We used a recently developed system for real-time quantitative polymerase chain reaction (PCR) to determine @entity66 in @entity1 with @entity1860 . The expression of the @entity1861 hybrid oncogene was determined and normalized by using the @entity6049 housekeeping gene product as endogenous reference. The sensitivity and reproducibility of the assay was tested on cell line K562. A dilution of @entity1861 -positive cell line K562 remained positive at up to 250 fg of RNA. 10 copies of @entity1861 DNA in water could still be detected. The dynamic range of the method spanned six orders of magnitude. Analysis of 10 identical assays on K562 RNA resulted in a variation of 15%. To test the feasibility of normalization of @entity1861 dosage by the @entity6049 product, we compared the efficiencies of the RT-PCRs in 150 @entity1 analyses. We concluded that @entity6049 was a suitable and stringent quality control standard. Three @entity1 who were treated with donor leucocyte infusions for @entity1860 who had relapsed after bone marrow transplantation were followed over time. The normalized @entity1861 dosage was compared to the results of cytogenetics. Cytogenetic analysis was negative below a normalized @entity1861 dose of about 3 x 10(-2). This semi-automated method is fast, sensitive and accurate and enables a high throughput of samples.

Quantitation of minimal @entity66 in XXXX @entity1 using real-time quantitative RT-PCR.

@entity1 :: ('9606', 'Species') :: ['patients', 'patient'] @entity1861 :: ('25', 'Gene') :: ['Bcr-Abl'] @entity66 :: ('MESH:D004194', 'Disease') :: ['residual disease'] @entity6049 :: ('3145', 'Gene') :: ['PBGD'] @entity1860 :: ('MESH:D015464', 'Disease') :: ['chronic myeloid leukaemia']
@entity1860 :: (MESH:D015464,Disease) :: ['Philadelphia chromosome positive chronic myeloid leukaemia']
