Method for discrimination of metaplasias from neoplastic or preneoplastic lesions
AssigneeVentana Medical Systems, Inc.
InventorsRuediger Ridder, Magnus Von Knebel Doeberitz, Peter Martin
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Abstract
The present invention relates to a method for discrimination of p16INK4a overexpressing metaplasias from neoplastic or preneoplastic p16INK4a overexpressing lesions by determination of the level of high risk HPV encoded gene-products such as e.g. HPV E2 and/or HPV E7 molecules in biological samples in the course of cytological testing procedures. The method thus enables for reduction of false positive results in the p16INK4a based detection of anogenital lesions in cytological testing procedures.
Claims20
A method for screening a uterine cervical cytology sample, comprising:
- (a) obtaining at least one slide having the uterine cervical cytology sample deposited thereon;
- (b) staining the uterine cytology sample with a labeled oligonucleotide probe that specifically binds p16INK4a mRNA and is complementary to p16INK4a mRNA in the sample by in situ hybridization, wherein the label of the oligonucleotide probe that is complementary to the p16INK4a mRNA generates a first reporter signal;
- (c) staining the uterine cervical cytology sample with a labelled oligonucleotide probe that specifically binds a high risk HPV (hr-HPV) gene-product and is complementary to a mRNA of the high risk HPV (hr-HPV) gene-product by in situ hybridization, wherein said high risk HPV gene-product is encoded by a gene selected from the group consisting of HPV E7 gene, HPV E2 gene, HPV E6 gene, HPV L1 gene, and HPV L2 gene, wherein the label of the oligonucleotide probe that is complementary to the mRNA of the hr-HPV gene product generates a second reporter signal;
- and
- (d) detecting in the uterine cytology sample:
- (d1) the simultaneous-presence of cells stained with the label generating the first reporter signal and cells stained with the label generating the second reporter signal, or
- (d2) the presence of cells stained with the label generating the first reporter signal and the absence of cells stained with the label generating the second reporter signal.
A method of preparing a uterine cervical cytology sample for microscopic evaluation, said method comprising:
- (a) obtaining a slide having the uterine cervical cytology sample deposited thereon;
- (b) staining the uterine cervical cytology sample on the slide by in situ hybridization for both:
- (b1) p16INK4a mRNA, wherein the stain for p16INK4a mRNA is an agent that specifically binds p16INK4a mRNA and gene rates a first reporter signal;
- and
- (b2) a high risk HPV (hrHPV) mRNA encoded by a gene selected from the group consisting of HPV E7 gene, HPV E2 gene, HPV E6 gene, HPV L1 gene, and HPV L2 gene, wherein the stain for the hrHPV is an agent that specifically binds the high risk HPV mRNA and generates a second reporter signal, wherein the first reporter signal is distinguishable from the second reporter signal.
A method of preparing a uterine cervical cytology sample for microscopic evaluation, said method comprising:
- (a) obtaining at least two slides having the uterine cervical cytology sample deposited thereon;
- (b) staining the uterine cervical sample on at least one of the slides by in situ hybridization for p16INK4a mRNA with an agent that specifically binds p16INK4a mRNA and generates a first reporter signal;
- and
- (c) staining the uterine cervical sample on at least one of the slides by in situ hybridization for a high risk HPV (hrHPV) mRNA with an agent that specifically binds the high risk HPV mRNA and generates a reporter signal, wherein the hrHPV is encoded by a gene selected from the group consisting of HPV E7 gene, HPV E2 gene, HPV E6 gene, HPV L1 gene, and HPV L2 gene, wherein the stain for the hrHPV is an agent that generates a second reporter signal, wherein the first reporter signal is distinguishable from the second reporter signal.
Dependencies are inferred from the claim text.