HPV E6, E7 mRNA assay and methods of use thereof
First Claim
1. An assay for determining malignant transformation of cervical cells, the assay comprising the steps of:
- (a) obtaining cervical cells from an individual;
(b) preparing the cervical cells as a liquid-based cytology (LBC) specimen;
(c) hybridizing the LBC cytology specimen using oligonucleotides specific for HPV E6, E7 mRNA;
(d) quantitating expression of HPV E6, E7 mRNA in the cervical cells; and
(e) identifying cervical cells that express 200 or more copies of HPV E6, E7 mRNA per cell,wherein malignant transformation of the cervical cells is indicated by expression of 200 or more copies of HPV E6, E7 mRNA per cell and wherein the percentage of cells that express 200 or more copies of HPV E6, E7 mRNA per cell is quantitated.
2 Assignments
0 Petitions
Accused Products
Abstract
Provided is an HPV E6, E7 mRNA assay, referenced herein as the “In Cell HPV Assay,” that is capable of sensitive and specific detection of normal cervical cells undergoing malignant transformation as well as abnormal cervical cells with pre-malignant or malignant lesions. The In Cell HPV Assay identifies HPV E6, E7 mRNA via in situ hybridization with oligonucleotides specific for HPV E6, E7 mRNA and quantitates the HPV E6, E7 mRNA via flow cytometry. The In Cell HPV Assay can be carried out in less than three hours directly from liquid-based cervical (“LBC”) cytology specimens. The In Cell HPV Assay provides an efficient and highly sensitive alternative to the Pap smear for determining abnormal cervical cytology.
-
Citations
9 Claims
-
1. An assay for determining malignant transformation of cervical cells, the assay comprising the steps of:
-
(a) obtaining cervical cells from an individual; (b) preparing the cervical cells as a liquid-based cytology (LBC) specimen; (c) hybridizing the LBC cytology specimen using oligonucleotides specific for HPV E6, E7 mRNA; (d) quantitating expression of HPV E6, E7 mRNA in the cervical cells; and (e) identifying cervical cells that express 200 or more copies of HPV E6, E7 mRNA per cell, wherein malignant transformation of the cervical cells is indicated by expression of 200 or more copies of HPV E6, E7 mRNA per cell and wherein the percentage of cells that express 200 or more copies of HPV E6, E7 mRNA per cell is quantitated. - View Dependent Claims (2, 3)
-
-
4. A method of screening a patient for malignant transformation of cervical cells, the method comprising:
-
(a) obtaining cervical cells from the patient; (b) quantitating expression of HPV E6, E7 mRNA in the cervical cells; and (c) identifying cervical cells that express 200 or more copies of HPV E6, E7 mRNA per cell, wherein malignant transformation of cervical cells in the patient is indicated by expression of 200 or more copies of HPV E6, E7 mRNA per cell and wherein the percentage of cells that express 200 or more copies of HPV E6, E7 mRNA per cell is quantitated. - View Dependent Claims (5, 6, 7)
-
-
8. An assay for identifying one or more malignantly transformed cervical cell types comprising the steps of:
-
(a) obtaining cervical cells from an individual; (b) preparing the cervical cells as a liquid-based cytology (LBC) specimen; (c) staining the cervical cells with a combination of CD16 and CAM 5.2; (d) hybridizing the LBC specimen using oligonucleotides specific for HPV E6, E7 mRNA, (e) quantitating expression of HPV E6, E7 mRNA in the cervical cells; (f) identifying cells that express 200 or more copies of HPV E6, E7 mRNA per cell, wherein neutrophils are identified by CD16.sup.+ staining, columnar endocervical cells are identified by CD16.sup.−
, CAM 5.2.sup.+ staining, and squamous ectocervical cells are identified by CD16.sup.−
, CAM 5.2.sup.−
staining, and malignant transformation of one or more of the cervical cell types is indicated by expression of 200 or more copies of HPV E6, E7 mRNA per cell and wherein the percentage of cells that express 200 or more copies of HPV E6, E7 mRNA per cell is quantitated.- View Dependent Claims (9)
-
Specification