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CML Primer Continued
Genetic Tools Of CML Diagnosis
See More Details at CML Bouys
There are 3 complementary methods to diagnose, exclude and monitor CML status, the details of these 3 testing procedures is given in the Testing Procedures Article.
1. Cytogenetics
From bone marrow aspirate or biopsy, a cytogenetic analysis is done which counts the percentage of Ph chromosome present in 20-30 marrow cells.
2. FISH
From marrow or peripheral blood, average 200 cells are sampled by fluorescent probes to detect the BCR-ABL hybrid gene. The use of interphase cells and greater sampling size make it more sensitive than cytogenetic analysis. FISH also counts the percentage of Ph chromosomes in the blood or marrow.
3. RT-PCR
Reverse transcriptase polymerase chain reaction (RT-PCR) is used to monitor minimal residual disease and offers the greatest sensitivity of all testing procedures, being capable of detecting up to 1 bad cell in 1 million white blood cells. A qualitative RT-PCR can give simple test information about breakpoints and the degree of remission whereas a quantitative RT-PCR can monitor minimal residual disease. For RT-PCR, RNA is isolated from blood or marrow samples and reverse transcribed into its copy, cDNA. The cDNA is then subjected to PCR amplification using primers specific for either the major or minor breakpoints.
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