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This protect aims to evaluate alternative strategies to screen for cervical cancerincluding liquid-based cytology (LBC) with supplementary reflex testing for humanpapillomavirus (HPV), p16INK4a, HPV-L1 capsid protein, and miR-205 for use aspossible diagnostic and prognostic markers in women with abnormal findings detectedthrough the organized cervical cancer screening program.This split-sample study enrolled 137 women with atypical Pap smear findings tocompare the effectiveness of conventional cytology (CC) with LBC cytology forcervical cancer screening. Sensitivity to detect cervical intraepithelial neoplasia (CIN2+) was 47% for CC compared with 66% for LBC. These results from these twosampling techniques agreed with histological findings in 37% and 53% of cases,respectively, a statistically significant difference. The significant advantage ofimproved sensitivity combined with the ability to carry out reflex testing for diagnosticand prognostic markers such as HPV DNA or p16 INK4a without repeated samplingfavors LBC for a possible future role as a screening technique.In cases of low-grade cytological abnormalities combining LBC with HPV triage(LBC+HPV triage) may improve detection of CIN compared with CC. Our study foundthat CC and LBC+HPV triage produced similar detection rates of CIN2+. Whencomparing CIN detection rates the adjusted OR for CIN2+ was 0.87 (95% CI: 0.60-1.26) and for CIN3+ 1.00 (95% CI: 0.64-1.58). Consequently, the use of LBC+HPVtriage offered no advantage over conventional cytology regarding sensitivity andspecificity or positive and negative predictive value for detection of histologicallyconfirmed CIN2+. Nevertheless, LBC+HPV triage may potentially reduce unnecessarymedical procedures and testing among HR-HPV-negative women with findings ofabnormal cytology.We analyzed 118 patient samples of dysplastic cells using immunocytochemicalstaining to assess the expression of p16INK4a. Expression levels of p16INK4a, correlatedwith CIN grade, showing stronger reactivity in higher grade lesions. We found astronger correlation between severity of cytological abnormality and p16INK4a,immunoreactivity when the diagnosis was simultaneously confirmed by routinecytology (p<0.001, χ2 exact test for trend). This staining technique may be able to serveas a complementary prognostic marker when used as a reflex test to identify women atrisk of cervical cancer.We also used immunocytochemistry to detect L1 capsid protein in HR-HPV-positivewomen who displayed minor cytological abnormalities. Progression to CIN2+ occurredin 2 of 13 L1-positive women (15.0%) infected with HPV16, compared with 4 L1-positive women infected with other HR-HPV types. Among all L1-positive womenwith CIN2+, 35.7% were infected with both HR and low-risk (LR) HPV types, 25.0%with HR-HPV types only and 13.3% with HPV16. Loss of previously positive L1expression may serve as a prognostic marker for preinvasive cervical lesions.We carried out reverse transcription quantitative PCR (RT-qPCR) on our LBC samplesto assess miR-205 expression and correlate these results with histology. Regardless ofwhether findings were based on histology or cytology, our results showed no significantchanges in miR-205 expression relating to different stages of disease progression. Itmay be that the increased expression of miR-205 is not stage-specific, but couldrepresent a continuous process throughout disease progression, although we cannotconclude this with certainty.Cervical cancer could become one of our most preventablecancers by effectively combining vaccination programs against HR-HPV withimproved detection of precursors by integrating molecular markers into screeningprograms

A comparison of liquid-based cytology and Pap smear as a screening method for cervical cancer