Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
권호 표지
DOI QR코드

DOI QR Code

Analytical Performance of Sensitivity and Specificity for Rapid Multiplex High Risk Human Papillomavirus Detection Kit: HPV ViroCheck

고위험군 HPV 검출을 위한 분석적 민감도와 특이도 성능평가

  • Park, Sunyoung (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Yoon, Hyeonseok (Optipharm M&D, Inc., Wonju Eco Environmental Technology Center) ;
  • Bang, Hyeeun (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Kim, Yeun (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Choi, Seongkyung (Optipharm M&D, Inc., Wonju Eco Environmental Technology Center) ;
  • Ahn, Sungwoo (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Kim, Jungho (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Lee, Suji (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Yang, Ji Yeong (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Lee, Dongsup (Department of Clinical Laboratory Science, Hyejeon College)
  • 박선영 (연세대학교 임상병리학과) ;
  • 윤현석 (옵티팜 엠엔디) ;
  • 방혜은 (연세대학교 임상병리학과) ;
  • 김연 (연세대학교 임상병리학과) ;
  • 최성경 (옵티팜 엠엔디) ;
  • 안성우 (연세대학교 임상병리학과) ;
  • 김정호 (연세대학교 임상병리학과) ;
  • 이수지 (연세대학교 임상병리학과) ;
  • 양지영 (연세대학교 임상병리학과) ;
  • 이동섭 (혜전대학교 임상병리과)
  • Received : 2017.10.12
  • Accepted : 2017.11.01
  • Published : 2017.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Human papillomaviruses (HPVs) are major causes of cervical cancer. Sixteen high risk HPVs, including HPV 16, HPV 18, HPV31, HPV 33, HPV 35, HPV 39, HPV 45, HPV 51, HPV 52, HPV 53, HPV 56, HPV 58, HPV 59, HPV 66, HPV 68, and HPV 69 are found in cervical cancer. HPVs 16 and 18 are mainly presented in 70% of cervical cancer. Therefore, identifying the presence of these high-risk HPVs is crucial. The objective of this study is to establish the HPV ViroCheck for detecting 16 HR-HPVs and genotypes of HPVs 16 and 18, as well as to analyze the analytical performance of HPV ViroCheck. We performed the analytical sensitivity of HPV E6 / E7 genes of 16 high risk HPVs to confirm the limit of detection. Then, a cross reactivity of HPV ViroCheck with microorganisms and viruses related to the cervix were analyzed for analytical specificity. Analytical sensitivity of high risk HPV genotypes ranged from 1 to 100 copies when using cloned DNAs. The limit of detection was 10 cells for both SiHa and HeLa cells. Cervical-related microorganisms and viruses did not show cross-reactivity to HPV DNA. Moreover, the intra- and inter-assay coefficient variations (CVs) were below 5%. In conclusion, HPV Virocheck will be useful for the detection of 16 HR HPVs, as well as HPV 16 and HPV 18 genotypes rapidly.

인간유두종 바이러스 (HPV)는 자궁경부암의 주요 원인이며, 자궁경부암의 주요 원인 바이러스는 16종의 고위험군 유전형 HPV 16, HPV 18, HPV31, HPV 33, HPV 35, HPV 39, HPV 45, HPV 51, HPV 52, HPV 53, HPV 56, HPV 58, HPV 59, HPV 66, HPV 68, HPV 69 이다. 특히, HPV 16형과 HPV 18형이 HPV 양성 암환자의 70%에서 발견된다. 따라서, 바이러스의 존재 유무를 확인하는 것은 환자의 스크리닝에 도움을 주며, 최근에 세포학적 검사와 함께 보조적인 검사법으로 사용되고 있다. 본 연구는 16종의 고위험군 바이러스와 HPV 16, HPV 18 유전형을 검출 할 수 있는 HPV ViroCheck의 발암 유전자의 분석 성능을 확립하기 위한 목적으로 수행되었다. 먼저, 16종의 고위험군 HPV의 발암유전자 E6/E7 유전형의 검출한계를 확인하기 위하여, 분석적 민감도를 수행하였다. 그리고, 관련된 미생물 및 바이러스에서의 교차반응 및 정확도를 비교하여 평가하였다. 고위험군 HPV 유전자형의 민감도는 Clone DNA를 이용 하였을 때, 최대 1카피에서 100 카피까지 검출이 가능하였고, SiHa 세포와 Hela 세포의 경우 최소 10 세포까지 검출이 가능하였다. 자궁경부 관련 미생물 및 바이러스에서 HPV 유전형에 대한 교차 반응은 나타내지 않았다. 또한, 측정법 내 변동계수 및 측정법 간 변동계수 실험 결과 변동계수가 5% 이하로 정확도가 높았다. 위의 분석 성능자료는 HPV ViroCheck의 유전자형 검사의 식품의약품 안전처의 체외진단용 의료기기 허가를 위한 자료로 사용 될 것이며, 향후, HPV 16종의 발암유전자 검출과 HPV 16, HPV 18 유전형 검출 연구에 도움이 될 것이다.

Keywords

References

  1. Forman D, de Martel C, Lacey CJ, Soerjomataram I, Lortet-Tieulent J, Bruni L, et al. Global burden of human papillomavirus and related diseases. Vaccine. 2012;30(Suppl 5):F12-23. https://doi.org/10.1016/j.vaccine.2012.07.055
  2. Munoz N, Bosch FX, de Sanjose S, Herrero R, Castellsague X, Shah KV, et al. Epidemiologic classification of human papillomavirus types associated with cervical cancer. N Engl J Med. 2003;348(6):518-527. https://doi.org/10.1056/NEJMoa021641
  3. Kjaer SK, Munk C, Junge J, Iftner T. Carcinogenic HPV prevalence and age-specific type distribution in 40,382 women with normal cervical cytology, ASCUS/LSIL, HSIL, or cervical cancer: what is the potential for prevention? Cancer Causes Control. 2014;25(2):179-189. https://doi.org/10.1007/s10552-013-0320-z
  4. Castle PE, Jeronimo J, Temin S, Shastri SS, et al. Screening to prevent invasive cervical cancer: ASCO resource-stratified clinical practice guideline. J Clin Oncol. 2017;35(11):1250-1252. https://doi.org/10.1200/JCO.2016.71.6563
  5. Castle PE, Katki HA. Benefits and risks of HPV testing in cervical cancer screening. Lancet Oncol. 2010;11(3):214-215. https://doi.org/10.1016/S1470-2045(09)70385-7
  6. Herbert A. Primary HPV testing: a proposal for co-testing in initial rounds of screening to optimise sensitivity of cervical cancer screening. Cytopathology. 2017;28(1):9-15. https://doi.org/10.1111/cyt.12334
  7. Min KJ, Lee YJ, Suh M, Yoo CW, Lim MC, Choi J, et al. The Korean guideline for cervical cancer screening. J Gynecol Oncol. 2015;26(3):232-239. https://doi.org/10.3802/jgo.2015.26.3.232
  8. Lee JK, Hong JH, Kang S, Kim DY, Kim BG, Kim SH, et al. Practice guidelines for the early detection of cervical cancer in Korea: Korean Society of Gynecologic Oncology and the Korean Society for Cytopathology 2012 edition. J Gynecol Oncol. 2013;24(2):186-203. https://doi.org/10.3802/jgo.2013.24.2.186
  9. Ratnam S, Coutlee F, Fontaine D, Bentley J, Escott N, Ghatage P, et al. Clinical performance of the PreTect HPV-Proofer E6/E7 mRNA assay in comparison with that of the Hybrid Capture 2 test for identification of women at risk of cervical cancer. J Clin Microbiol. 2010;48(8):2779-2785. https://doi.org/10.1128/JCM.00382-10
  10. Park SM, Lee SK, Kim YS. Inhibition of cervical cancer cell growth by gene silencing of HPV16 E6 Induced by short-interfering RNA. Korean J Clin Lab Sci. 2011;43(3):89-97.
  11. Thomas M, Pim D, Banks L. The role of the E6-p53 interaction in the molecular pathogenesis of HPV. Oncogene. 1999; 18(53):7690-7700. https://doi.org/10.1038/sj.onc.1202953
  12. Storey A, Thomas M, Kalita A, Harwood C, Gardiol D, Mantovani F, et al. Role of a p53 polymorphism in the development of human papillomavirus-associated cancer. Nature. 1998;393(6682):229-234. https://doi.org/10.1038/30400
  13. Caldeira S, Dong W, Tommasino M. Analysis of E7/Rb associations. Methods Mol Med. 2005;119:363-379.
  14. Lee JB, Park CE. Investigation of detected by recent various human papillomavirus from general hospital in Seoul area. Korean J Clin Lab Sci. 2016;48(3):247-254. https://doi.org/10.15324/kjcls.2016.48.3.247
  15. Munkhdelger J, Kim GH, Wang HY, Lee DS, Kim SH, Choi YI, et al. Performance of HPV E6/E7 mRNA RT-qPCR for screening and diagnosis of cervical cancer with Thinprep pap test samples. Exp Mol Pathol. 2014;97(2):279-284. https://doi.org/10.1016/j.yexmp.2014.08.004
  16. Kim JH, Wang HY, Kim SY, Park SD, Yu KM, Kim HY, Uh Y, Lee HY. Evaluation of the Punch-$it^{TM}$ NA-Sample kit for detecting microbial DNA in blood culture bottles using PCR-reverse blot hybridization assay. Journal of Microbiological Methods. 2016; 128:24-30. https://doi.org/10.1016/j.mimet.2016.06.001
  17. Walsh PS, Metzger DA, Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques. 1991;10(4):506-513.
  18. Mincheva A, Gissmann L, zur Hausen H. Chromosomal integration sites of human papillomavirus DNA in three cervical cancer cell lines mapped by in situ hybridization. Med Microbiol Immunol. 1987;176(5): 245-256. https://doi.org/10.1007/BF00190531
  19. Terry G, Ho L, Londesborough P, Cuzick J, Mielzynska-Lohnas I, Lorincz A. Detection of high-risk HPV types by the hybrid capture 2 test. J Med Virol. 2001;65(1):155-162. https://doi.org/10.1002/jmv.2015
  20. Du Chateau BK, Schroeder ER, Munson E. Clinical laboratory experience with cervista HPV HR as a function of cytological classification: comparison with retrospective digene HC2 high-risk HPV DNA test data. J Clin Microbiol. 2013;51(3): 1057-1058. https://doi.org/10.1128/JCM.02836-12
  21. Zhou F, Pulinthanathu R, Elgert P, Cangiarella J, Simsir A. Sensitivity of high-risk HPV Hybrid Capture II (hrHPV HC2) test using SurePath(TM) specimens in the prediction of cervical high-grade squamous lesions. Diagn Cytopathol. 2015;43(5): 381-387. https://doi.org/10.1002/dc.23243
  22. Huijsmans CJ, Geurts-Giele WR, Leeijen C, Hazenberg HL, van Beek J, de Wild C, et al. HPV prevalence in the Dutch cervical cancer screening population (DuSC study): HPV testing using automated HC2, cobas and aptima workflows. BMC Cancer. 2016;16:922. https://doi.org/10.1186/s12885-016-2961-2
  23. Kash N, Lee MA, Kollipara R, Downing C, Guidry J, Tyring SK. Safety and efficacy data on vaccines and immunization to human papillomavirus. J Clin Med. 2015;4(4):614-633. https://doi.org/10.3390/jcm4040614
  24. Yang A, Farmer E, Wu TC, Hung CF. Perspectives for therapeutic HPV vaccine development. J Biomed Sci. 2016;23:75. https://doi.org/10.1186/s12929-016-0293-9
  25. Kim TJ, Jin HT, Hur SY, Yang HG, Seo YB, Hong SR, et al. Clearance of persistent HPV infection and cervical lesion by therapeutic DNA vaccine in CIN3 Patients. Nat Commun. 2014;30:5317.
  26. Trimble CL, Morrow MP, Kraynyak KA, Shen X, Dallas M, Yan J, et al. Safety, efficacy, and immunogenicity of VGX-3100, a therapeutic synthetic DNA vaccine targeting human papillomavirus 16 and 18 E6 and E7 proteins for cervical intraepithelial neoplasia 2/3: a randomised, double-blind, placebocontrolled phase 2b trial. Lancet. 2015;386(10008):2078-2088. https://doi.org/10.1016/S0140-6736(15)00239-1
  27. Morrow MP, Kraynyak KA, Sylvester AJ, Shen X, Amante D, Sakata L, et al. Augmentation of cellular and humoral immune responses to HPV16 and HPV18 E6 and E7 antigens by VGX-3100. Mol Ther Oncolytics. 2016;3:16025. https://doi.org/10.1038/mto.2016.25

Cited by

  1. Diagnostic Availability of Estrogen Receptor Alpha mRNA on Cervical Cancer Tissue vol.50, pp.4, 2017, https://doi.org/10.15324/kjcls.2018.50.4.449
  2. Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus vol.51, pp.3, 2019, https://doi.org/10.15324/kjcls.2019.51.3.309