Rapid detection of leukemia-associated translocation fusion genes using a novel combined RT-PCR and flow cytometric method.
Efficient detection of recurrent translocation-associated fusion genes is of critical importance for the diagnosis, prognosis and post-therapeutic monitoring of many leukemias. Typically, the presence of such translocations is revealed by RT-PCR technique, followed by Southern blot hybridization to ensure specificity of the PCR product. Though widely employed, post-PCR analysis of this type is relatively laborious and time-intensive. As a departure from standard analytic approaches, we have developed a robust novel method combining both high specificity and sensitivity, based on polystyrene bead capture of fluorescently labeled PCR products, with subsequent analysis by flow cytometry. Results from cell line and patient sample evaluations indicate that this method may be easily incorporated into the diagnostic molecular laboratory as a rapid and cost-effective alternative to currently employed techniques.
Zhang
Q-Y
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Department of Pathology, Experimental Pathology Laboratory, University of New Mexico Health Sciences Center, Albuquerque, NM, USA.
Garner
K
K
Viswanatha
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D000328:Adult; D019921:Biotinylation; D002648:Child; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics; D002888:Chromosomes, Human, Pair 19 / Q000235:genetics; D003362:Cost-Benefit Analysis; D004273:DNA, Neoplasm / Q000032:analysis* / Q000235:genetics; D005434:Flow Cytometry / Q000191:economics / Q000379:methods*; D005456:Fluorescent Dyes; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D020014:K562 Cells / Q000737:chemistry; D007938:Leukemia / Q000235:genetics* / Q000473:pathology; D008863:Microspheres; D015345:Oligonucleotide Probes; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D010677:Philadelphia Chromosome; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics; D012680:Sensitivity and Specificity; D013997:Time Factors; D014178:Translocation, Genetic / Q000235:genetics*; D014407:Tumor Cells, Cultured / Q000737:chemistry
D004273:DNA, Neoplasm; D005456:Fluorescent Dyes; D018398:Homeodomain Proteins; D015345:Oligonucleotide Probes; D015514:Oncogene Proteins, Fusion; D012333:RNA, Messenger; D012334:RNA, Neoplasm; C089847:E2A-Pbx1 fusion protein; D016044:Fusion Proteins, bcr-abl