Automated agglutination analyzer with contour comparison
Inventors
Rowlen, Kathy L. • WILSON, Garrett • YAMADA, Gregory • Smolak, Andrew • Rauker, Goran • Ives, Jeffrey T.
Assignees
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Publication Number
US-10948420-B2
Publication Date
2021-03-16
Expiration Date
Abstract
The systems and methods contained herein are directed toward automated analysis of agglutination reactions to determine properties of materials, including viruses and vaccines thereto. Advanced digital imaging and processing techniques are used to determine the presence or absence of viruses or antibodies within a fluid sample. The systems and methods are versatile, and can be used to determine specific properties of biomaterials and viruses, such as titer value, concentration, genotype, phenotype, serotype, vaccine efficacy, viral resistance and other properties of relevance in the medical, research and development fields. Also provided are systems and methods of standardization, repeatability, and data storage and transmittal to reduce errors and subjectivity inherent to conventional assays characterized by human readers.
Core Innovation
The invention relates to an automated agglutination analyzer that determines an agglutination parameter using digital imaging and image-processing of a reaction plate having a plurality of reaction vessels. The method spans a dilution range of an introduced agglutinating mediator across reaction vessels, creates a digital image of the vessels, and processes the digital image by defining areas corresponding to each reaction vessel and associating the areas with pixels for subsequent measurement.
The method measures pixel intensity within each defined area, identifies a boundary contour for each reaction vessel when present, and calculates one or more boundary contour parameters for each identified boundary contour. The calculated boundary contour parameters for each reaction vessel are compared to one or more standard boundary contour parameters from a control reaction vessel having a non-agglutination control or an agglutination control, and an agglutination condition is determined for each reaction vessel as agglutination, non-agglutination, non-specific inhibition, or error state.
An agglutination parameter is then determined from the agglutination condition that is agglutination or non-agglutination in at least a portion of the reaction vessels over the dilution range, and the agglutination parameter is defined as titer or concentration of the agglutinating mediator. The invention also provides a method for determining validation status of an automated agglutination analyzer by imaging a quality control plate with a first set of wells having an optically dense button and a second set without the optically dense button, and comparing boundary contour parameters to standard parameters corresponding to non-agglutination versus agglutination.
Claims Coverage
The partial content identifies two independent claims. Across these, the inventive features cover digital-image boundary contour parameter measurement and comparison for determining an agglutination condition and deriving an agglutination parameter over a dilution range, and using a QC plate with an optically dense button/no-button well pattern and boundary contour parameter comparison to determine validation status of an automated agglutination analyzer.
Digital image processing with boundary contour parameters and control comparison
A method comprising providing a reaction plate having a plurality of reaction vessels, introducing an agglutinating mediator spanning a dilution range, introducing particles, creating a digital image, processing the image by defining vessel-corresponding areas and pixels, measuring pixel intensity, identifying a boundary contour for each vessel from the measuring, calculating one or more boundary contour parameters, and comparing calculated boundary contour parameters in each vessel with one or more standard boundary contour parameters from a control reaction vessel having a non-agglutination control or an agglutination control to determine an agglutination condition that is agglutination, non-agglutination, non-specific inhibition, or error state.
Determining an agglutination parameter from agglutination condition over a dilution range
The agglutination condition determined as agglutination or non-agglutination in at least a portion of the plurality of reaction vessels over the dilution range is used to determine an agglutination parameter, wherein the agglutination parameter is titer or concentration of the agglutinating mediator.
Validation status determination using a QC plate with optically dense button wells
A method for determining validation status of an automated agglutination analyzer by providing a quality control plate with a plurality of wells including a first set having an optically dense button and a second set without the optically dense button, creating a digital image, processing the image by defining areas corresponding to each well, measuring pixel intensity within each defined area, identifying a boundary contour if present, calculating one or more boundary contour parameters for each identified boundary contour, and comparing calculated boundary contour parameters with one or more standard boundary contour parameters corresponding to non-agglutination or agglutination to determine validation status.
Validated and not validated outcomes based on expected optically dense button correspondence
Validation is determined by the comparing step resulting in wells having the optically dense button corresponding to non-agglutination and wells without the optically dense button corresponding to agglutination; otherwise the automated agglutination analyzer is not validated when the comparing step results in at least one reaction vessel that is not non-agglutination for wells having the optically dense button and that is not agglutination for wells not having the optically dense button.
Overall, the independent claims cover an image-processing-based classification workflow using boundary contour parameters compared against standard parameters from control to assign agglutination conditions and derive an agglutination parameter across a dilution range, and a separate QC-based boundary contour parameter comparison approach to determine whether the automated agglutination analyzer is validated or not validated using optically dense button versus no-button wells.
Stated Advantages
Enables determination of an agglutination parameter (titer or concentration) from agglutination conditions assigned to reaction vessels over a dilution range.
Provides a validation status determination for an automated agglutination analyzer using a QC plate with optically dense button and without-button wells.
Documented Applications
Hemagglutination (HA) and hemagglutination inhibition (HI) contexts are described as example workflows using the automated agglutination analyzer.
Instrument verification and algorithm/lab/operator verification are described using a quality control plate with optically dense button versus no-button wells.
Determining validation status of an automated agglutination analyzer is described using boundary contour parameter comparison to standard parameters from non-agglutination versus agglutination.
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