Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1988-03-31
1991-03-12
Wax, Robert A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 27, 935 21, 935 77, 935 78, C12Q 168, C07H 1512, C12N 1500
Patent
active
049992902
ABSTRACT:
The present invention involves a method for detecting the unique aberrant gene transcripts of a targeted cellular genomic abnormality in a tissue sample. This method comprises a series of steps. Initially, total cellular RNA or m-RNA is preferred from the tissue sample to be analyzed for the presence of a genomic abnormality. The total cellular RNA or m-RNA is then mixed with at least one synthetic DNA oligonucleotide complementary to the unique RNA sequence of the targeted cellular genomic abnormality being detected. The mixing is under conditions facilitating formation of double stranded DNA-RNA heteroduplexes when a strand of synthetic DNA oligonucleotide is complementary to an RNA strand obtained from the tissue sample. The conditions are those such as time, salt concentration, temperature and pH 10. The synthetic DNA oligonucleotide is preferably about several hundred nucleotides in length, more preferably about 60 to about 150 nucleotides in length.
In a most preferred embodiment, the method of the present invention may be applied to the detection of residual cells of chronic myelogenous leukemia with its characteristic Philadelphia chromosome (Ph.sup.1). For this situation, the first primer is primer bcr Ex III(+) or bcr Ex II (+), most preferably a mixture of primer bcr Ex III (+) and bcr Ex II (+). In certain instances the second primer may also be a mixture of two different primers, particularly where there is a variable breakpoint in the second chromosomal member of the translocation characteristic of a neoplasm. Where residual cells of chronic myelogenous leukemia are being detected, the second primer is most preferably primer abl (-).
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Fleisher M.
The Board of Regents The University of Texas System
Wax Robert A.
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