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Also, the clinical relapse of disease in 4 APL patients with reverted PML/RAR alpha positivity is consistent with the view that detection of PML/RARalpha by RT-RCR in patients in remission implies a poor prognosis.
CNS involvement was diagnosed based on the appearance of APL blasts in cerebrospinal fluid (CSF).
After treated with all-trans retinoic acid (ATRA), idarubicin, cytarabine and intrathecal methotrexate, she achieved morphological, molecular, and CNS remission.
The typical translocation [t(15;17)(q22;q11)] was detected by cytogenetic analysis in 5 M3 patients, but PML/RARalpha was positive in 13 out of 15 patients, as assessed by RT-PCR (8 patients with bcr1 and 5 with bcr3 subtype).
Cytogenetic diversity was found in three patients (1 with t(17;17), 1 with 8, and 1 with add (7)(q22); -7; 8).
0.2% Triton X-100 and 0.2% saponin were used for the permeabilization step .
In hybridization, a wide range of times and temperatures were used and probe was designed in FRET system.
The goal of this study was to develop a new and powerful Flow- FISH technique to detect the long isoform (L) of PML-RARa fusion transcript in NB4 cell line.
METHODS: To achieve the best condition for fixation, two different fixatives including 2% paraformaldehyde and 75% ethanol were used.
Using standard fixation and permeabilization protocol of 2% PFA and 0.2% saponin gave the best fluorescent results in flow cytometry.
Also, results indicated that the optimum time and temperature for hybridization was 2 hr at 42°C.