Raman Spectroscopy of Liquid-Based Cervical Smear Samples as a Triage to Stratify Women Who Are HPV-Positive on Screening.

John J. O’Leary,Fiona M. Lyng,Christine White,Stephen Reynolds,Cara M. Martin,Tom D’Arcy,Damien Traynor

doi:10.3390/cancers13092008

Abstract

Simple SummaryPersistent high-risk human papillomavirus (HPV) infection can lead to cervical precancer and cancer. Recently, HPV testing has been introduced for primary cervical screening, but the HPV DNA test cannot distinguish between transient and persistent HPV infection. Thus, there is an unmet clinical need to develop a new test to identify women with a high-risk persistent HPV infection. Raman spectra were recorded from cervical smear samples (n = 60) and, on the basis of HPV DNA and HPV mRNA test results, a classifier was developed to identify persistent HPV infection. A further blinded independent test set (n = 14) was used to validate the model, and sensitivity of 90% and specificity of 100% were achieved. Improved triage would allow women with a high-risk persistent HPV infection to be referred for immediate treatment, while women with a low-risk transient infection could avoid overtreatment.The role of persistent high-risk human papillomavirus (HPV) infection in the development of cervical precancer and cancer is now well accepted, and HPV testing has recently been introduced for primary cervical screening. However, the low specificity of HPV DNA testing can result in large numbers of women with an HPV-positive result, and additional triage approaches are needed to avoid over-referral to colposcopy and overtreatment. The aim of this study was to assess Raman spectroscopy as a potential triage test to discriminate between transient and persistent HPV infection. HPV DNA status and mRNA status were confirmed in ThinPrep® cervical samples (n = 60) using the Cobas 4800 and APTIMA HPV test, respectively. Raman spectra were recorded from single-cell nuclei and subjected to partial least squares discriminant analysis (PLSDA). In addition, the PLSDA classification model was validated using a blinded independent test set (n = 14). Sensitivity of 85% and specificity of 92% were achieved for the classification of transient and persistent HPV infection, and this increased to 90% sensitivity and 100% specificity when mean sample spectra were used instead of individual cellular spectra. This study showed that Raman spectroscopy has potential as a triage test for HPV-positive women to identify persistent HPV infection.

Highlights

Cervical cancer ranks fourth for both incidence and mortality with an estimated570,000 cases and 311,000 deaths worldwide in 2018 [1]
The scatter plot shows that latent variable LV1 mainly contributed to the differentiation of samples with non-transcriptionally active and transcriptionally active human papillomavirus (HPV) infection (Figure 2b)
The loadings from LV1 are shown in Figure 2c and show that discrimination is based around Raman peaks at 482, 782, 826, 852, 937, 1123, 1152, 1334, 1380, 1238, 1450, 1485, 1580, 1642, and 1670 cm−1

Summary

Introduction

Cervical cancer ranks fourth for both incidence and mortality with an estimated570,000 cases and 311,000 deaths worldwide in 2018 [1]. Cervical cancer ranks fourth for both incidence and mortality with an estimated. Health Organization’s call to action to eliminate cervical cancer, improved screening and early detection are required [2,3]. Persistent infection with high-risk human papillomavirus (HPV) is accepted as the major cause of the development of cervical precancer and cancer [4]. Over 100 different types of HPV have been identified, and 14 are considered as high-risk HPV types (hrHPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) [5]. Persistent hrHPV infection, followed by integration of the HPV genome into the host chromosomes, results in production of E6/E7 messenger RNA (mRNA) transcripts and

Objectives

Methods

Results

Discussion

Conclusion