Troubleshooting immunoassays

Immunoassay troubleshooting is a sequential, cumulative process whereby one first looks at the calibration curve, then QC s, and finally the sample results. If the curve is not acceptable, the assay fails. If the curve is acceptable, then one reviews QC performance if QC performance is unacceptable, the assay fails. If QCs are acceptable, then one examines individual sample results. If the assay fails as the result of an apparent technical error, the error is corrected and the assay is repeated. If the cause for the failure is not apparent or the assay has failed consistently, then a troubleshooting process should be initiated prior to further analysis (and depletion) of study samples. A summary of the calibrator/QC/sample result troubleshooting process is depicted in Fig. 3.14. 

B. D. Hammock, Troubleshooting during the development and use of immunoassays for environmental monitoring, in New Frontiers in Agrochemical Immunoassay, ed. D. A. Kurtz,... 

Wu JT. Quantitative Immunoassay A practical guide for assay establishment, troubleshooting, and clinical application. Washington D.C AACC Press, 2000. 

Immunoassays developed along the recommendations provided above should perform consistently and generate reliable data over time. However, occasional failures happen variations in laboratory environmental conditions, reagent quality, operator skill, and instalment performance, for example, make systematic troubleshooting skills invaluable in discerning the cause for assay failures. 

Wu, J.T. (2000) Quantitative Immunoassay A Practical Guide for Assay Establishment, Troubleshooting, and Clinical Applications. AACC Press, Washington, DC. 

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