Method for the detection of malignant and premalignant stages of cervical cancer

• US 6,031,232 A
• Filed: 11/13/1995
• Issued: 02/29/2000
• Est. Priority Date: 11/13/1995
• Status: Expired due to Term

• Current Assignee: Agilent Technologies Australia (M) Pty., Ltd. (Agilent Technologies, Inc.)
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Rigas, Basil, Cohenford, Menashi A., Bhandare, Prashant S., Krishnan, Krishnaswamy, Cahn, Frederick R.
• Primary Examiner(s): Smith, Ruth S.
• Application Number:
  US08/558,130
• Time in Patent Office: 1,569 Days
• Field of Search: 128/664, 128/665, 250/339.01, 250/339.06, 250/341.1, 250/339.12, 250/339.09
• US Class Current: 250/339.09
• CPC Class Codes: G01J 2003/2866 Markers; Calibrating of scan
• CPC Class Codes: G01J 3/28 Investigating the spectrum using colour filters G01J3/51
• CPC Class Codes: G01J 3/453 by correlation of the amplitudes
• CPC Class Codes: G01N 2021/3595 using FTIR
• CPC Class Codes: G01N 21/35 using infrared light G01N21/39 takes precedence

Abstract

This invention discloses a method to identify premalignant and malignant stages of cervical cancer from an infrared (IR) spectrum of exfoliated cervical cells which are dried on an infrared transparent matrix and scanned at the frequency range from 3000-950 cm^-1. The identification of samples is based on establishing a calibration using a representative set of spectra of normal, dysplastic and malignant specimens. During the calibration process, multivariate techniques such as Principal Component Analysis (PCA) and/or Partial Least Squares (PLS) are used. PCA and PLS reduce the data based on maximum variations between the spectra, and generate clusters in a multidimensional space representing the different populations. The utilization of Mahalinobis distances, or linear regression (e.g., Principle Component Regression on the reduced data from PCA) form the basis for the discrimination. This method is simple to use and achieves statistically reliable distinction between the following groups of cervical smears: normal (individuals with no prior history of dysplasia), dysplasia and malignant samples. Lastly, this invention discloses a method to obtain the IR spectrum of individual cervical cells fixed on an infrared transparent matrix.

Claims

1. A method for the identification of a malignant or premalignant condition in an exfoliated cervical cell sample, said method comprising:

(a) drying said exfoliated cervical cell sample on an infrared transparent matrix to produce a dried cell sample;

(b) directing a beam of mid-infrared light at said dried cell sample, said beam of mid-infrared light having a frequency of from about 3000 to about 950 cm^-1 to produce absorption data for said dried cell sample; and

(c) comparing said absorption data for said dried cell sample with a calibration/reference set of infrared absorption data to determine whether variation in infrared absorption occurs in said dried cell sample, at at least one range of frequencies, due to the said variation being characteristic of said malignant or premalignant condition, said comparing utilizing a partial least squares or principal component analysis statistical method and said absorption data being underivatized and unsmoothed, whereby said identification of said malignant or premalignant condition is made.

2. A method in accordance with claim 1 wherein said calibration/reference set of infrared absorption data is from a representative set of normal, dysplastic and malignant cervical cells.

3. A method in accordance with claim 2, wherein said calibration/reference set is prepared from about 100 to about 1000 reference cell samples.

4. A method in accordance with claim 2, wherein said calibration/reference set of infrared absorption data is prepared from about 100 to about 500 reference cell samples.

5. A method in accordance with claim 1, wherein said comparing utilizes principal component regression which is carried out using principal component analysis.

6. A method in accordance with claim 1, wherein said infrared transparent matrix is a matrix prepared from a member selected from the group consisting of BaF₂, ZnS, polyethylene film, CsI, KCl, KBr, CaF₂, NaCl and ZnSe.

7. A method in accordance with claim 1, wherein prior to step (a) said exfoliated cervical cell sample is dispersed, thereby separating said cervical cells from nondiagnostic debris in said sample to provide a substantially uniform suspension of cells for drying.

8. A method in accordance with claim 7, wherein said exfoliated cervical cell sample is dispersed in a preservative solution.

9. A method in accordance with claim 1, wherein said comparing utilizes principal component analysis and is confined to the frequency region of about 1200 cm^-1 to about 1000 cm^-1.

10. A method in accordance with claim 1, wherein said comparing utilizes principal component analysis and is carried out by concurrent analysis of the frequency regions of about 1250 to 1000 cm^-1, about 1420 to 1330 cm^-1 and about 3000 to 2800 cm^-1.

11. A method in accordance with claim 1, wherein said beam of mid-infrared light is directed through an aperture of individual cell size and said absorption data for said dried cell sample is produced for single cells.

12. A method in accordance with claim 1, wherein prior to step (a) said exfoliated cervical cell sample is dispersed in a preservative solution, thereby separating said cervical cells from nondiagnostic debris in said sample to provide a substantially uniform suspension of cells for drying and wherein said beam of mid-infrared light is directed through an aperture of individual cell size and said absorption data for said dried cell sample is produced for single cells.