Understanding Variance in Multiplex Suspension Array Assays
Author Information
Author(s): Hanley Brian P
Primary Institution: University of California, Davis
Hypothesis
It would be practical to obtain 1,000 microspheres for each of the 7 microsphere classifiers in the assay for all of the sample wells.
Conclusion
Increasing the number of microspheres acquired per classifier does not effectively address the issues of reliability or improved precision of these assays in most situations.
Supporting Evidence
- The study attempted to replicate previous results showing that larger numbers of microspheres could increase accuracy.
- Significant variation was found in microsphere counts across different wells.
- The overall mean number of microspheres acquired was 1521 with a standard deviation of 299.
Takeaway
This study looked at how many tiny beads are needed in tests to get good results, and found that just adding more beads doesn't always help.
Methodology
The study used a 7-plex multiplexed assay with 10,000 microspheres per classifier injected into 17 replicate wells.
Potential Biases
Potential aggregation of microspheres and preferential binding to wells could introduce bias.
Limitations
The study does not present data on relative confidence intervals or standard error obtainable using larger than normal numbers of microspheres per classifier.
Participant Demographics
Highly characterized serum from a single Rhesus macaque was utilized.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website