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A quantitative reverse transcriptase polymerase chain reaction method for the detection of leukaemic cells with t(8;21) in peripheral blood

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Abstract: We evaluated the usefulness of a recently developed real‐time reverse transcriptase polymerase chain reaction (RT‐PCR) system to detect minimal residual diseases (MRD) in patients with acute myelogenous leukaemia (AML) with chromosomal translocation t(8:21). The method was simple, rapid and reproducible for the quantity of chimeric AML1‐ETO (MTG8) transcripts. The ratio of the absolute copy number of a target gene (AML1‐ETO) to a control gene (glyceraldehyde‐3‐phosphate dehydrogenase, GAPDH) was calculated by using a fluorescence curve prepared from amplicons of serially diluted standard RNA. The relative points of MRD in bone marrow (BM) of 8 patients in the acute phase of the disease was from 0.85 to 3.0, whereas those of MRD in complete remission (CR) decreased to below 6.4×10−3. This method was also applied to evaluate chimeric transcripts in peripheral blood (PB) samples. The values in patients with t(8;21) AML were from 0.97 to 2.0 in the acute phase, whereas those in CR showed less than 2.2×10−4. There was 10−5‐fold difference in AML1‐ETO mRNA expression between PB samples in the acute phase and those in CR. The results suggest that we may easily monitor MRD in patients with t(8;21) AML through quantitative analysis of AML1‐ETO transcripts in blood samples.
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Keywords: AML1‐ETO (MTG8); RT‐PCR; acute myelogenous leukaemia (AML); minimal residual disease (MRD); real‐time quantitative PCR system; t(8;21)

Document Type: Research Article

Affiliations: 1: Department of Clinical and Laboratory Medicine, 2: Department of Pediatrics, Tohoku University, School of Medicine, Sendai, Japan; and 3: Department of Cancer Chemotherapy, Saitama Cancer Center, Ino, Japan 4: Department of Information Science, and

Publication date: April 1, 2000

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