Quantification of Bcr-Abl transcripts in chronic myelogenous leukemia (CML) using standardized, internally controlled, competitive differential PCR (CD-PCR).
The quantification of Bcr-Abl transcript numbers in chronic myelogenous leukemia (CML) patients described here uses simultaneous competitive PCR amplification of the target gene (Bcr-Abl) and a reference gene (porphobilinogen deaminase; Pbgd) together with a single composite competitor molecule for both targets based on heterologous sequences. Using this technique, Bcr-Abl transcript numbers could be reproducibly determined even in clinical samples known to harbour poor quality RNA.
Nagel
S
S
Abteilung für Innere Medizin m.S. Hämatologie und Onkologie, Virchow Klinikum, Medizinische Fakultät der Humboldt Universität zu Berlin, Germany.
Schmidt
M
M
Thiede
C
C
Huhn
D
D
Neubauer
A
A
D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D016313:Genes, abl / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D011163:Hydroxymethylbilane Synthase / Q000235:genetics; D007372:Interferons / Q000627:therapeutic use; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014158:Transcription, Genetic / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D007372:Interferons; D011163:Hydroxymethylbilane Synthase