Determination of immunoglobulin heavy variable (IGHV) gene mutational status in chronic lymphocytic leukemia
Chronic lymphocytic leukemia (CLL) is the most common type of adult leukemia in Western countries, accounting for 30% of all leukemias, and manifests as clonal expansion of mature CD5+ CD19+ CD23+ sIgMlow B lymphocytes. Among novel molecular markers associated with clinical course and prognosis of CLL, mutational status of immunoglobulin heavy variable (IGHV) genes turned out to be of the greatest prognostic value as the most informative and most stable marker which can be assessed at any time of the disease. IGHV mutational status defines two CLL subsets; the patients expressing unmutated IGHV genes usually suffer from more agressive disease and have worse prognosis compared to patients with mutated IGHV genes.
Starting material for this analysis is RNA isolated from peripheral blood mononuclear cells. IGHV mutational status is being determined by RT-PCR amplification of clonal rearrangement followed by direct sequencing.
It is necessary to deliver 10 ml of peripheral blood, using 3.8% sodium citrate as anticoagulant in vol/vol ratio 9:1. The sample is required to be delivered fresh, without freezing, during the same day. The samples are accepted every day from 10 to 13h. It is possible to deliver the sample via post express service. The analysis is performed within 10 working days. The results and the bill are sent to the patient's home address, or can be collected personally at the Institute from 10 to 16h every day.
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