Development of an Integrated Assay for Detection of BCR-ABL RNA

Winn-Deen, Emily S.; Helton, Bret; Atta, Reuel Van; Wong, William G.; Peralta, Jeffrey; Wang, James; Tsongalis, Gregory J.; Belloni, Dorothy R.; Chan, David; Eshleman, James R.; Gocke, Christopher D.; Jobbágy, Zsolt; Beppu, Lan; Radich, Jerald P.

doi:10.1373/clinchem.2007.085472

Cited by 48 publications

(36 citation statements)

References 24 publications

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“…14 The GeneXpert system showed high sensitivity (91.2%) and specificity (95%) as compared with the clinical ASR reference method, based on the simplified result reporting (positive, negative, or invalid) of the system software. The few false-negative samples either yielded no BCR-ABL signal (BCR-ABL Ct ϭ 0) or were interpreted as negative by the software due to the preset threshold (BCR-ABL Ct must be less than or equal to 32 14 ). Further studies may be needed to fine tune criteria for reporting results as negative.…”

Section: Discussionmentioning

confidence: 94%

Evaluation of the Cepheid GeneXpert BCR-ABL Assay

Jobbágy

Atta²,

Murphy

et al. 2007

The Journal of Molecular Diagnostics

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Patients with chronic myeloid leukemia harbor the chromosomal translocation t(9;22), which corresponds to fusion of the BCR and ABL genes at the DNA level. The translated fusion product is an oncogenic protein with increased ABL tyrosine kinase activity causing cell transformation. To date, reverse transcriptase-polymerase chain reaction is considered the most sensitive method available for detecting low copy numbers of the BCR-ABL gene fusion. Recently, Cepheid introduced its GeneXpert-based assay for the identification of the BCR-ABL gene fusion in cells from blood samples. This system comprises a walkaway self-contained instrument that combines cartridge-based microfluidic sample preparation with reverse transcriptase-polymerase chain reaction-based fluorescent signal detection and BCR-ABL and ABL Ct (threshold cycle) determination. The difference between the BCR-ABL Ct and ABL Ct (⌬Ct) is expected to represent the ratio of the two populations of mRNAs and ultimately the percentage of neoplastic cells present. We tested whether this BCR-ABL fusion detection system could be used as a clinical diagnostic tool for monitoring patients with minimal residual disease of chronic myelogenous leukemia. We report similar performance characteristics, including limit of detection, specificity, sensitivity, and precision, of this automated BCR-ABL fusion detection system to those of a manual TaqMan reverse transcriptase-polymerase chain reaction-based test.

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Section: Discussionmentioning

confidence: 94%

Evaluation of the Cepheid GeneXpert BCR-ABL Assay

Jobbágy

Atta²,

Murphy

et al. 2007

The Journal of Molecular Diagnostics

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“…It therefore preserves the stability and homogeneity of the calibrators, 2 important characteristics for an analytical reference material. In our study, the direct heat-lysis protocol was compatible with multiplex and singleplex methods, 1-and 2-step protocols, various real-time PCR instruments from multiple manufacturers, manual and automated setups, and commercial and laboratorydeveloped reagents (2, 22, 26 -29 ), but it could not be evaluated with integrated closed assay systems such as the GenXpert instrument (31 ).…”

Section: Synthetic Bcr-abl1 Calibrator Panels For Is Standardizationmentioning

confidence: 95%

Establishment and Validation of Analytical Reference Panels for the Standardization of Quantitative BCR-ABL1 Measurements on the International Scale

et al. 2013

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BACKGROUND: Current guidelines for managing Philadelphia-positive chronic myeloid leukemia include monitoring the expression of the BCR-ABL1 (breakpoint cluster region/c-abl oncogene 1, nonreceptor tyrosine kinase) fusion gene by quantitative reverse-transcription PCR (RT-qPCR). Our goal was to establish and validate reference panels to mitigate the interlaboratory imprecision of quantitative BCR-ABL1 measurements and to facilitate global standardization on the international scale (IS).

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“…185 It is likely that some of the intrinsic difficulties related to the complex time consuming methodology which requires specialized skills and knowledge may be overcome by the new automated BCR-ABL1 assay that is contained within a single-use microfluidic cartridge, using a specialized instrument, such as the Cepheid GeneXpert. 186 It is of interest that this specialized equipment was initially developed for bioterrorism assays following the anthrax attacks in the USA soon after the September 11 attacks in 2001. In this system, RNA extraction and real-time PCR is prepared.…”

Section: -177mentioning

confidence: 99%