Co-detection of chimeric bcr/abl (target) and β-actin (control) messenger RNA in individual CFU-GM colonies derived from CML patients using the polymerase chain reaction Journal Article

  

Authors:	Wisniewski, D.; Strife, A.; Clarkson, B.
Article Title:	Co-detection of chimeric bcr/abl (target) and β-actin (control) messenger RNA in individual CFU-GM colonies derived from CML patients using the polymerase chain reaction
Abstract:	In order to quantitate the magnitude of the normal and Philadelphia (Ph′) chromosome-positive(+) progenitor cells for various research and clinical settings/studies, we have applied the highly sensitive polymerase chain reaction (PCR) for examining the cells contained in individual hematopoietic colonies for chimeric bcr/abl mRNA, a specific molecular marker for chronic myelogenous leukemia (CML). Thus, individual 14-day CFU-GM colonies, obtained by growth of bone marrow cells from CML patients were removed from methylcellulose cultures and total RNA from each colony was isolated. First-strand complementary DNAs (cDNA) corresponding to all mRNAs in the sample were obtained by using random hexamers in a reverse transcription (RT) reaction. cDNA then served as the substrate in the PCR. To ensure the integrity of the RNA extracted from each colony, β-actin and bcr/abl cDNA sequences were amplified in the same reaction vessel. Using this method, we have examined the colonies grown from threee CML patients and found that 5 out of 5, 9 out of 9 and 8 out of 9 colonies contained a bcr/abl transcript. This method is simple, highly sensitive and should facilitate studies comparing the expression of various oncogenes in normal and leukemic hematopoietic progenitor cells. © 1991.
Keywords:	controlled study; human cell; proto-oncogene proteins; polymerase chain reaction; gene expression; tumor markers, biological; transcription factor; chronic myeloid leukemia; chimera; oncogene; messenger rna; rna, messenger; bone marrow cell; actins; macrophages; complementary dna; in vitro; colony forming unit gm; chronic myelogenous leukemia; beta actin; philadelphia chromosome; proto-oncogene proteins c-abl; granulocytes; gene rearrangement, b-lymphocyte; leukemia, myeloid, chronic; blotting, southern; oligonucleotide probes; rna directed dna polymerase; human; priority journal; article; cfu-gm; electrophoresis, agar gel; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; bcr/abl mrna
Journal Title:	Leukemia Research
Volume:	15
Issue:	9
ISSN:	0145-2126
Publisher:	Elsevier Ltd
Date Published:	1991-01-01
Start Page:	867
End Page:	874
Language:	English
DOI:	10.1016/0145-2126(91)90471-5
PUBMED:	1921462
PROVIDER:	scopus
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-0025773396&doi=10.1016%2f0145-2126%2891%2990471-5&partnerID=40&md5=f24e7f79ef57187dd198b81794a550c7
Notes:	Source: Scopus

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