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Titolo:
Pre-clinical validation of a novel, highly sensitive assay to detect PML-RAR alpha mRNA using real-time reverse-transcription polymerase chain reaction
Autore:
Slack, JL; Bi, WL; Livak, KJ; Beaubier, N; Yu, M; Clark, M; Kim, SH; Gallagher, RE; Willman, CL;
Indirizzi:
Roswell Pk Canc Inst, Dept Med, Buffalo, NY 14263 USA Roswell Pk Canc Inst Buffalo NY USA 14263 Dept Med, Buffalo, NY 14263 USA Appl Biosyst Inc, Foster City, CA 94404 USA Appl Biosyst Inc Foster City CA USA 94404 Inc, Foster City, CA 94404 USA Univ New Mexico, Canc Res Facil, Dept Pathol, Albuquerque, NM 87131 USA Univ New Mexico Albuquerque NM USA 87131 athol, Albuquerque, NM 87131 USA Montefiore Med Ctr, Dept Oncol, Bronx, NY 10467 USA Montefiore Med Ctr Bronx NY USA 10467 tr, Dept Oncol, Bronx, NY 10467 USA Albert Einstein Canc Ctr, Bronx, NY USA Albert Einstein Canc Ctr Bronx NYUSA t Einstein Canc Ctr, Bronx, NY USA
Titolo Testata:
JOURNAL OF MOLECULAR DIAGNOSTICS
fascicolo: 4, volume: 3, anno: 2001,
pagine: 141 - 149
SICI:
1525-1578(200111)3:4<141:PVOANH>2.0.ZU;2-J
Fonte:
ISI
Lingua:
ENG
Soggetto:
ACUTE PROMYELOCYTIC LEUKEMIA; MINIMAL RESIDUAL DISEASE; QUANTITATIVE RT-PCR; LONG-TERM REMISSION; FUSION TRANSCRIPTS; RETINOIC ACID; CELL-LINE; PERSISTENCE; RNA; TRANSLOCATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Citazioni:
28
Recensione:
Indirizzi per estratti:
Indirizzo: Slack, JL Roswell Pk Canc Inst, Dept Med, Elm & Carlton St, Buffalo, NY 14263 USA Roswell Pk Canc Inst Elm & Carlton St Buffalo NY USA 14263 3 USA
Citazione:
J.L. Slack et al., "Pre-clinical validation of a novel, highly sensitive assay to detect PML-RAR alpha mRNA using real-time reverse-transcription polymerase chain reaction", J MOL DIAGN, 3(4), 2001, pp. 141-149

Abstract

We have developed a sensitive and quantitative reverse-transcription polymerase chain reaction (RT-PCR) assay for detection of PML-RA-R alpha, the fusion oncogene present as a specific marker in > 99% of cases of acute promyelocytic leukemia (APL). The assay is linear over at least 5 orders of magnitude of input DNA or RNA, and detects as few as 4 copies of PML-RAR alpha plasmid DNA. PML-RA-R alpha transcripts could be detected in mixtures containing 2 to 5 pg of RNA from fusion-containing cells in a background of 1 mug of RNA from PML-RAR alpha -negative cells. Using 1.0 to 2.5 mug of input RNA, the sensitivity of the assay was between 10(-5) and 10(-6). Furthermore, determination of GAPDH copy number in each reaction allowed an accurate assessment of sample-to-sample variation In RNA quality and reaction efficiency, with consequent definition of a detection limit for each sample assayed. Using an internal calibrator, assay precision was high, with coefficients of variation between 10 and 20%. An interlaboratory study using coded samples demonstrated excellent reproducibility and high concordance between Laboratories. This assay will be used to test the hypothesis that sensitive and quantitative measurement of leukemic burden, during or after therapy ofAPL, can stratify patients into discrete risk groups, and thereby serve asa basis for risk-adapted therapy in APL.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/04/20 alle ore 15:16:35