title: Comparison of the RNA-amplification based methods RT-PCR and NASBA for the detection of circulating tumour cells.
abstract: Increasingly, reverse transcriptase polymerase chain reaction (RT-PCR) is used to detect clinically significant tumour cells in blood or bone marrow. This may result in a redefinition of disease-free and clinical relapse. However, its clinical utility may be limited by lack of automation or reproducibility. Recent studies have suggested nucleic acid sequence-based amplification of target RNA may be more robust. In this study, nucleic acid sequence-based amplification was established to detect melanoma, colorectal and prostate cancer cells. Nucleic acid sequence-based amplification and RT-PCR both successfully amplified target RNA in peripheral blood samples from patients with melanoma and colorectal cancer, but only RT-PCR detected PSA in blood samples from patients with prostate cancer. There was relatively good agreement between sample replicates analyzed by RT-PCR (Kappa values of one for tyrosinase, 0.67 for CK-20 and one for PSA), but less agreement when analyzed by nucleic acid sequence-based amplification. This may limit the routine use of NASBA for the detection of clinically significant disease. In summary, RT-PCR appears at present to be the most reliable and reproducible method for the detection of low-level disease in cancer patients, although prospective studies are warranted to assess the clinical utility of different molecular diagnostic methods.
authors.affiliation: Children's Cancer Research Laboratory, St. James's University Hospital, Leeds LS9 7TF, UK. s.a.burchill@leeds.ac.uk
authors.forename: S A
authors.initials: SA
authors.lastname: Burchill
authors.forename: L
authors.initials: L
authors.lastname: Perebolte
authors.forename: C
authors.initials: C
authors.lastname: Johnston
authors.forename: B
authors.initials: B
authors.lastname: Top
authors.forename: P
authors.initials: P
authors.lastname: Selby
mesh_terms: D015179:Colorectal Neoplasms / Q000175:diagnosis*; D003402:Creatine Kinase / Q000235:genetics; D005784:Gene Amplification; D006801:Humans; D008297:Male; D008545:Melanoma / Q000175:diagnosis*; D014442:Monophenol Monooxygenase / Q000235:genetics; D009360:Neoplastic Cells, Circulating; D017430:Prostate-Specific Antigen / Q000235:genetics; D011471:Prostatic Neoplasms / Q000175:diagnosis*; D012333:RNA, Messenger / Q000032:analysis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: D012333:RNA, Messenger; D014442:Monophenol Monooxygenase; D003402:Creatine Kinase; D017430:Prostate-Specific Antigen