Methods for detecting human papilloma virus-associated cancers
The present invention provides probes and methods of use thereof in the diagnosis and/or prognosis of certain types of cancers, particularly human papillomavirus (HPV)-associated cancers. The probes are designed for hybridization with genomic material in a manner indicative of one or more aberrations in the genetic material present in the test sample. The identified aberrations are biomarkers of HPV-associated cancer. The methods of the invention comprise contacting a sample to one or more probes, allowing any genetic material in the sample to hybridize to the genomic regions provided in the probes, analyzing the hybridizations, and analyzing the hybridizations to identify detected aberrations as biomarkers indicative of HPV-associated cancer progression.
1. A method of identifying an individual with one or more human chromosomal abnormalities, said method comprising:
(a) obtaining a sample from an individual, said sample comprising genetic material from cervical tissue and/or fluid thereof;
(b) hybridizing a probe set with the genetic material, wherein hybridizing comprises fluorescence in situ hybridization (FISH), and wherein the probe set consists of
(i) a first probe that hybridizes selectively to 3q26, said first probe comprising a probe preparation of seven different nick-translated clones corresponding to chromosome 3, clones 1-7 of Table 4,
(ii) a second probe that hybridizes selectively to 5p15, said second probe comprising a probe preparation of five different nick-translated clones corresponding to chromosome 5, clones 1-5 of Table 4,
(iii) a third probe that hybridizes selectively to 20q13, said third probe comprising a probe preparation of three different nick-translated clones corresponding to chromosome 20, clones 1-3 of Table 4, and
(iv) a fourth probe that hybridizes selectively to cen7, said fourth probe comprising a probe preparation of a nick-translated clone corresponding to chromosome 7, clone 1 of Table 4; and
(c) analyzing the hybridization pattern of the probe set to the genetic material to determine the presence or absence in the sample of a gain of 3q, a gain of 5p, a gain of 20q, and a gain of cen7.
2. The method of claim 1 , wherein the genetic material comprises chromosomes.
3. The method of claim 1 , wherein the identified individual is in a precancerous stage of an HPV-associated cancer.
4. The method of claim 1 , wherein said probe that hybridizes to 3q26 is capable of hybridizing to SEQ ID NO: 1 or complement thereof.
5. The method of claim 1 , wherein said probe that hybridizes to 5p15 is capable of hybridizing to SEQ ID NO: 2 or complement thereof.
6. The method of claim 1 , wherein said probe that hybridizes to 20q13 is capable of hybridizing to SEQ ID NO: 3 or complement thereof.
7. The method of claim 1 , wherein said probe that hybridizes to cen7 is capable of hybridizing to SEQ ID NO: 4 or complement thereof.
8. A method of identifying in a sample one or more chromosomal abnormalities, said method comprising:
(a) obtaining a sample from an individual, said sample comprising genetic material from cervical tissue and/or fluid thereof;
(b) hybridizing a probe set with the genetic material, wherein hybridizing comprises fluorescence in situ hybridization (FISH), and wherein the probe set consists of
(i) a first probe that hybridizes selectively to 3q26, said first probe comprising a probe preparation of seven different nick-translated clones corresponding to chromosome 3, clones 1-7 of Table 4,
(ii) a second probe that hybridizes selectively to 5p15, said second probe comprising a probe preparation of five different nick-translated clones corresponding to chromosome 5, clones 1-5 of Table 4,
(iii) a third probe that hybridizes selectively to 20q13, said third probe comprising a probe preparation of three different nick-translated clones corresponding to chromosome 20, clones 1-3 of Table 4, and
(iv) a fourth probe that hybridizes selectively to cen7, said fourth probe comprising a probe preparation of a nick-translated clone corresponding to chromosome 7, clone 1 of Table 4; and
(c) analyzing the hybridization pattern of the probe set to the genetic material to determine the presence or absence in the sample of a gain of 3q, a gain of 5p, a gain of 20q, and a gain of cen7.
9. The method of claim 1 , wherein each of said probes is labeled with a different color fluorochrome.