medigraphic.com
ISSN 1561-3054 (Electronic)

2014, Number 3

<< Back Next >>

Rev Cubana Med Trop 2014; 66 (3)

Standardization of immunochemical method for detection of type 16 human papilomavirus in cervix uterine lesions

Torres LG, Soto BY, Capó PV, López FLX, de la Torre JAI, Goicolea MA, Limonta VD, Pérez RA, Govín CA, Morier DL, Caballero LY, Rodríguez SH, Álvarez GRI, Calvo PA, Escobar PX, Limia LCM, Kourí CV

Language: Spanish
References: 27
Page: 433-446
PDF size: 271.49 Kb.


ABSTRACT

Introduction: Human Papillomavirus (HPV) infection is the necessary condition for the occurernce and development of cervical cancer. The high oncogenic risk genotypes are the responsible for this type of neoplasia and the most frequent is HPV 16 that affects roughly 60 % of cases. Commercial kits for HPV detection are expensive for resource-poor countries, which suggests the search for alternative throguh non-expensive simple protocoles.
Objectives: to standardize an immunochemical method for the detection of HPV 16 L1 antigen in cervical samples of patients with squamous intraepithelial lesions and to determine the diagnostic coincidence between the immunochemical method and the real-time polymerase chain reaction to estimate the usefulness of this method for the detection of cervical infection with this viral genotype.
Methods: three immunostaining methods (Two-Step Indirect Immunoperoxidase, Labelled Streptavidin-Biotin and Enhanced Polymer) were compared in terms of analytical sensitivity, nonspecific background staining and time of completion, for the detection of protein L1 of HPV-16 in a cell line derived from human cervical carcinoma and clinical samples from uterine cervix. The optimized protocol was applied to 82 cervical samples from women aged 30-59 years with squamous intraepithelial lesions and to 10 samples of sexually active women without previous signals of positive cytology. The presence of type 16 HPV was also detected with the aid of RT-PCR.
Results: the Streptavidin-Biotin system was the most sensitive and specific. The diagnostic agreement between the immunochemical method and the real-time polymerase chain reaction reached 98.6 %, sensitivity was 98.57 % and specificity was 91.67 %, with positive and negative predictive values above 90 %.
Conclusions: the validity of the immunochemical method as a confirmatory test for infection by HPV-16 has been demonstrated. The normalized immunochemical method proved to be a sensitive, simple, relatively fast method to detect HPV from clinical samples of cervical cells. Furthermore, this method provides information quickly, avoiding the use of invasive methods in patients.


REFERENCES

  1. zur Hausen H. Papillomaviruses in the causation of human cancers - a brief historical account. Virology. 2009;384(2):260-5. Epub 2009/01/13.

  2. De Vuyst H, Clifford GM, Nascimento MC, Madeleine MM, Franceschi S. Prevalence and type distribution of human papillomavirus in carcinoma and intraepithelial neoplasia of the vulva, vagina and anus: a meta-analysis. Int J Cancer. 2009;124(7):1626-36. Epub 2008/12/31.

  3. Coste J, Cochand-Priollet B, de Cremoux P, Le Gales C, Cartier I, Molinie V, et al. Cross sectional study of conventional cervical smear, monolayer cytology, and human papillomavirus DNA testing for cervical cancer screening. BMJ. 2003;326(7392):733. Epub 2003/04/05.

  4. Murillo R, Almonte M, Pereira A, Ferrer E, Gamboa OA, Jeronimo J, et al. Cervical cancer screening programs in Latin America and the Caribbean. Vaccine. 2008;26 Suppl 11:L37-48. Epub 2008/11/26.

  5. Bess S, Gran MA, Torres RM, Alonso I, Lopez LM, Velazquez JA, et al. Anuario Estadístico de Salud 2012. Especial ed. La Habana: Oficina Nacional de Estadísticas e Información de la Dirección Nacional de Registros Médicos del Ministerio de Salud Pública de Cuba; 2013. p. 1-205.

  6. Boenisch T. Staining Methods. In: Boenisch T, editor. Immunochemical Staining Methods H A N D B O O K 3rd ed. California, USA: DakoCytomation - Carpinteria, CA, USA; 2001. p. 26-31.

  7. Soto Y, Kourí V, Martínez PA, Correa C, Torres G, Goicolea A, et al. Standardization of a real-time based polimerase chain reaction system for the quantification of human papillomavirus of high oncogenic risk. Vaccimonitor. 2012; 21(1):30-7.

  8. Soto Y, Mune M, Morales E, Goicolea A, Mora J, Sanchez L, et al. Human Papillomavirus infections in Cuban women with cervical intraepithelial neoplasia. Sex Transm Dis. 2007;34(12):974-6.

  9. Solomon D, Davey D, Kurman R, Moriarty A, O’Connor D, Prey M, et al. The 2001 Bethesda System. Terminology for Reporting Results of Cervical Cytology. JAMA. 2002;287(16):2114-9. Epub 2001. Forum Group Members and the Bethesda 2001 Workshop.

  10. Schmitz M, Scheungraber C, Herrmann J, Teller K, Gajda M, Runnebaum IB, et al. Quantitative multiplex PCR assay for the detection of the seven clinically most relevant high-risk HPV types. J Clin Virol. 2009;44(4):302-7. Epub 2009/02/19.

  11. Kiernan JA. Immunohistochemistry. In: Kiernan JA, editor. Histological and histochemical methods Theory and practice. Oxford, UK Scion Publishing Limited; 2010. p. 483-519.

  12. Hernandez J, Elahi A, Siegel E, Coppola D, Riggs B, Shibata D. HPV L1 Capsid Protein Detection and Progression of Anal Squamous Neoplasia. Am J Clin Pathol. 2011;135:436-41.

  13. Torres G IF, Capó V, López LX, López MC, Limonta D, Fenoy S, et al. Genital microsporidiosis in women with AIDS: A post-mortem study. Rev Iberoam Micol. 2013;30(1):47-50.

  14. Jara D, Rengifo E, Seminario V, Malaspina MF, Yamada A, Vitorio J. Immunohistochemical detection of Human Papillomavirus in cervical cancer. Anales de la Facultad de Medicina. 2000;61(1):31-6.

  15. Kosel S, Burggraf S, Engelhardt W, Olgemoller B. Correlation of HPV16 L1 capsid protein expression in cervical dysplasia with HPV16 DNA concentration, HPV16 E6*I mRNA and histologic outcome. Acta Cytol. 2009;53(4):396-401. Epub 2009/08/25.

  16. McLean CS, Churcher MJ, Meinke J, Smith GL, Higgins G, Stanley M, et al. Production and characterisation of a monoclonal antibody to human papillomavirus type 16 using recombinant vaccinia virus. J Clin Pathol. 1990;43(6):488-92. Epub 1990/06/01.

  17. Banks L, Spence P, Androphy E, Hubbert N, Matlashewski G, Murray A, et al. Identification of human papillomavirus type 18 E6 polypeptide in cells derived from human cervical carcinomas. J Gen Virol. 1987;68(5):1351-9. Epub 1987/05/01.

  18. Holmes BJ, Westra WH. The expanding role of cytopathology in the diagnosis of HPV-related squamous cell carcinoma of the head and neck. Diagn Cytopathol. 2014;42(1):85-93. Epub 2013/06/12.

  19. Lawson JS, Glenn WK, Heng B, Ye Y, Tran B, Lutze-Mann L, et al. Koilocytes indicate a role for human papilloma virus in breast cancer. Br J Cancer. 2009;101(8):1351-6. Epub 2009/09/24.

  20. Griesser H, Sander H, Hilfrich R, Moser B, Schenck U. Correlation of immunochemical detection of HPV L1 capsid protein in pap smears with regression of high-risk HPV positive mild/moderate dysplasia. Anal Quant Cytol Histol. 2004;26(5):241-5. Epub 2004/11/25.

  21. Hoshikawa S, Sano T, Yoshida T, Ito H, Oyama T, Fukuda T. Immunohistological analysis of HPV L1 capsid protein and p16 protein in low-grade dysplastic lesions of the uterine cervix. Pathol Res Pract. 2010;206(12):816-20. Epub 2010/10/23.

  22. van Bogaert LJ. Cervical preneoplasia biomarkers: a conundrum for the community based gynecologic surgical pathologist. J Gynecol Oncol. 2014;25(1):3- 5. Epub 2014/01/25.

  23. Norman I, Hjerpe A, Andersson S. High-risk HPV L1 capsid protein as a marker of cervical intraepithelial neoplasia in high-risk HPV-positive women with minor cytological abnormalities. Oncol Rep. 2013;30(2):695-700. Epub 2013/06/13.

  24. Rashed MM, Bekele A. The prevalence and pattern of HPV-16 immunostaining in uterine cervical carcinomas in Ethiopian women: a pilot study. Pan Afr Med J. 2011;8:21. Epub 2011/11/29.

  25. Xiao W, Bian ML, Ma L, Liu J, Chen Y. [Detection of human papillomavirus L1 capsid protein expression in liquid-based cytology samples with abnormal cytology.]. Zhonghua Fu Chan Ke Za Zhi. 2009;44(12):887-91. Epub 2010/03/03.

  26. Sarmadi S, Izadi-mood N, Pourlashkari M, Yarandi F, Sanii S. HPV L1 capsid protein expression in squamous intraepithelial lesions of cervix uteri and its relevance to disease outcome. Arch Gynecol Obstet. 2011;285(3):779-84. Epub 2011/07/27.

  27. Choi YS, Kang WD, Kim SM, Choi YD, Nam JH, Park CS, et al. Human papillomavirus L1 capsid protein and human papillomavirus type 16 as prognostic markers in cervical intraepithelial neoplasia 1. Int J Gynecol Cancer. 2010;20(2):288-93. Epub 2010/02/06.




2020     |     www.medigraphic.com