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 (-).
The invention is directed to a DNA probe set, the probe set comprising a first probe set and a second probe set, the first probe set being sufficient in length and substantially complementary to an entire breakpoint region of a first DNA and nucleotides breakpoint region but less than an entire chromosome such that the first probe set will hybridize to both sides of the breakpoint region regardless of whether the first DNA has been broken in the breakpoint region and either end fused to another DNA, and the second probe set being sufficient in length and substantially complementary to an entire breakpoint region of a second DNA and nucleotides on both sides of the breakpoint region but less than an entire chromosome such that the second probe set will hybridize to both sides of the breakpoint region regardless of whether the second DNA has been broken in the breakpoint region and either end fused to another DNA. Diagnostic kits utilizing the probe sets of the invention are also claimed.
The present invention provides novel anticode oligomers and methods of using them for controlling the growth of cancer cells expressing the bcl-2 gene.
A method of detecting reciprocal gene translocations by employing multiple probe sets which span the breakpoint regions and encompass regions on both sides of each breakpoint of the translocation on each chromosome involved in the translocation. Probe sets and diagnostic kits for the method are also disclosed.
The present invention provides novel anticode oligomers and methods of using them for controlling the growth of cancer cells expressing the bcl-2 gene.
