2012
DOI: 10.1016/j.cancergen.2012.06.006
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An optimized technology platform for the rapid multiplex molecular analysis of genetic alterations associated with leukemia

Abstract: Molecular methods play a critical role in the accurate diagnosis of leukemia by complementing morphologic, cytochemical, immunophenotypic, and cytogenetic analyses. We developed a multiplex RT-PCR method combined with liquid bead array cytometry for the rapid detection of genetic alterations associated with leukemia. Fusion transcripts corresponding to the most common recurrent chromosomal translocations were reproducibly detected in as low as 0.1 to 10 ng of total RNA with an analytical sensitivity of 0.01 to… Show more

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Cited by 6 publications
(7 citation statements)
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“…A DNA Engine Tetrad 2 (Bio-Rad, Hercules, CA, USA) was used at site 2. The protocols for the modified prototype panels are described in Ye et al 17 …”
Section: Methodsmentioning
confidence: 99%
“…A DNA Engine Tetrad 2 (Bio-Rad, Hercules, CA, USA) was used at site 2. The protocols for the modified prototype panels are described in Ye et al 17 …”
Section: Methodsmentioning
confidence: 99%
“…Total nucleic acid (TNA) was extracted from RNARetain-preserved FNAs using a proprietary laboratory-validated method and quantified using a NanoDrop ND1000 (NanoDrop Technologies, Waltham, MA). DNA-and RNA-based tests were performed using 20 or 40 ng of purified TNA, a single qualitative multiplex assay system based on the Signature technology platform (17)(18)(19) and reagents developed, manufactured, qualified, and released under controlled processes by Asuragen Inc., Austin, Texas. To generate qualitative positive or negative test results, assay outputs (median fluorescence intensity signals) were interpreted relative to target-specific, predetermined, laboratory-validated qualitative cutoff values corresponding to an analytical sensitivity of 0.5% to 2.5%.…”
Section: Molecular Testingmentioning
confidence: 99%
“…The reference diagnosis was established using local pathology expertise while molecular testing was performed using standardized molecular methods at a centralized clinical laboratory (17)(18)(19). All cytologic, molecular, and histologic results were collected and analyzed, including aspirates with nondiagnostic cytopathology.…”
Section: Introductionmentioning
confidence: 99%
“…15,16 Likewise, solution hybridization of multiple PCR products on addressable microspheres, followed by flow cytometry fluorescence analysis, is broadly used in clinical diagnostic applications for bacterial, viral, genetic, or oncology targets. [17][18][19][20][21] Regardless of the technology platform, molecular methods currently in clinical use have been optimized to be compatible with anatomic pathology specimens. Because of the breadth of specimen types and preanalytic parameters commonly encountered in the clinical setting, the quantity or quality of nucleic acids recovered from these specimens can vary greatly.…”
mentioning
confidence: 99%