Quantitative detection of PML-RARalpha fusion transcript by real-time PCR with a single primer pair.
Quantitative detection of minimal residual disease has prognostic value for some leukemias. Acute promyelocytic leukemia (APL) is characterized by the specific PML-RARalpha fusion gene from t(15;17). Added to three PML-RARalpha isoforms, alternative spliced forms of PML exons give rise to multiple i...
| Publicado en: | Journal of Clinical Laboratory Analysis Vol. 23; no. 4; pp. 223 - 231 |
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| Autores principales: | , , , , , , , , , , , , , , , |
| Formato: | research Journal Article |
| Publicado: |
Wiley-Blackwell
2009
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=105394738&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 105394738 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 08878013 OCK jtl: Journal of Clinical Laboratory Analysis issn: 08878013 maglogo: Y pubinfo: dt: 2009 vid: 23 iid: 4 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 105394738 105394738 NLM19623654 2010357368 10.1002/jcla.20306 NLM19623654 105394738 ppf: 223 ppct: 8 formats: tig: atl: Quantitative detection of PML-RARalpha fusion transcript by real-time PCR with a single primer pair. aug: au: Takenokuchi M Nakamachi Y Yoneda K Joo K Kawano S Tatsumi E Saigo K Kumagai S Takenokuchi, Mariko Nakamachi, Yuji Yoneda, Keiko Joo, Kana Kawano, Seiji Tatsumi, Eiji Saigo, Katsuyasu Kumagai, Shunichi affil: Faculty of Pharmacological Science, Himeji Dokkyo University, Himeji, Hyogo 670-8524, Japan sug: subj: Leukemia, Myeloid, Acute Proteins Receptors, Cell Surface Reverse Transcriptase Polymerase Chain Reaction Methods Bone Marrow Metabolism Bone Marrow Pathology Cells DNA Probes Leukemia, Myeloid, Acute Metabolism Prognosis Reproducibility of Results RNA Analysis Human ab: Quantitative detection of minimal residual disease has prognostic value for some leukemias. Acute promyelocytic leukemia (APL) is characterized by the specific PML-RARalpha fusion gene from t(15;17). Added to three PML-RARalpha isoforms, alternative spliced forms of PML exons give rise to multiple isoforms even within a single patient. To date, multiple primer pairs for the detection of the various PML-RARalpha transcripts have been designed, potentially generating some nonspecific amplification products. Here, we established a real-time quantitative PCR (RQ-PCR) strategy with a single primer pair using LightCycler (sp-RQ-PCR), which could simultaneously detect three isoforms with equal specificity and sensitivity as well as alternative spliced forms. Results obtained with sp-RQ-PCR for 39 samples from 15 APL patients and 31 non-APL samples were compared with those with TaqMan assay with three primer pairs. In two of the APL samples, PML-RARalpha was detected in the TM, but not in the sp-RQ-PCR or nested PCR. Furthermore, the sp-RQ-PCR showed no positive results for the 31 non-APL samples, whereas the TM identified 13% (4/31) as positive. Electrophoresis detected some artifacts in the TM, which do not correspond to PML-RARalpha. We conclude that our sp-RQ-PCR is specific enough to identify various forms of PML-RARalpha and yields no false-positive results. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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