Acta Medica Iranica 2018. 56(7):434-440.

Detection and Genotyping of Epstein-Bar Virus Among Paraffin Embedded Tissues of Hodgkin and Non-Hodgkin's Lymphoma Patients in Ahvaz, Iran
Ala Habibian, Manoochehr Makvandi, Alireza Samarbaf-Zadeh, Niloofar Neisi, Rahim Soleimani-Jelodar, Kamyar Makvandi, Shima Izadi


Epstein-Barr virus (EBV) is a DNA virus which belongs to the Herpesviridae family, has been infected with 90% of the world’s population. EBV is transmitted through saliva and associated with different diseases such as Infectious Mononucleosis, Nasopharyngeal Carcinoma, Burkitt`s lymphoma, Hodgkin and Non-Hodgkin`s lymphoma. This study was designed to determine the frequency of EBV DNA and genotyping of this virus in histological tissues of Hodgkin and Non-Hodgkin's lymphoma in Ahvaz, Iran. In this study, 12 samples of Hodgkin and 29 samples of Non-Hodgkin`s lymphoma were examined from Ahvaz Imam Khomeini Hospital. After deparaffinization and DNA extraction, Nested-PCR technique was carried out on Epstein-Barr nuclear antigen 1 (EBNA-1) and Epstein-Barr nuclear antigen 3C (EBNA-3C) regions in order to detect EBV genome and genotype in tumoral tissues. Among 12 cases of Hodgkin`s lymphoma, 3 (25%) cases and among 29 cases of Non-Hodgkin's lymphoma, 14 (48%) cases were positive for EBV. All 3 EBV positive cases of Hodgkin`s lymphoma and a number of 8 (57%) cases of Non-Hodgkin`s lymphoma belonged to the adolescence age group which proves an association between age and EBV positive in Hodgkin and Non-Hodgkin`s lymphoma according to Fisher`s exact test (P=0.03). All EBV positive cases in Hodgkin and Non-Hodgkin`s lymphoma were EBV-1. The results of this study revealed that EBV played a possible important role in Hodgkin and Non-Hodgkin`s lymphoma especially in adults. Also, the main genotype of virus in the current study was EBV-1.


EBNA-1; EBNA-3C; Epstein-Barr virus; Hodgkin lymphoma; Non-hodgkin lymphoma

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Arvin A, Campadelli G, Mocarski E, Moore PRB, Whitley R, Yamanishi K. Human Herpes Virus Arvin A, Campadelli G, Mocarski E, Moore PRB, Whitley R, Yamanishi K, editors. New York: Cambridge University ; 2007.

Brink A, Vervoort M, Middeldorp J, Meijer C, van den Brule A. Nucleic Acid Sequence-Based Amplification, a New Method for Analysis of Spliced and Unspliced Epstein-Barr Virus Latent Transcripts, and Its Comparison with Reverse Transcriptase PCR. J. Clin. Microbiol. 1998; 36(11): 3164-3169.

Fields BN, Knipe DM, Howley PM. Fields Virology. 6th ed. Cohen JI, Griffin DE, Lamb RA, Martin MA, Racaniello VR, Roizman B, editors. Philadelphia: Lippincott Williams & Wilkins; 2013.

Pereira de Lima MA, Pitombeira Ferreira MV, Pessoa Barros MA, Inês de Moura Campos Pardini M, Camargo Ferrasi A, Barem Rabenhorst SH. Epstein-Barr virus-associated gastric carcinoma in Brazil: comparison between in situ hybridization and polymerase chain reaction detection. Braz J Microbiol. 2012 Jan./Mar.; 43(1): 393-404.

Klein G, Klein E, Kashuba E. Interaction of Epstein-Barr virus (EBV) with human B-lymphocytes. Biochemical and Biophysical Research Communications. 2010; 396: 67-73.

Stevens SJ, Vervoort MB, van den Brule A, Meenhorst PL, Meijer C, Middeldorp J. Monitoring of Epstein-Barr Virus DNA Load in Peripheral Blood by Quantitative Competitive PCR. J Clin Microbiol. 1999, 37(9): 2852–7.

Carbone A, Gloghini A, Dotti G. EBV-Associated Lymphoproliferative Disorders: Classification and Treatment. Oncologist. 2008; 13: 577-85.

Ai J, Xie Z, Liu C, Huang Z, Xu J. Analysis of EBNA-1 and LMP-1 variants in diseases associated with EBV infection in Chinese children. Virol J. 2012; 9(13), doi:10.1186/1743-422X-

Hassan R, White LR, Gustavo Stefanoff C, Elgui de Oliveira D, Felisbino F, Esteves Klumb C, et al. Epstein-Barr Virus (EBV) detection and typing by PCR: a contribution to diagnostic screening of EBV-positive Burkitt's lymphoma. Diagn Pathol. 2006; 1(17), doi:10.1186/1746-1596-1-17.

Kim I, Park ER, Park SH, Lin Z, Kim YS. Characteristics of Epstein-Barr Virus Isolated From The Malignant Lymphomas in Korea. J Med Virol. 2002; 67: 59-66.

Ocheni S, Olusina DB, Oyekunle AA, Ibegbulam OG, Kröger N, Bacher U, et al. EBV-Associated Malignancies. The Open Infect Dis J. 2010; 4: 101-112.

Wang WY, Chien YC, Jan JS, Chueh CM, Lin JC. Consistent Sequence Variation of Epstein-Barr Virus Nuclear Antigen 1 in Primary Tumor and Peripheral Blood Cells of Patients with Nasopharyngeal Carcinoma. Clin Cancer Res. 2002 August; 8: 2586–2590.

Skare J, Edson C, Farley J, Strominger J. The B95-8 isolate of Epstein-Barr virus arose from an isolate with a standard genome. J Virol. 1982, December; 44(3): 1088-1091.

Carbone A, Cesarman E, Spina M, Gloghini A, Schulz T. HIV-associated lymphomas and gamma-herpesviruses. Blood. 2009, February 5; 113(6): 1213-1224.

Gulley ML. Molecular Diagnosis of Epstein-Barr Virus-Related Diseases. J Mol Diagn. 2001 February; 3(1).

Tumwine LK, Orem J, Kerchan P, Byarugaba W, Pileri SA. EBV,HHV8 and HIV in B cell non Hodgkin lymphoma in Kampala, Uganda. Infect Agent Cancer. 2010 June 30; 5(12): 1-7, doi: 10.1186/1750-9378-5-12.

Hjalgrim H, Askling J, Madsen M, Rosdahl N, Storm HH. Risk of Hodgkin’s Disease and Other Cancers After Infectious Mononucleosis. J Natl Cancer I. 2000 September; 92(18): 1522-8.

Farrell K, Jarrett RF. The molecular pathogenesis of Hodgkin Lymphoma. Histopathology. 2011 january; 58(1): 15-25.

Čičkušić E, Mustedanagić-Mujanović J, Iljazović E, Karasalihović Z, Škaljić I. Association of Hodgkin’s Lymphoma with Epstein Barr Virus Infection. Bosn J Basic Med Sci. 2007 February; 7(1): 58-65.

Flavell KJ, Murray PG. Hodgkin’s disease and the Epstein-Barr virus. J Clin Pathol. 2000 October; 53(5): 262-9.

Levin LI, Chang ET, Ambinder RF, Lennette ET, Rubertone MV, Mann RB, et al. Atypical prediagnosis Epstein-Barr virus serology restricted to EBV-positive Hodgkin lymphoma. Blood. 2012 September; 120 (18): 3750-5

Glaser SL, Hsu JL. Hodgkin's disease in Asians: Incidence patterns and risk factors in population-based data. Leukemia Res. 2002; 26(3): 261-9.

Shenoy P, Maggioncalda A, Malik N, Flowers C. Incidence Patterns and Outcomes for Hodgkin Lymphoma Patients in the United States. Adv Hematol2011, doi:10.1155/2011/725219.

Macsween KF, Crawford DH. Epstein-Barr virus—recent advances. Lancet Infect Dis. 2003 March; 3: 131-40.

Bakshi N, Maghfoor I. The Current Lymphoma Classification: New Concepts and Practical Applications—Triumphs and Woes. Ann Saudi Med. 2012 May-June; 32(3): 296-305.

Steidl C, Connors JM, Gascoyne RD. Molecular Pathogenesis of Hodgkin’s Lymphoma: Increasing Evidence of the Importance of the Microenvironment. J Clin Oncol. 2011 April 11, doi: 10.1200/JCO.2010.32.8401.

Najafipour S, Mokhtari Azad T, Kousari F, Mahmoodi M, Murray PG, Nategh R. Association of Epstein-Barr virus and hodgkin disease. Acta Medica Iraninca. 2003; 41(1): 1-10.

Al-Salam S, John A, Daoud S, Chong S, Castella A. Expression of Epstein-Barr virus in Hodgkin lymphoma in a population of United Arab Emirates nationals. Leuk Lymphoma. 2008 September; 49(9): 1769-77.

Barros MH, Hassan R, Niedobitek G. Disease patterns in pediatric classical Hodgkin lymphoma: a report from a developing area in Brazil. Hematol Oncol. 2011 December; 29(4): 190-5.

Diepstra A, Van Imhoff GW, Schaapveld M, Karim-Kos H, van den Berg A, Vellenga E, et al. Latent Epstein-Barr virus infection of tumor cells in classical Hodgkin's lymphoma predicts adverse outcome in older adult patients. J Clin Oncol. 2009 August 10; 27(23): 3815-21.

Zhao P, Lu Y, Liu L, Zhong M. Aberrant cytoplasmic expression of cyclin B1 protein and its correlation with EBV-LMP1, P53 and P16(INK4A) in classical Hodgkin lymphoma in China. Pathol Oncol Res. 2011 June; 17(2): 369-73.

Zhou XG, Hamilton-Dutoit SJ, Yan QH, Pallesen G. The association between Epstein-Barr virus and Chinese Hodgkin's disease. Int J Cancer. 1993 September; 55(3): 359-63.

Zarate-Osorno A, Roman LN, Kingma DW, Meneses-Garcia A, Jaffe ES. Hodgkin's disease in Mexico. Prevalence of Epstein-Barr virus sequences and correlations with histologic subtype. Cancer. 1995 March 15; 75(6): 1360-6.

Beiderbeck AB, Holly EA, Sturkenboom MCJM, Coebergh JWW, Stricker BHC, Leufkens HGM. Prescription Medications Associated with a Decreased Risk of Non-Hodgkin’s Lymphoma. Am J Epidemiol. 2003; 157(6): 510-516.

Basiratnia M, Baradaran-Heravi A, Yavarian M, Geramizadeh B, Karimi M. Non-Hodgkin Lymphoma in a Child with Schimke Immuno-Osseous Dysplasia. Iran J Med Sci. 2011 September; 36(3): 222-5.

Fisher SG, Fisher RI. The epidemiology of non-Hodgkin’s lymphoma. Oncogene. 2004; 23: 6524–34.

Evens AM, Winter JN, Gordon LI, Chiu BC, Tsang R, Rosen ST. CancerNetwork home of the journal oncology. [Online].; 2011 [cited 2013 1 24. Available from: lymphoma/article/10165/1802748.

Kasprzak A, Spachacz R, Wachowiak J, Stefañska K, Zabel M. Epstein-Barr virus (EBV) infection in B-cell non-Hodgkin's lymphomas in children: virus latency and its correlation with CD21 and CD23 molecules. Folia Histochem Cyto. 2007; 45(3): 169-179.

Pourakbari B, mamishi S, Pajand O, Naji AR, Mahjoub F, Kouchakzadeh L, et al. Detection of Epstein- Barr virus infection in lymphoma: ELISA and PCR method. Tehran Univ Med J. 2010 February; 67(11): 787-792.

Kosari F, Yarigarravesh HR, Rezvan M. Determining the correlation of Epstein-Barr virus with diffuse large B-cell lymphoma by chromogenic in situ hybridization. Tehran Univ Med J. 2012 September; 70(6): 351-6.

Mitarnun W, Pradutkanchana J, Takao S, Saechan V, Suwiwat S, Ishida T. Epstein-barr virus-associated non-Hodgkin's lymphoma of B-cell origin, Hodgkin's disease, acute leukemia, and systemic lupus erythematosus: a serologic and molecular analysis. J Med Assoc Thai. 2002 May; 85(5): 552-559.

Chabay P, Lara J, Lorenzetti M, Cambra P, Acosta Haab G, Aversa L, et al. Epstein Barr virus in relation to apoptosis markers and patients' outcome in pediatric B-cell non-Hodgkin lymphoma. Cancer Lett. 2011 May 4; 307(2): 221-6.

Bahnassy AA, Zekri ARN, El-Houssini S, Khalid HM, Sedky LM, Mokhtar NM. epstein barr virus in head and neck extranodal non-hodgkin Lymphoma in Egypt. Journal of Egyptian Nat. Cancer Inst. 2003 December; 15(4): 349-62.

Umar CS. New Developments in Epstein-Barr Virus Research. In Umar CS, editor.. New York: Nova Publishers; 2006.

Frappier L. The Epstein-Barr Virus EBNA1 Protein. Scientifica2012: 1-15. Article ID: 438204

Frappier LD. Details about NEW EBNA1 and Epstein-Barr Virus Associated Tumours: Springer publisher, 2013.

Robaina TF, Valladares CP, Tavares DS, Napolitano WC, Silva LE, Dias EP, et al. Polymerase chain reaction genotyping of Epstein-Barr virus in scraping samples of the tongue lateral border in HIV-1 seropositive patients. Mem Inst Oswaldo Cruz, Rio de Janeiro. 2008 June; 103(4): 326-31.


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