, Volume 30, Issue 3, pp 387–393 | Cite as

Association of HPV16 and 18 genomic copies with histological grades of cervical lesions

  • Negar Joharinia
  • Ali Farhadi
  • Seyed Younes Hosseini
  • Akbar Safaei
  • Jamal SarvariEmail author
Original Article


The possible correlation between HPV16 and HPV18 genomic copies with the grade of cervical lesions needs more investigations. The aim of this study was to quantify genomic copies of HPV16 and 18 simultaneously and to find out the correlation between genomic copies numbers and different grades of lesions. Therefore, a total of 102 formalin-fixed and paraffin-embedded tissue specimens, 33 LSILI, 43 HSIL, and 26 squamous/adenocarcinoma were subjected to DNA extraction. The β-globin gene was selected to qualify the extracted DNA as well as normalization of viral titers using Taq-Man real-time PCR. The presence HPV16 and/or 18 were screened in tissue samples by nested PCR method, then an in- house Taq-Man Duplex real-time PCR assay was employed to quantify their genomic copies. The mean age of participants was 43 ± 13. Out of 102, 80 samples were positive for HPV16 and/or 18 DNA. There was a statistically significant association between HPV16 genomic copies and progression of cervical lesions (P < 0.001). In contrast, no such an association was found in the case of HPV18 (P = 0.51). Moreover, with 95% confidence intervals, 2.3–4 genomic copies of HPV16 genome/cell could be applicable to distinguish LSIL from HSIL and SCC. In conclusion, quantification of HPV16 genomic copy number showed a close association with progression of cervical lesion. Furthermore, HPV16 genomic copies of 4 copies/cell could be a set point to differentiate LSIL from HSIL.


HPV16 and HPV18 LSIL HSIL SCC Taq-Man real-time PCR Genomic copies 



The present study was extracted from the thesis written by Negar Joharinia, and was financially supported by Shiraz University of Medical Sciences (Grant No: 94-10934). Special thanks to Prof. Abbas Behzad-Behbahani and the staff of Diagnostic Laboratory Sciences and Technology Research Center for their assistance. The authors wish to thank Mr. H. Argasi at the Research Consultation Center (RCC) of Shiraz University of Medical Sciences for his invaluable assistance in editing this manuscript.

Compliance with ethical standards

Conflict of interest

All the authors declare that they have no conflict of interest.


  1. 1.
    Bakhtiarizadeh S, et al. Almost complete lack of human cytomegalovirus and human papillomaviruses genome in benign and malignant breast lesions in Shiraz, Southwest of Iran. Asian Pac J Cancer Prev. 2017;9(12):3319–24.Google Scholar
  2. 2.
    Barbosa M, Schlegel RJO. The E6 and E7 genes of HPV-18 are sufficient for inducing two-stage in vitro transformation of human keratinocytes. Oncogene. 1989;4(12):1529–32.PubMedGoogle Scholar
  3. 3.
    Briolat J, et al. HPV prevalence, viral load and physical state of HPV-16 in cervical smears of patients with different grades of CIN. Int J Cancer. 2007;121(10):2198–204.CrossRefGoogle Scholar
  4. 4.
    Broccolo F, et al. Prevalence and viral load of oncogenic human papillomavirus types associated with cervical carcinoma in a population of North Italy. J Med Virol. 2009;81(2):278–87.CrossRefGoogle Scholar
  5. 5.
    Burd EM. Human papillomavirus and cervical cancer. Clin Microbiol Rev. 2003;16(1):1–17.CrossRefGoogle Scholar
  6. 6.
    Cancer, I.I.C.o.H.a. Human papillomavirus and related diseases report. ICO Information Centre on HPV and Cancer, 2015.Google Scholar
  7. 7.
    Carcopino X, et al. Significance of HPV 16 and 18 viral load quantitation in women referred for colposcopy. J Med Virol. 2012;84(2):306–13.CrossRefGoogle Scholar
  8. 8.
    Carrillo-García A, et al. Impact of human papillomavirus coinfections on the risk of high-grade squamous intraepithelial lesion and cervical cancer. Gynecol Oncol. 2014;134(3):534–9.CrossRefGoogle Scholar
  9. 9.
    de Jesus SP, et al. A high prevalence of human papillomavirus 16 and 18 co-infections in cervical biopsies from southern Brazil. Braz J Microbiol. 2018;49:220–3.CrossRefGoogle Scholar
  10. 10.
    Del Río-Ospina L, et al. The DNA load of six high-risk human papillomavirus types and its association with cervical lesions. BMC Cancer. 2015;15(1):100.CrossRefGoogle Scholar
  11. 11.
    Dickson EL, et al. Cervical cytology and multiple type HPV infection: a study of 8182 women ages 31–65. Gynecol Oncol. 2014;133(3):405–8.CrossRefGoogle Scholar
  12. 12.
    Dong L, et al. Human papillomavirus viral load as a useful triage tool for non-16/18 high-risk human papillomavirus positive women: a prospective screening cohort study. Gynecol Oncol. 2018;148(1):103–10.CrossRefGoogle Scholar
  13. 13.
    Dong B, et al. Type-specific high-risk human papillomavirus viral load as a viable triage indicator for high-grade squamous intraepithelial lesion: a nested case- control study. Cancer Manag Res. 2018;10:4839–51.CrossRefGoogle Scholar
  14. 14.
    Eghbali SS, et al. Oncogenic human papillomavirus genital infection in southern Iranian women: population-based study versus clinic-based data. Virol J. 2012;9(1):194.CrossRefGoogle Scholar
  15. 15.
    Farhadi A, et al. High-risk human papillomavirus infection in different histological subtypes of renal cell carcinoma. J Med Virol. 2014;86(7):1134–44.CrossRefGoogle Scholar
  16. 16.
    Fitzmaurice C, et al. Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-adjusted life-years for 32 cancer groups, 1990 to 2015: a systematic analysis for the global burden of disease study. JAMA Oncol. 2017;3(4):524–48.CrossRefGoogle Scholar
  17. 17.
    Flores R, et al. Cross-sectional analysis of oncogenic HPV viral load and cervical intraepithelial neoplasia. Int J Cancer. 2006;118(5):1187–93.CrossRefGoogle Scholar
  18. 18.
    Gao G, Smith DI. Human papillomavirus and the development of different cancers. Cytogenet Genome Res. 2017;150:185–93.CrossRefGoogle Scholar
  19. 19.
    Guan P, et al. Human papillomavirus types in 115,789 HPV-positive women: a meta-analysis from cervical infection to cancer. Int J Cancer. 2012;131(10):2349–59.CrossRefGoogle Scholar
  20. 20.
    Hesselink AT, et al. High-risk human papillomavirus DNA load in a population-based cervical screening cohort in relation to the detection of high-grade cervical intraepithelial neoplasia and cervical cancer. Int J Cancer. 2009;124(2):381–6.CrossRefGoogle Scholar
  21. 21.
    Ho CM, et al. Type-specific human papillomavirus oncogene messenger RNA levels correlate with the severity of cervical neoplasia. Int J Cancer. 2010;127(3):622–32.CrossRefGoogle Scholar
  22. 22.
    Khorasanizadeh F, et al. Epidemiology of cervical cancer and human papilloma virus infection among Iranian women—analyses of national data and systematic review of the literature. Gynecol Oncol. 2013;128(2):277–81.CrossRefGoogle Scholar
  23. 23.
    Kiani SJ, et al. Detection and typing of human papilloma viruses by nested multiplex polymerase chain reaction assay in cervical cancer. Jundishapur J Microbiol. 2015;8(12):e26441.Google Scholar
  24. 24.
    Mahmoodvan S, et al. No detection of Streptococcus g allolyticus and Helicobacter pylori in colorectal cancer tissue samples in Shiraz. Iran. Iran J Cancer Prev. 2017;10(1):e6337.Google Scholar
  25. 25.
    Mahmoudvand S, et al. Presence of human papillomavirus DNA in colorectal cancer tissues in Shiraz, Southwest Iran. Asian Pac J Cancer Prev APJCP. 2014;16(17):7883–7.CrossRefGoogle Scholar
  26. 26.
    Mayrand M-H, et al. Human papillomavirus DNA versus Papanicolaou screening tests for cervical cancer. N Engl J Med. 2007;357(16):1579–88.CrossRefGoogle Scholar
  27. 27.
    Mittal S, et al. Risk of high-grade precancerous lesions and invasive cancers in high-risk HPV-positive women with normal cervix or CIN 1 at baseline—a population-based cohort study. Int J Cancer. 2017;140(8):1850–9.CrossRefGoogle Scholar
  28. 28.
    Ramírez-Flores M, et al. HPV 16 and 18 viral loads are greater in patients with high-grade cervical epithelial lesions. Eur J Gynaecol Oncol. 2016;37(5):644–8.PubMedGoogle Scholar
  29. 29.
    Serrano B, et al. Epidemiology and burden of HPV-related disease. Best Pract Res Clin Obstet Gynaecol. 2017;47:14–26.CrossRefGoogle Scholar
  30. 30.
    Villa LL, Schlegel RJV. Differences in transformation activity between HPV-18 and HPV-16 map to the viral LCR-E6-E7 region. Virology. 1991;181(1):374–7.CrossRefGoogle Scholar
  31. 31.
    Wu Z, et al. Association between human papillomavirus (HPV) 16, HPV18, and other HR-HPV viral load and the histological classification of cervical lesions: results from a large-scale cross-sectional study. J Med Virol. 2017;89(3):535–41.CrossRefGoogle Scholar

Copyright information

© Indian Virological Society 2019

Authors and Affiliations

  • Negar Joharinia
    • 1
  • Ali Farhadi
    • 2
  • Seyed Younes Hosseini
    • 1
  • Akbar Safaei
    • 3
  • Jamal Sarvari
    • 1
    • 4
    Email author
  1. 1.Department of Bacteriology and Virology, School of MedicineShiraz University of Medical SciencesShirazIran
  2. 2.Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical SciencesShiraz University of Medical SciencesShirazIran
  3. 3.Department of Pathology, School of MedicineShiraz University of Medical SciencesShirazIran
  4. 4.Gastroenterohepatology Research CenterShiraz University of Medical SciencesShirazIran

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