094/PHYTOFR-10-24-0118-FI.
DOI: https://doi.org/10.1094/PHYTOFR-10-24-0118-FI

Interpretive Summary: This manuscript discusses the important of including control reactions in all diagnostic assays and provides rationale behind their use. The reader is also directed to a website where recommendations on the types of controls that could be used are listed.

Technical Abstract: Controls in diagnostic assays are included to provide important information about the accuracy of results the assay provides and reduces the incidence of false negatives. All diagnostic assays should include controls to confirm the assay is performing properly and account for the type of assay being used (immune-, molecular, and sequence based) as well as differences in efficiency of sample extraction from different matrices and final sample purity. For example, when sampling plant material, selecting a plant target for a control (termed endogenous control) is useful for confirming if the sample is pure enough to provide an accurate result, thereby preventing false negatives. For environmental samples, adding a known control target (termed exogenous control) can provide a reference for the ability of the sample to provide an accurate result. This type of exogenous control can also be useful to evaluate amplification efficiency in PCR based assays to ensure it was not impacted when quantification of the pathogen is the goal. Specific examples of types of controls for immuno-, molecular-, and sequence-based detection methods are discussed below along with limitations of their use and considerations to keep in mind when using them.