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Effect of the Ratio of Free to Total Prostate-specific Antigen on Interassay Variability in Proficiency Test Samples

¹ Division of Molecular Medicine, Wadsworth Center, New York State Department of Health, Albany, NY 12201.
Departments of
² Biomedical Sciences and
³ Biometry and Statistics, School of Public Health, University at Albany, Albany, NY 12201.
^a Address correspondence to this author at: State of New York Department of Health, Wadsworth Center, Empire State Plaza, P.O. Box 509, Albany, NY 12201. Fax 518-473-2900; e-mail schneid{at}wadsworth.org

Background: Up to sevenfold differences were observed between total prostate-specific antigen (PSA) methods for New York State Proficiency Test samples prepared with seminal fluid PSA in human female serum. Because the PSA was mainly in its free form under these conditions, we wanted to determine whether a defined mixture of free and complexed PSA would reduce the interassay differences.

Methods: We prepared a series of five solutions of 60 g/L bovine serum albumin with 10 µg/L total PSA consisting of varied proportions of free, noncomplexible PSA, and ₁-antichymotrypsin (ACT)-complexed PSA from 0% to 100%. Two hundred seventy laboratories measured the total PSA in these samples, and 16 laboratories also analyzed the samples for free PSA. The results were used to calculate free/total PSA ratios.

Results: Interassay CVs for total PSA measurements were ~7% at 10–15% free PSA but became gradually larger as the free/total PSA ratio increased. Measured free-PSA concentrations were similar within each sample (mean CV, 12%), and the results were relatively independent of the proportion of free PSA in the samples. Twofold discrepancies between actual and expected ratios were observed with some methods at 100% free PSA and to a lesser degree at 30% free PSA. At 100% free PSA, the relatively higher total-PSA values measured by nonequimolar methods yielded low free/total PSA ratios of 50–60%. In contrast, the lower total PSA values obtained by equimolar methods yielded ratios close to the expected 100%.

Conclusions: Preparing proficiency test samples with a 10:90 mixture of free, noncomplexible PSA:PSA-ACT is a viable alternative to the use of seminal fluid PSA. Furthermore, the method used to measure total PSA may have a substantial impact on the calculated proportion of free PSA and hence may have clinical relevance.© 1999 American Association for Clinical Chemistry

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