Identification of neoplasms by detection of genetic insertions and deletions
View Patent ↗The detection of insertions and/or deletions in reiterated nucleotide sequences in tissues provides an identification of neoplastic changes that are associated with malignancy. The mutations are preferably detected by PCR based amplification of target sequences using selected primers, followed by standard analytic procedures. The detection of these mutations is useful as a diagnostic tool for cancer development and has direct application for cancer prognosis.
1. A diagnostic kit for providing a prognosis for at least one of colorectal, stomach, and pancreatic tumor cells:
at least one set of arbitrary primer pairs complementary to a predetermined portion of DNA from said tumor cells, wherein the predetermined portion of DNA comprises one or more mutations, wherein the mutations are at least one of deletions and insertions of one or a few nucleotides in reiterated sequences or microsatellite sequences, wherein the reiterated sequences are characterized as having multiple adjacent occurrences of the same nucleotide, dinucleotide pair, or nucleotide triplet, and wherein the microsatellite sequences are characterized as having 1 to 6 base pair tandem repeats; and
reagents to carry out PCR amplification;
wherein the at least one of colorectal, stomach, and pancreatic tumor cells have a prognosis of at least one of the following:
(i) being less invasive than tumors without said mutations;
(ii) having a lower rate of recurrence than tumors without said mutations; and
(iii) being less likely to have metastasis than tumors without said mutations.
2. The kit of claim 1 , wherein said tumor cells are colorectal tumor cells.
3. The kit of claim 1 wherein said tumor cells are stomach tumor cells.
4. The kit of claim 1 , wherein said tumor cells are pancreatic tumor cells.
5. A diagnostic kit for providing a prognosis for at least one of colorectal, stomach, and pancreatic tumor cells, comprising:
at least one pair of ligase chain reaction primers complementary to a predetermined region of DNA from said tumor cells, wherein the predetermined portion of DNA comprises one or more mutations, wherein the mutations are at least one of deletions and insertions of one or a few nucleotides in reiterated sequences or microsatellite sequences, wherein the reiterated sequences are characterized as having multiple adjacent occurrences of the same nucleotide, dinucleotide pair, or nucleotide triplet, and wherein the microsatellite sequences are characterized as having 1 to 6 base pair tandem repeats, amplification occurring only if one or more insertions or deletions have occurred in said reiterated sequences contained within said region; and
reagents for performing ligase chain reaction using said primers;
wherein the at least one of colorectal, stomach, and pancreatic tumor cells have a prognosis of at least one of the following:
(i) being less invasive than tumors without said mutations;
(ii) having a lower rate of recurrence than tumors without said mutations; and
(iii) being less likely to have metastasis than tumors without said mutations.
6. The kit of claim 5 , further comprising a second pair of primers adapted to amplify said region only if a different, second number of insertions or deletions have occurred in said reiterated sequence.
7. The kit of claim 5 , further comprising a third pair of primers complementary to said region, amplification occurring only if a third number of insertions or deletions has occurred in said reiterated sequence, wherein said third number is different from said first and second numbers.
8. The kit of claim 5 , wherein said tumor cells are colorectal tumor cells.
9. The kit of claim 5 , wherein said tumor cells are stomach tumor cells.
10. The kit of claim 5 , wherein said tumor cells are pancreatic tumor cells.
11. The kit of claim 1 , further comprising at least one of control DNA samples, a polyacrylamide gel slab, and electrophoresis buffer.
12. The kit of claim 5 , further comprising at least one of control DNA samples, a polyacrylamide gel slab, and electrophoresis buffer.
13. The kit of claim 1 , wherein the at least one of colorectal, stomach, and pancreatic tumor cells have a prognosis of being less invasive than tumors without said mutations.
14. The kit of claim 5 , wherein the at least one of colorectal, stomach, and pancreatic tumor cells have a prognosis of being less invasive than tumors without said mutations.