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Comparison of the RNA-amplification based methods RT-PCR and NASBA for the detection of circulating tumour cells.

Burchill SA, Perebolte L, Johnston C, Top B, Selby P - Br. J. Cancer (2002)

Bottom Line: This may result in a redefinition of disease-free and clinical relapse.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.This may limit the routine use of NASBA for the detection of clinically significant disease.

View Article: PubMed Central - PubMed

Affiliation: Children's Cancer Research Laboratory, St. James's University Hospital, Leeds LS9 7TF, UK. s.a.burchill@leeds.ac.uk

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.

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Related in: MedlinePlus

Schematic representation of methods for the amplification of mRNA using (A) NASBA, and (B) RT–PCR.
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fig1: Schematic representation of methods for the amplification of mRNA using (A) NASBA, and (B) RT–PCR.

Mentions: Increasingly, reverse transcriptase polymerase chain reaction (RT–PCR) (Figure 1AFigure 1


Comparison of the RNA-amplification based methods RT-PCR and NASBA for the detection of circulating tumour cells.

Burchill SA, Perebolte L, Johnston C, Top B, Selby P - Br. J. Cancer (2002)

Schematic representation of methods for the amplification of mRNA using (A) NASBA, and (B) RT–PCR.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2746547&req=5

fig1: Schematic representation of methods for the amplification of mRNA using (A) NASBA, and (B) RT–PCR.
Mentions: Increasingly, reverse transcriptase polymerase chain reaction (RT–PCR) (Figure 1AFigure 1

Bottom Line: This may result in a redefinition of disease-free and clinical relapse.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.This may limit the routine use of NASBA for the detection of clinically significant disease.

View Article: PubMed Central - PubMed

Affiliation: Children's Cancer Research Laboratory, St. James's University Hospital, Leeds LS9 7TF, UK. s.a.burchill@leeds.ac.uk

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.

Show MeSH
Related in: MedlinePlus