Detection of prostatic specific membrane antigen messenger RNA using immunobead reverse transcriptase polymerase chain reaction Journal Article

  

Authors:	Ghossein, R. A.; Osman, I.; Bhattacharya, S.; Ferrara, J.; Fazzari, M.; Cordon-Cardo, C.; Scher, H. I.
Article Title:	Detection of prostatic specific membrane antigen messenger RNA using immunobead reverse transcriptase polymerase chain reaction
Abstract:	The present study was performed to detect circulating prostatic carcinoma (PC) cells using a novel three-step immunobead reverse transcriptase (RT) polymerase chain reaction (PCR) assay for prostatic specific membrane antigen (PSMA) messenger RNA (mRNA). The sensitivity and specificity of this technique was assessed and the incidence of immunobead RT-PCR positivity correlated with progressive metastatic disease and serum prostatic specific antigen (PSA) levels. Fifty peripheral blood (PB) samples from 46 patients with PC were incubated with magnetic beads coated with Ber- EP4 antibody directed against the human epithelial antigen, a membrane antigen widely expressed by epithelial cells. The epithelial cell-enriched magnetic fraction was then subjected to mRNA isolation using oligo- deoxythymidine (dT) magnetic beads. Nested RT-PCR for PSMA was performed on the mRNA oligo-dT complex and the identity of the RT-PCR products was confirmed by Southern blotting. Twenty-one PB samples from 8 control subjects without PC were also evaluated. Three-step immunobead PSMA RT-PCR was able to detect one PC cell per 1 mL of PB. The positivity rate of the RT-PCR assay was significantly higher (11 of 25; 44%) in patients with metastatic tumor than in patients with non-metastatic disease (1 of 21; 5%) (P = 0.003). In patients with metastatic PC, RT-PCR positivity was much higher in patients with progressive disease (10 of 13; 77%) than in patients with responding or stable disease (1 of 12; 8%) (P = 0.001). There was a statistically significant correlation between immunobead PSMA PCR positivity and high levels of serum PSA (P = 0.005). All control subjects without PC tested negative for PSMA PCR. The three-step immunobead RT-PCR for PSMA can detect circulating PC cells with high specificity and sensitivity. Preliminary data show a strong correlation between immunobead PCR positivity, the presence of progressive metastatic disease, and high levels of serum PSA.
Keywords:	clinical article; controlled study; sensitivity and specificity; antigen expression; prostate specific antigen; metastasis; reverse transcription polymerase chain reaction; tumor markers, biological; tumor cells, cultured; prostate-specific antigen; prostatic neoplasms; messenger rna; reverse transcriptase polymerase chain reaction; rna, messenger; antigen detection; nucleotide sequence; immunomagnetic separation; antibody; carcinoma cell; glutamate carboxypeptidase ii; antigens, surface; prostate carcinoma; epithelial membrane antigen; southern blotting; gene isolation; thymidine derivative; neoplasm circulating cells; blotting, southern; prostatic carcinoma; humans; human; male; priority journal; article; electrophoresis, agar gel; carboxypeptidases; immunobead; prostatic specific membrane antigen
Journal Title:	Diagnostic Molecular Pathology
Volume:	8
Issue:	2
ISSN:	1052-9551
Publisher:	Lippincott Williams & Wilkins
Date Published:	1999-06-01
Start Page:	59
End Page:	65
Language:	English
DOI:	10.1097/00019606-199906000-00001
PUBMED:	10475379
PROVIDER:	scopus
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-0032795596&partnerID=40&md5=4cfa16a2ca6e37249285c1f886c7d2e4
Notes:	Article -- Export Date: 16 August 2016 -- Source: Scopus

https://synapse.mskcc.org/synapse/works/has_related_data_chk/96150