title: Performance of HPV E6/E7 mRNA RT-qPCR for screening and diagnosis of cervical cancer with ThinPrep Pap test samples.
abstract: Recent research has shown that oncogenic human papillomavirus (HPV) DNA, which is currently used in the screening and diagnosis of cervical cancer, can be detected not only in high-grade cervical lesions, but also in low-grade cervical lesions and normal tissues. For this reason, HPV tests targeting the E6 and E7 mRNA of five oncogenic HPV strains (HPV genotypes 16, 18, 31, 33, and 45), which are known to be responsible for the oncogenesis of cervical cancer, have been commercialized using a real-time nucleic acid sequence based amplification (NASBA) assay. Previous data has shown that the real-time NASBA assay has higher clinical specificity than HPV DNA testing (97.1% vs. 53.7%). However, the sensitivity of the real-time NASBA assay was lower than that of HPV DNA testing (41.1% vs. 100%). Despite the fact that there are more than 16 oncogenic HPV genotypes known to cause cervical cancer (HPV genotypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, and 69), the commercialized real-time NASBA kit was designed to detect only five genotypes (16, 18, 31, 33, and 45). Therefore, in the present study, CervicGen HPV RT-qDX (Optipharm), a commercial diagnostic kit targeting a HPV E6/E7 mRNA based on RT-qPCR assay was evaluated with RNA extracted from ThinPrep Pap samples, and the results were compared to real-time NASBA data. The sensitivity and specificity of the RT-qPCR assay were 91% and 98.6%, respectively, for the detection of cervical intraepithelial neoplasia CIN2(+) high-grade cervical lesions. Therefore, the CervicGen HPV RT-qDX assay showed a significantly higher sensitivity (91.1%) compared to the real-time NASBA assay (41.1%). In normal cytohistology cases, the specificity was 98.6% and 53.7% for HPV mRNA RT-qPCR and HPV DNA testing, respectively. These results demonstrate that HPV mRNA RT-qPCR better reflects clinical diagnosis. In conclusion, it is suggested that HPV mRNA RT-qPCR overcomes the shortcomings of lower specificity seen in the DNA assay and the lower sensitivity of the commercialized HPV mRNA real-time NASBA assay when testing from ThinPrep Pap samples.
authors.affiliation: Department of Pathology, Yonsei University Wonju College of Medicine, Wonju, Gangwon 220-701, Republic of Korea.
authors.forename: Jijgee
authors.initials: J
authors.lastname: Munkhdelger
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Gangwon 220-710, Republic of Korea.
authors.forename: Geehyuk
authors.initials: G
authors.lastname: Kim
authors.affiliation: M&D, Inc., Wonju Eco Environmental Technology Center, Wonju, Gangwon 200-722, Republic of Korea.
authors.forename: Hye-young
authors.initials: HY
authors.lastname: Wang
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Gangwon 220-710, Republic of Korea; Department of Clinical Laboratory Science, Hyejeon College, Hongseong, Chungnam 350-702, Republic of Korea.
authors.forename: Dongsup
authors.initials: D
authors.lastname: Lee
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Gangwon 220-710, Republic of Korea; Institute for Life Science and Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Sunghyun
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Biomedical Laboratory Science, Songho College, Hoengseong, Gangwon 225-704, Republic of Korea.
authors.forename: Yeonim
authors.initials: Y
authors.lastname: Choi
authors.affiliation: Institute of Lifestyle Medicine, Wonju College of Medicine, Yonsei University, Wonju, Gangwon 220-701, Republic of Korea.
authors.forename: Eunhee
authors.initials: E
authors.lastname: Choi
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Gangwon 220-710, Republic of Korea.
authors.forename: Sunyoung
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Pusan.
authors.forename: Hyunwoo
authors.initials: H
authors.lastname: Jin
authors.affiliation: Department of Pathology, Yonsei University Wonju College of Medicine, Wonju, Gangwon 220-701, Republic of Korea. Electronic address: abba@yonsei.ac.kr.
authors.forename: Kwang Hwa
authors.initials: KH
authors.lastname: Park
authors.affiliation: M&D, Inc., Wonju Eco Environmental Technology Center, Wonju, Gangwon 200-722, Republic of Korea. Electronic address: hyelee@yonsei.ac.kr.
authors.forename: Hyeyoung
authors.initials: H
authors.lastname: Lee
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D005260:Female; D061809:Human Papillomavirus DNA Tests / Q000379:methods; D006801:Humans; D008875:Middle Aged; D009856:Oncogene Proteins, Viral / Q000235:genetics; D065006:Papanicolaou Test / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis*
chemical_list: D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger
keywords: HPV E6/E7 mRNA RT-qPCR; Human papillomavirus (HPV); Molecular diagnosis; Real-time NASBA