Modified assay of prostate-specific antigen with a detection limit < 0.01 microgram/L

We modified the Hybritech Tandem-E prostate-specific antigen (PSA) assay by increasing the sample volume, increasing enzyme-substrate incubation time, and using diethanolamine buffer. Our modified method has a detection limit of 0.009 microgram/L (P < 0.01). The assay curve is linear from 0.01 to 1.0 micrograms/L and has an overall assay time of about 4 h. Linear plots are obtained when the 1.0 micrograms/L standard is diluted with either matrix buffer or serum from men containing PSA < 0.01 microgram/L. Recovery of PSA (0.10 microgram/L) added to serum from men averaged 94%. Interassay CVs were 13%, 7%, and 4% at PSA concentrations of 0.04, 0.07, and 0.30 micrograms/L, respectively (n = 33). This assay should be useful in the detection of early recurrence of prostate cancer after radical prostatectomy.