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Detection of Germ-Cell Tumor Cells in the Peripheral Blood by Nested Reverse Transcription-Polymerase Chain Reaction for -Fetoprotein-Messenger RNA and ß Human Chorionic Gonadotropin-Messenger RNA¹

Department of Urology [A. L. A. H., H. M., A. B., H. R.], Institute of Virology [M. L., M. R.], and Department of Medicine, Tumor Research [A. H.] University of Essen, D-45122 Essen, Germany

By establishing sensitive nested reverse transcription-PCRs for the detection of mRNA of -fetoprotein (AFP) and ß human chorionic gonadotropin (ßhCG), we investigated the presence of circulating tumor cells in the peripheral blood of 119 patients with germ-cell tumor. A total of 336 blood samples obtained before and during therapy were examined with regard to clinical applicability. The overall ratio of positive PCR results was 26.5% and was independent of the serum concentration of AFP and hCG/ßhCG. No correlation of the positivity for AFP-mRNA to serum AFP level was found. In contrast, positive results in ßhCG-PCR were twice as frequent in patients with elevated serum hCG/ßhCG levels as in those with normal serum hCG/ßhCG levels (P = 0.012). To develop a valid correlation to tumor stage, tumor histology, and serum level of tumor markers, a subgroup of 36 patients was evaluated before definite therapy. The subgroup revealed an overall ratio of 33.3% positive PCR results. The serum level of both of the markers did not correlate with the detection of corresponding mRNA in peripheral blood samples. However, positive ßhCG-PCR results were found exclusively in patients with elevated serum hCG/ßhCG (6 of 18 versus 0 of 18; P = 0.019). Patients with stage IIC/III germ-cell tumor demonstrated nearly twice the frequency of positive PCR results as patients with stage I tumor [7 (41.2%) of 17 versus 4 (23.5%) of 17] in this subgroup. With regard to histology, positive PCR results were found mostly in embryonal carcinoma.

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