Human papillomavirus detection in cervical dysplasias or neoplasias and in condylomata acuminaata by in situ hybridization with biotinylated DNA probes

Guimarães, Eliane Machado; Brasileiro Filho, Geraldo; Pena, Sérgio Danilo Junho

doi:10.1590/S0036-46651992000400007

Abstracts

Specimens from cervical dysplasias or carcinomas and genital condylomata acuminata were retrospectively analysed by in situ hybridization (ISH) with bioti-nylated DNA probes for human papillomavirus (HPV) types 6, 11, 16 and 18. In the control group no case was positive for HPV DNA. In mild/moderate dysplasias, 4 cases (14%) were positive for HPV 6 or 11 and 2 cases (7%), for HPV 16. In the severe dysplasia/in situ carcinoma group, 9 cases (31%) showed presence of DNA of HPV types 16 or 18. Six invasive carcinomas (20%) were positive for HPV type 16 or 18. Among condylomata acuminata, 22 cases (73%) were positive for HPV types 6 or 11. In all ISH-positive cases only one viral type was detected. No correlation between HPV DNA positivity and histological findings of HPV infection was observed. Although less sensitive than some other molecular biology techniques, in situ hybridization with biotinylated DNA probes proved to be simple and useful for detecting and typing HPV in samples routinely received for histopathological analysis.

Human papillomavirus; In situ hybridization; Cervical dysplasia; Cervical cancer; Condylomata acuminata

Amostras de displasias ou carcinomas do colo uterino e de condilomas acuminados da região genital foram analisados retrospectivamente por hibridização in situ (HIS) com sondas de DNA biotiniladas de papilomavirus humano (HPV) tipos 6, 11, 16 e 18. Nenhum caso do grupo controle foi positivo para DNA de HPV. Em displasias leve/moderada, 4 casos (14%) foram positivos para HPV 6 ou 11 e 2 casos (7%), para HPV 16. No grupo de displasia acentuada/carcinoma in situ, 9 casos (31%) tinham DNA de HPV tipos 16 ou 18. Seis carcinomas invasores (20%) foram positivos para HPV tipos 16 ou 18. Entre os condilomas acuminados, 22 casos (73%) foram positivos para HPV tipos 6 ou 11. Em todos os casos positivos pela HIS somente um tipo viral foi encontrado. Não foi observada correlação entre a positividade para DNA de HPV e achados histológicos de infecção por HPV. Apesar de menos sensível que algumas outras técnicas de biologia molecular, a hibridização in situ com sondas de DNA biotiniladas mostrou-se simples e adequada para detecção e tipagem de HPV em amostras enviadas rotineiramente para estudo histopatológico.

ORIGINAL ARTICLES

Human papillomavirus detection in cervical dysplasias or neoplasias and in condylomata acuminaata by in situ hybridization with biotinylated DNA probes

Detecção de papilomavirus humano em displasias ou neoplasias cervicais e em condilomas acuminados por hibridização in situ com sondas de DNA biotiniladas

Eliane Machado Guimarães^I; Geraldo Brasileiro Filho^I; Sérgio Danilo Junho Pena^II

^IDepartamento de Anatomia Patológica e Medicina Legal, Faculdade de Medicina, UFMG. Belo Horizonte, MG, Brasil

^IIDepartamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, UFMG. Belo Horizonte, MG, Brasil

^{Address for correspondence} Address for correspondence: Prof. Geraldo Brasileiro Filho Departamento de Anatomia Patológica Faculdade de Medicina UFMG Caixa Postal, 340 30.130, Belo Horizonte-MG, Brasil

SUMMARY

Specimens from cervical dysplasias or carcinomas and genital condylomata acuminata were retrospectively analysed by in situ hybridization (ISH) with bioti-nylated DNA probes for human papillomavirus (HPV) types 6, 11, 16 and 18. In the control group no case was positive for HPV DNA. In mild/moderate dysplasias, 4 cases (14%) were positive for HPV 6 or 11 and 2 cases (7%), for HPV 16. In the severe dysplasia/in situ carcinoma group, 9 cases (31%) showed presence of DNA of HPV types 16 or 18. Six invasive carcinomas (20%) were positive for HPV type 16 or 18. Among condylomata acuminata, 22 cases (73%) were positive for HPV types 6 or 11.

In all ISH-positive cases only one viral type was detected. No correlation between HPV DNA positivity and histological findings of HPV infection was observed.

Although less sensitive than some other molecular biology techniques, in situ hybridization with biotinylated DNA probes proved to be simple and useful for detecting and typing HPV in samples routinely received for histopathological analysis.

Key words: Human papillomavirus; In situ hybridization; Cervical dysplasia; Cervical cancer; Condylomata acuminata.

RESUMO

Amostras de displasias ou carcinomas do colo uterino e de condilomas acuminados da região genital foram analisados retrospectivamente por hibridização in situ (HIS) com sondas de DNA biotiniladas de papilomavirus humano (HPV) tipos 6, 11, 16 e 18. Nenhum caso do grupo controle foi positivo para DNA de HPV. Em displasias leve/moderada, 4 casos (14%) foram positivos para HPV 6 ou 11 e 2 casos (7%), para HPV 16. No grupo de displasia acentuada/carcinoma in situ, 9 casos (31%) tinham DNA de HPV tipos 16 ou 18. Seis carcinomas invasores (20%) foram positivos para HPV tipos 16 ou 18. Entre os condilomas acuminados, 22 casos (73%) foram positivos para HPV tipos 6 ou 11.

Em todos os casos positivos pela HIS somente um tipo viral foi encontrado. Não foi observada correlação entre a positividade para DNA de HPV e achados histológicos de infecção por HPV.

Apesar de menos sensível que algumas outras técnicas de biologia molecular, a hibridização in situ com sondas de DNA biotiniladas mostrou-se simples e adequada para detecção e tipagem de HPV em amostras enviadas rotineiramente para estudo histopatológico.

Full text available only in PDF format.

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Recebido para publicação em 03/12/1991

Aceito para publicação em 13/02/1992

1. BRIGATI, D.J.; MYERSON, D.; LEARY, J.J.; SPALHOLZ, B.; TRAVIS, S.Z.; FONG, C.K.Y.; HSIUNG, C.D. & WARD, D.C. - Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes. Virology, 126: 32-50, 1983.
2. CAUSSY, D.; ORR, W.; DAYA, A.D.; ROTH, P.; REEVES, W. & RAWLS, W. - Evaluation of methods for detecting human papillomavirus DNA sequences in clinical specimens. J. clin. Microbiol. 26: 236-246, 1988.
3. DUGGAN, M.A.; BORAS, V.F.; INOUE, M.; MCGREGOR, S.E. & ROBERTSON, D.I. - Human papillomavirus DNA determination of anal condylomata, dysplasias, and squamous carcinomas with in situ hybridization. Amer. J. clin. Path., 92: 16-21, 1989.
4. FEINBERG, A.P. & VOGELSTEIN, B. - A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analyt. Biochem., 132: 6-13, 1983.
5. FENOGLIO-PREISER, CM. & WILLMAN, C.L. - Molecular biology and the pathologist. Arch. Path. Lab. Med., 111 : 601-619, 1987.
6. FUCHS, P.G.; GIRARDI, F. & PFISTER, H. - Human papillomavirus DNA in normal metaplastic, preneoplastic and neoplastic epithelia of the cervix uteri. Int. J. Cancer, 41: 41-45, 1988.
7. GARUTI, G.; BOSELI, F.; GENAZZANI, A.R.; SILVESTRI, S. & RATTI, G. - Detection and typing of human papillomavirus in histologic specimens by in situ hybridization with biotinylated DNA probes. Amer. J. clin. Path., 92: 604-612, 1989.
8. GRAHAM, A.K.; HERRINGTON, C.S. & McGEE, J.O.D. - Simultaneous in situ genotyping and phenotyping of human papillomavirus cervical lesions: Comparative sensitivity and specificity. J. clin. Path., 44: 96-101, 1991.
9. KULSKI, J.K.; HOWARD, M.J. & PIXLEY, E.C. - DNA sequence of human papillomavirus type 11, 16 and 18 in invasor cervical carcinoma of Western Australian Women. Immunol. Cell. Biol., 65: 77-84, 1987.
10. KURMAN, R.J.; SHIFFMAN, M.H.; LANCASTER, W.D.; REID, R.; JENSON, A.B.; TEMPLE, G.F. & LORINCZ, A.T. - Analysis of individual HPV types in cervical neoplasia. Amer. J. Obstet. Gynec., 159: 293-296, 1988.
11. LANDEGREN, U.; KAISER, R.; CASKEY, C.T. & HOOD, L. - DNA diagnostics - Molecular techniques and automation. Science, 242: 229-237, 1988.
12. LORINCZ, A.T.; TEMPLE, G.F.; KURMAN, R.J.; JENSON, A.B. & LANCASTER, W.D. - Oncogenic association of specific human papillomavirus types with cervical neoplasia. J. nat. Cancer Inst., 79: 671-677, 1987.
13. MCCANCE, D.J.; CAMPION, M.J.; CLARKSON, P.K.; CHESTER, P.M.; JENKINS, D. & SINGER, A. - Prevalence of human papillomavirus type 16 DNA sequences in cervical intraepithelial neoplasia and invasive carcinoma of cervix. Brit. J. Obstet. Gynaec., 92: 1101-1105, 1985.
14. MCCANCE, D.J.; KALACHE, A.; ASHDOWN, K.; ANDRADE, L.; MENEZES, F.; SMITH, P. & DOLL, R. - Human papillomavirus types 16 and 18 in carcinomas of the penis from Brazil. Int. J. Cancer, 37: 55-59, 1986.
15. MEISELS, A.; FORTIN, R. & ROY, M. - Condylomatous lesions of the cervix. II. Cytologic, colposcopic and histopathologic study. Acta Cytol., 21: 379-390, 1977.
16. MIFFLIN, T.E. - Use and applications of nucleic acid probes in the clinical laboratory. Clin. Chem., 35: 1819-1825, 1989.
17. NORVAL, M. & BINGHAM, R.W. - Advances in the use of nucleic acid probes in diagnosis of viral diseases of man. Arch. Virol., 97: 151-165, 1987.
18. NUOVO, G.J. & RICHART, R.M. - A comparison of slot blot, Southern blot, and in situ hybridization analyses for human papillomavirus DNA in genital tract lesions. Obstet. Gynec., 74: 673-678, 1989a.
19. NUOVO, G.J. & RICHART, R.M. - Buffered formalin is the superior fixative for the detection of HPV DNA by in situ hybridization analysis. Amer. J. Path., 134: 837-842, 1989b.
20. NUOVO, G.J.; DARFLER, M.M.; IMPRAIM, C.C. & BROMLEY, S.E. - Occurence of multiple type of human papillomavirus in genital tract lesions. Analysis by in situ hybridization and the polymerase chain reaction. Amer. J. Path., 138: 53-58, 1991.
²¹
ORGANIZACION MUNDIAL DE LA SALUD. - Tipos histológicos de tumores del tracto genital feminino. Ginebra, 1976. 92p. (Clasificación Histologica Internacional de Tumores, 13).
22. OSTROW, R.S.; MANIAS, D.A.; CLARK, B.A.; OKAGAKI, T.; TWIGGS, L.B. & FARAS, A.J. - Detection of human papillomavirus DNA in invasive carcinomas of the cervix by in situ hybridization. Cancer Res., 47: 649-653, 1987.
23. PUROLA, E. & SAVIA, E. - Cytology of gynecologic condyloma acuminatum. Acta. Cytol., 21: 26-31, 1977.
24. PFISTER, H. - Human papillomaviruses and genital cancer. Advanc. Cancer Res., 48: 113-147, 1987.
25. REID, R.; GREENBERG, M.;JENSON, A.B.; HUSAIN, M.; WILLET, J.; DAOUD, Y.; TEMPLE, G. & LORINCE, A.T. - Sexually transmitted papillomaviral infection. I. The anatomic distribution and pathologic grade of neoplastic lesions associated with different viral types. Amer. J. Obstet. Gynec., 156: 212-223, 1987.
26. RICHART, R.M. & NUOVO, G.J. - Human papillomavirus DNA in situ hybridization may be used for the quality control of genital tract biopsies. Obstet. Gynec, 75: 223-226, 1990.
27. SAMBROOK, J.; FRITSCH, E.F. & MANIATIS, T. - Molecular cloning: a laboratory manual. 2Ş ed. New York, Cold Spring Harbor Laboratory Press, 1989.
28. TASE, T.; OKAGAKI, T.; CLARK, B.A.; MANIAS, D.A.; OSTROW, R.S.; TWIGGS, L.B. & FARAS, A.J. - Human papillomavirus types and localization in adenocarcinoma and adenosquamous carcinoma of the uterine cervix: a study by in situ DNA hybridization. Cancer Res., 48: 993-998, 1988.
29. VILLA, L.L. & LOPES, A. - Human papillomavirus DNA sequences in penile carcinomas in Brazil. Int. J. Cancer, 37: 853-855, 1986.
30. VILLA, L.L. & FRANCO, E.L.F. - Epidemiologic correlates of cervical neoplasia and risk of human papillomavirus infection in asymptomatic women in Brazil. J. nat. Cancer Inst., 81: 332-340, 1989.

Address for correspondence:

Prof. Geraldo Brasileiro Filho

Departamento de Anatomia Patológica

Faculdade de Medicina UFMG

Caixa Postal, 340

30.130, Belo Horizonte-MG, Brasil

Publication Dates

Publication in this collection
12 Sept 2006
Date of issue
Aug 1992

History

Received
03 Dec 1991
Accepted
13 Feb 1992

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. BRIGATI, D.J.; MYERSON, D.; LEARY, J.J.; SPALHOLZ, B.; TRAVIS, S.Z.; FONG, C.K.Y.; HSIUNG, C.D. & WARD, D.C. - Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes. Virology, 126: 32-50, 1983.

[2] 2. CAUSSY, D.; ORR, W.; DAYA, A.D.; ROTH, P.; REEVES, W. & RAWLS, W. - Evaluation of methods for detecting human papillomavirus DNA sequences in clinical specimens. J. clin. Microbiol. 26: 236-246, 1988.

[3] 3. DUGGAN, M.A.; BORAS, V.F.; INOUE, M.; MCGREGOR, S.E. & ROBERTSON, D.I. - Human papillomavirus DNA determination of anal condylomata, dysplasias, and squamous carcinomas with in situ hybridization. Amer. J. clin. Path., 92: 16-21, 1989.

[4] 4. FEINBERG, A.P. & VOGELSTEIN, B. - A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analyt. Biochem., 132: 6-13, 1983.

[5] 5. FENOGLIO-PREISER, CM. & WILLMAN, C.L. - Molecular biology and the pathologist. Arch. Path. Lab. Med., 111 : 601-619, 1987.

[6] 6. FUCHS, P.G.; GIRARDI, F. & PFISTER, H. - Human papillomavirus DNA in normal metaplastic, preneoplastic and neoplastic epithelia of the cervix uteri. Int. J. Cancer, 41: 41-45, 1988.

[7] 7. GARUTI, G.; BOSELI, F.; GENAZZANI, A.R.; SILVESTRI, S. & RATTI, G. - Detection and typing of human papillomavirus in histologic specimens by in situ hybridization with biotinylated DNA probes. Amer. J. clin. Path., 92: 604-612, 1989.

[8] 8. GRAHAM, A.K.; HERRINGTON, C.S. & McGEE, J.O.D. - Simultaneous in situ genotyping and phenotyping of human papillomavirus cervical lesions: Comparative sensitivity and specificity. J. clin. Path., 44: 96-101, 1991.

[9] 9. KULSKI, J.K.; HOWARD, M.J. & PIXLEY, E.C. - DNA sequence of human papillomavirus type 11, 16 and 18 in invasor cervical carcinoma of Western Australian Women. Immunol. Cell. Biol., 65: 77-84, 1987.

[10] 10. KURMAN, R.J.; SHIFFMAN, M.H.; LANCASTER, W.D.; REID, R.; JENSON, A.B.; TEMPLE, G.F. & LORINCZ, A.T. - Analysis of individual HPV types in cervical neoplasia. Amer. J. Obstet. Gynec., 159: 293-296, 1988.

[11] 11. LANDEGREN, U.; KAISER, R.; CASKEY, C.T. & HOOD, L. - DNA diagnostics - Molecular techniques and automation. Science, 242: 229-237, 1988.

[12] 12. LORINCZ, A.T.; TEMPLE, G.F.; KURMAN, R.J.; JENSON, A.B. & LANCASTER, W.D. - Oncogenic association of specific human papillomavirus types with cervical neoplasia. J. nat. Cancer Inst., 79: 671-677, 1987.

[13] 13. MCCANCE, D.J.; CAMPION, M.J.; CLARKSON, P.K.; CHESTER, P.M.; JENKINS, D. & SINGER, A. - Prevalence of human papillomavirus type 16 DNA sequences in cervical intraepithelial neoplasia and invasive carcinoma of cervix. Brit. J. Obstet. Gynaec., 92: 1101-1105, 1985.

[14] 14. MCCANCE, D.J.; KALACHE, A.; ASHDOWN, K.; ANDRADE, L.; MENEZES, F.; SMITH, P. & DOLL, R. - Human papillomavirus types 16 and 18 in carcinomas of the penis from Brazil. Int. J. Cancer, 37: 55-59, 1986.

[15] 15. MEISELS, A.; FORTIN, R. & ROY, M. - Condylomatous lesions of the cervix. II. Cytologic, colposcopic and histopathologic study. Acta Cytol., 21: 379-390, 1977.

[16] 16. MIFFLIN, T.E. - Use and applications of nucleic acid probes in the clinical laboratory. Clin. Chem., 35: 1819-1825, 1989.

[17] 17. NORVAL, M. & BINGHAM, R.W. - Advances in the use of nucleic acid probes in diagnosis of viral diseases of man. Arch. Virol., 97: 151-165, 1987.

[18] 18. NUOVO, G.J. & RICHART, R.M. - A comparison of slot blot, Southern blot, and in situ hybridization analyses for human papillomavirus DNA in genital tract lesions. Obstet. Gynec., 74: 673-678, 1989a.

[19] 19. NUOVO, G.J. & RICHART, R.M. - Buffered formalin is the superior fixative for the detection of HPV DNA by in situ hybridization analysis. Amer. J. Path., 134: 837-842, 1989b.

[20] 20. NUOVO, G.J.; DARFLER, M.M.; IMPRAIM, C.C. & BROMLEY, S.E. - Occurence of multiple type of human papillomavirus in genital tract lesions. Analysis by in situ hybridization and the polymerase chain reaction. Amer. J. Path., 138: 53-58, 1991.

[21] ²¹
ORGANIZACION MUNDIAL DE LA SALUD. - Tipos histológicos de tumores del tracto genital feminino. Ginebra, 1976. 92p. (Clasificación Histologica Internacional de Tumores, 13).

[22] 22. OSTROW, R.S.; MANIAS, D.A.; CLARK, B.A.; OKAGAKI, T.; TWIGGS, L.B. & FARAS, A.J. - Detection of human papillomavirus DNA in invasive carcinomas of the cervix by in situ hybridization. Cancer Res., 47: 649-653, 1987.

[23] 23. PUROLA, E. & SAVIA, E. - Cytology of gynecologic condyloma acuminatum. Acta. Cytol., 21: 26-31, 1977.

[24] 24. PFISTER, H. - Human papillomaviruses and genital cancer. Advanc. Cancer Res., 48: 113-147, 1987.

[25] 25. REID, R.; GREENBERG, M.;JENSON, A.B.; HUSAIN, M.; WILLET, J.; DAOUD, Y.; TEMPLE, G. & LORINCE, A.T. - Sexually transmitted papillomaviral infection. I. The anatomic distribution and pathologic grade of neoplastic lesions associated with different viral types. Amer. J. Obstet. Gynec., 156: 212-223, 1987.

[26] 26. RICHART, R.M. & NUOVO, G.J. - Human papillomavirus DNA in situ hybridization may be used for the quality control of genital tract biopsies. Obstet. Gynec, 75: 223-226, 1990.

[27] 27. SAMBROOK, J.; FRITSCH, E.F. & MANIATIS, T. - Molecular cloning: a laboratory manual. 2Ş ed. New York, Cold Spring Harbor Laboratory Press, 1989.

[28] 28. TASE, T.; OKAGAKI, T.; CLARK, B.A.; MANIAS, D.A.; OSTROW, R.S.; TWIGGS, L.B. & FARAS, A.J. - Human papillomavirus types and localization in adenocarcinoma and adenosquamous carcinoma of the uterine cervix: a study by in situ DNA hybridization. Cancer Res., 48: 993-998, 1988.

[29] 29. VILLA, L.L. & LOPES, A. - Human papillomavirus DNA sequences in penile carcinomas in Brazil. Int. J. Cancer, 37: 853-855, 1986.

[30] 30. VILLA, L.L. & FRANCO, E.L.F. - Epidemiologic correlates of cervical neoplasia and risk of human papillomavirus infection in asymptomatic women in Brazil. J. nat. Cancer Inst., 81: 332-340, 1989.

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