Detection of t(9;22) b2a2 fusion transcript by flow cytometry.
Introduction We assessed the feasibility of flow cytometry-fluorescent in situ hybridization technique in the detection of translocated mRNA in the cytoplasm of human peripheral blood nucleated cells. It is assumed that this assay can be applied as a diagnostic method in the detection of chromosomal...
| Published in: | International Journal of Laboratory Hematology Vol. 38; no. 4; pp. 403 - 412 |
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| Main Authors: | , , , , , , , , |
| Format: | pictorial research tables/charts Journal Article |
| Published: |
Wiley-Blackwell
Aug2016
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| Online Access: | View this record in EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=116790916&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 116790916 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 17515521 47EA jtl: International Journal of Laboratory Hematology issn: 17515521 maglogo: Y pubinfo: dt: Aug2016 vid: 38 iid: 4 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 116790916 116790916 116790916 10.1111/ijlh.12515 116790916 ppf: 403 ppct: 9 formats: tig: atl: Detection of t(9;22) b2a2 fusion transcript by flow cytometry. aug: au: Ranjbaran, R. Okhovat, M. A. Abbasi, M. Moezzi, L. Aboualizadeh, F. Amidzadeh, Z. Golafshan, H. A. Behzad‐Behbahani, A. Sharifzadeh, S. affil: Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz Iran sug: subj: Leukemia, Myeloid, Chronic Diagnosis Chromosome Aberrations Flow Cytometry In Situ Hybridization, Fluorescence Sensitivity and Specificity Human RNA, Messenger DNA Probes Reverse Transcriptase Polymerase Chain Reaction Microscopy Methods Cell Line, Tumor ab: Introduction We assessed the feasibility of flow cytometry-fluorescent in situ hybridization technique in the detection of translocated mRNA in the cytoplasm of human peripheral blood nucleated cells. It is assumed that this assay can be applied as a diagnostic method in the detection of chromosomal translocation which commonly occurs in hematologic malignancies. Methods KCL-22 cell line and white blood cells from 21 CML patients were recruited in the study. Cells were isolated and fixed. After permeabilization, cells were resuspended in hybridization buffer and probes were added to the mixture. Subsequently, cells were washed and analyzed on the flow cytometer instrument. The flow cytometry results were compared with qRT- PCR and fluorescent microscope outcomes. Results Using the current principle, 97 ± 2.1% of the KCL-22 cells were labeled with b2a2 mRNA-specific probes. In addition, seven patients were recognized positive for t(9;22) b2a2. The percentage of cells containing abovementioned translocation in these patients was varied from 3.26% to 97.8%. There was no false-positive result in negative controls (K562 with BCR- ABL1 b3a2, NB4, and Jurkat cell lines) along with blood samples of normal controls. All the results obtained by flow- FISH were confirmed by qRT- PCR and fluorescent microscope. Conclusion This strategy benefits from appropriate specificity, sensitivity, rapidity, and ability in the determination of malignant cell percentage. Therefore, it can improve traditional time-consuming and labor-intensive FISH method. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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