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Comprehensive validation of a real-time quantitative bcr-abl assay for clinical laboratory use AMERICAN JOURNAL OF CLINICAL PATHOLOGY Jones, C. D., Yeung, C., Zehnder, J. L. 2003; 120 (1): 42-48

Abstract

We developed and extensively validated a real-time PCR assay for the quantitation of bcr-abl to determine residual disease in patients with chronic myelogenous leukemia. This method quantitates the p210 and the p190 bcr-abl RNA fusion transcripts with results normalized to a housekeeping gene, using the 5'-exonuclease technique and the ABI PRISM 7700 Sequence Detection System (Applied Biosystems, Foster City, CA). We parallel tested 372 clinical specimens and 50 peripheral blood samples from patients not known to have any myeloproliferative disorders. The results were 100% specific. Sensitivity studies showed that this method can detect bcr-abl in cell lines diluted to 0.0001% and can detect a single bcr-abl plasmid spiked into negative RNA. The between-run reproducibility showed a coefficient of variance (CV) of 12.3%, and within-run reproducibility showed a CV of 13.8%. This method can be used to reliably monitor the disease load in patients with bcr-abl-positive diseases.

View details for DOI 10.1309/60A9C8WGEGHRNXEE

View details for Web of Science ID 000183730300005

View details for PubMedID 12866371