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Clinical Chemistry 52: 1089-1095, 2006. First published April 20, 2006; 10.1373/clinchem.2005.063289
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(Clinical Chemistry. 2006;52:1089-1095.)
© 2006 American Association for Clinical Chemistry, Inc.

Cancer Diagnostics

APTIMA PCA3 Molecular Urine Test: Development of a Method to Aid in the Diagnosis of Prostate Cancer

Jack Groskopf1,a, Sheila M.J. Aubin1, Ina Lim Deras1, Amy Blase1, Sharon Bodrug1, Craig Clark1, Steven Brentano1, Jeannette Mathis1, Jimmykim Pham1, Troels Meyer1, Michelle Cass1, Petrea Hodge1, Maria Luz Macairan2, Leonard S. Marks2 and Harry Rittenhouse1

1 Gen-Probe Incorporated, San Diego, CA.
2 Urological Sciences Research Foundation, Culver City, CA.

aAddress correspondence to this author at: Gen-Probe Incorporated, 10210 Genetic Center Dr., San Diego, CA 92121. Fax 858-410-8113; e-mail jackg{at}gen-probe.com.

Background: Prostate cancer gene 3 (PCA3) encodes a prostate-specific mRNA that has shown promise as a prostate cancer diagnostic tool. This report describes the characterization of a prototype quantitative PCA3-based test for whole urine.

Methods: Whole-urine specimens were collected after digital rectal examination from 3 groups: men scheduled for prostate biopsy (n = 70), healthy men (<45 years of age with no known prostate cancer risk factors; n = 52), and men who had undergone radical prostatectomy (n = 21). PCA3 and prostate-specific antigen (PSA) mRNAs were isolated, amplified, and quantified by use of Gen-Probe DTS400® Systems. Prostate biopsy results were correlated with the PCA3/PSA mRNA ratio, and PSA mRNA concentrations were used to normalize PCA3 signals and confirm the yield of prostate-specific RNA. Assay precision, specimen stability, and mRNA yield were also evaluated.

Results: The specimen informative rate (fraction of specimens yielding sufficient RNA for analysis) was 98.2%. In this clinical research study, ROC curve analysis of prebiopsy specimens yielded an area under the curve of 0.746; sensitivity was 69% and specificity 79%. Serum PSA assay specificity was 28% for this same group. PCA3 and PSA mRNAs were undetectable in postprostatectomy specimens except for one man with recurrent prostate cancer. Assay interrun CVs were ≤12%. Both mRNAs were stable in processed urine up to 5 days at 4 °C and after 5 freeze–thaw cycles.

Conclusion: The APTIMA® PCA3 assay combines simple specimen processing with precise assays and existing instruments and could add specificity to the current algorithm for prostate cancer diagnosis.




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