Diagnostic Performance of HPV E6/E7 mRNA and HPV DNA Assays for the Detection and Screening of Oncogenic Human Papillomavirus Infection among Woman with Cervical Lesions in China.

Hye-Young Wang,Geehyuk Kim,Hyeyoung Lee,Sunyoung Park,Kwang Hwa Park,Sunghyun Kim,Lin Han,Yingxue Li,Dongsup Lee,Ren Yubo

doi:10.7314/apjcp.2015.16.17.7633

Abstract

Human papillomavirus (HPV) is the most common sexually transmitted infection worldwide and it is responsible for most cases of cervical uterine cancer. Although HPV infections of the cervix do not always progress to cancer, 90% of cervical cancer cases have been found to be associated with high risk HPV (HR- HPV) infection. HPV DNA testing is widely used, along with Papanicolaou (Pap) testing, to screen for cervical abnormalities. However, there are no data on the prevalence of genotype-specific HPV infections assessed by measuring HPV E6/E7 mRNA in women representative of the Chinese population across a broad age range. In the present study, we compared the results with the CervicGen HPV RT-qDx assay, which detects 16 HR-HPV genotypes (Alpha-9: HPV 16, 31, 33, 35, 52, and 58; Alpha-7: HPV 18, 39, 45, 51, 59, and 68; and Alpha-5, 6: HPV 53, 56, 66, and 69), and the REBA HPV-ID assay, which detects 32 HPV genotypes based on the reverse blot hybridization assay (REBA) for the detection of oncogenic HPV infection according to cytological diagnosis. We also investigated the prevalence and genotype distribution of HPV infection with a total of 324 liquid-based cytology samples collected in western Shandong province, East China. The overall HPV prevalences determined by HPV DNA and HPV E6/E7 mRNA assays in this study were 79.9% (259/324) and 55.6% (180/324), respectively. Although the positivity of HPV E6/E7 mRNA expression was significantly lower than HPV DNA positivity, the HPV E6/E7 mRNA assay showed greater specificity than the HPV DNA assay (88.6% vs. 48.1%) in normal cytology samples. The prevalence of Alpha-9 (HPV 16, 31, 33, 35, 52, and 58) HPV infection among these women accounted for up to 80.3% and 76.1% of the high-grade lesions detected in the HPV mRNA and DNA tests, respectively. The HR-HPV genotype distribution, based on HPV DNA and E6/E7 mRNA expression by age group in patients with cytologically confirmed lesions, was highest in women aged 40 to 49 years (35.9% for cytologically confirmed cases, Pearson correlation r value=0.993, p<0.001) for high-grade lesions. Among the oncogenic HR-HPV genotypes for all age groups, there was little difference in the distribution of HPV genotypes between the HPV DNA (HPV -16, 53, 18, 58, and 33) and HPV E6/E7 mRNA (HPV -16, 53, 33, 58, and 18) assays. HPV 16 was the most common HPV genotype among women with high- grade lesions. Our results suggest that the HPV E6/E7 mRNA assay can be a sensitive and specific tool for the screening and investigation of cervical cancer. Furthermore, it may provide useful information regarding the necessity for early cervical cancer screenings and the development of additional effective HPV vaccines, such as one for HPV 53 and 58. Additionally, gaining knowledge of HPV distribution may also inform us about ecological changes in HPV after the vaccination.

Highlights

Human uterine cervical cancer is the third most common cancer among women worldwide (Jemal et al., 2011) and a major cause of morbidity and mortality
Materials and Methods: In the present study, we compared the results with the CervicGen Human papillomavirus (HPV) reverse transcriptase (RT)-qDx assay, which detects 16 HR-HPV genotypes (Alpha-9: HPV 16, 31, 33, 35, 52, and 58; Alpha-7: HPV 18, 39, 45, 51, 59, and 68; and Alpha-5, 6: HPV 53, 56, 66, and 69), and the REBA HPV-ID assay, which detects 32 HPV genotypes based on the reverse blot hybridization assay (REBA) for the detection of oncogenic HPV infection according to cytological diagnosis
In the present study we used the CervicGen HPV RT-qDx assay (Optipharm, Osong, Republic of Korea), which targets HPV E6/E7 mRNA of 16 oncogenic HR HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, and 69) to investigate the prevalence and genotype distribution in relation to age in Chinese women, cytology of oncogenic HPV infection compared with the REBA HPV-ID which is based on the reverse blot hybridization assay (REBA)

Summary

Introduction

Human uterine cervical cancer is the third most common cancer among women worldwide (Jemal et al., 2011) and a major cause of morbidity and mortality. As a result, testing for oncogenic HPV infection in cervical lesions could serve as an accurate means of identifying women who are at risk for developing cervical cancer. As a supplement to the Pap smear, the HPV molecular diagnosis test is effective in diagnosing HR-HPV groups in cervical cancer. For uterine cervical cancer diagnosis, most molecular diagnostics are based on an HPV DNA test with Pap smear. It has been reported that the HPV E6/E7 mRNA test has greater specificity for detecting cancer development than do L1 gene-based DNA tests (Sveinung et al, 2010; Andersson et al, 2011), there are no data on the distribution of typespecific HPV infections among HPV mRNA detected in women representative of the Chinese population covering a broad age range. In the present study we used the CervicGen HPV RT-qDx assay (Optipharm, Osong, Republic of Korea), which targets HPV E6/E7 mRNA of 16 oncogenic HR HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, and 69) to investigate the prevalence and genotype distribution in relation to age in Chinese women, cytology of oncogenic HPV infection compared with the REBA HPV-ID which is based on the reverse blot hybridization assay (REBA)

Materials and Methods

Results

Discussion