Epstein-Barr virus (EBV) has a pathogenic role in several lymphomas including diffuse large B-cell lymphoma (DLBCL). In this study, we detected EBV from formalin-fixed paraffin embedded (FFPE) tissues of DLBCL patients by RT-PCR and compared the sensitivity of the RT-PCR method to in situ hybridizat...
Epstein-Barr virus (EBV) has a pathogenic role in several lymphomas including diffuse large B-cell lymphoma (DLBCL). In this study, we detected EBV from formalin-fixed paraffin embedded (FFPE) tissues of DLBCL patients by RT-PCR and compared the sensitivity of the RT-PCR method to in situ hybridization (ISH) method. The RNA was extracted from 91 FFPE samples with DLBCL and amplified with primers targeting EBV-encoded small RNA (EBER) by RT-PCR. When using the RT-PCR method, 13 of 91 patients (14.3%) were positive and among these 13 cases, 7 cases (7.7%) were from > 50-year-old patients that is classified as EBV positive DLBCL of the elderly. In previous results using ISH method, 3 of 91 patients (3.3%) were positive and 2 case (2.4%) were older than 50-year-old. These results indicate that RT-PCR method used in this study shows a higher sensitivity than ISH method. The ratio of male versus female among the EBV positive samples was 1.2:1 with the ratio of male higher. If RT-PCR method having high sensitivity is used simultaneously as well as the ISH method providing the information of the EBV positive cellular location, it is expected that EBV will be more accurately detected.
Epstein-Barr virus (EBV) has a pathogenic role in several lymphomas including diffuse large B-cell lymphoma (DLBCL). In this study, we detected EBV from formalin-fixed paraffin embedded (FFPE) tissues of DLBCL patients by RT-PCR and compared the sensitivity of the RT-PCR method to in situ hybridization (ISH) method. The RNA was extracted from 91 FFPE samples with DLBCL and amplified with primers targeting EBV-encoded small RNA (EBER) by RT-PCR. When using the RT-PCR method, 13 of 91 patients (14.3%) were positive and among these 13 cases, 7 cases (7.7%) were from > 50-year-old patients that is classified as EBV positive DLBCL of the elderly. In previous results using ISH method, 3 of 91 patients (3.3%) were positive and 2 case (2.4%) were older than 50-year-old. These results indicate that RT-PCR method used in this study shows a higher sensitivity than ISH method. The ratio of male versus female among the EBV positive samples was 1.2:1 with the ratio of male higher. If RT-PCR method having high sensitivity is used simultaneously as well as the ISH method providing the information of the EBV positive cellular location, it is expected that EBV will be more accurately detected.
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제안 방법
, 2007). In this study, we used RT-PCR method and targeted EBER to detect EBV infection from the FFPE samples of the DLBCL patients.
The size of the product was compared to 100 bp DNA ladder (Fermentas, Burlington, Canada). The sequences of the primers are as follows: GAT CCA AAC TTT AGT TTT AG (EBER1 forward), GCG AAC CGT AAC TCT ATA C (EBER1 reverse), CCA CAT GGC TCA GAC ACC AT (GAPDH 71 bp forward), ACC AGG CGC CCA ATA CG (GAPDH 71 bp reverse), CCA CAT GGC TCA GAC ACC AT (GAPDH 153 bp forward), GTA AAC CAT GTA GTT GAG GTC (GAPDH 153 bp reverse), TAT GAC AAC AGC CTC AAG AT (GAPDH 316 bp forward), and AGG TCC ACC ACT GAC ACG TT (GAPDH 316 bp reverse). The primers for EBER1 were designed in our study group and specificity of the primers was confirmed by sequencing of the amplicon in our recent report (Lee et al.
이론/모형
In this study, we extracted RNA from FFPE tissue samples of DLBCL patients and detected expression of EBER of EBV by using RT-PCR method. Here we found that RT- PCR method with the primer we used could more sensitively detect EBV than ISH method which is a gold standard method for EBV detection.
참고문헌 (31)
Anagnostopoulos I, Hummel M. Epstein-Barr virus in tumours. Histopathology. 1996. 29: 297-315.
Beltran BE, Castillo JJ, Morales D, de Mendoza FH, Quinones P, Miranda RN, Gallo A, Lopez-Ilasaca M, Butera JN, Sotomayor EM. EBV-positive diffuse large B-cell lymphoma of the elderly: a case series from Peru. Am J Hematol. 2011. 86: 663-667.
Chang KL, Chen YY, Shibata D, Weiss LM. Description of an in situ hybridization methodology for detection of Epstein-Barr virus RNA in paraffin-embedded tissues, with a survey of normal and neoplastic tissues. Diagn Mol Pathol. 1992. 1: 246-255.
Epstein A. Thirty years of Epstein-Barr virus. Epstein-Barr virus Report. 1994. 1: 3-4.
Gibson SE, Hsi ED. Epstein-Barr virus-positive B-cell lymphoma of the elderly at a United States tertiary medical center: an uncommon aggressive lymphoma with a nongerminal center B-cell phenotype. Hum Pathol. 2009. 40: 653-661.
Hoeller S, Tzankov A, Pileri SA, Went P, Dirnhofer S. Epstein-Barr virus-positive diffuse large B-cell lymphoma in elderly patients is rare in Western populations. Hum Pathol. 2010. 41: 352-357.
Hofscheier A, Ponciano A, Bonzheim I, Adam P, Lome-Maldonado C, Vela T, Cortes E, Ortiz-Hidalgo C, Fend F, Quintanilla-Martinez L. Geographic variation in the prevalence of Epstein-Barr virus-positive diffuse large B-cell lymphoma of the elderly: a comparative analysis of a Mexican and a German population. Mod Pathol. 2011. 24: 1046-1054.
Iwakiri D, Eizuru Y, Tokunaga M, Takada K. Autocrine growth of Epstein-Barr virus-positive gastric carcinoma cells mediated by an Epstein-Barr virus-encoded small RNA. Cancer Res. 2003. 63: 7062-7067.
Kashofer K, Viertler C, Pichler M, Zatloukal K. Quality control of RNA preservation and extraction from paraffin-embedded tissue: implications for RT-PCR and microarray analysis. PLoS One. 2013. 8: e70714.
Kitagawa N, Goto M, Kurozumi K, Maruo S, Fukayama M, Naoe T, Yasukawa M, Hino K, Suzuki T, Todo S, Takada K. Epstein-Barr virus-encoded poly(A)(-) RNA supports Burkitt's lymphoma growth through interleukin-10 induction. EMBO J. 2000. 19: 6742-6750.
Kuze T, Nakamura N, Hashimoto Y, Sasaki Y, Abe M. The characteristics of Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma: comparison between EBV(+) and EBV(-) cases in Japanese population. Jpn J Cancer Res. 2000. 91: 1233-1240.
Lee J, Park M, Lee MH, Woo HJ, Kim HW, Yang JY, Eom YB, Kim SH, Yoon C, Kim JB. Development of EBV-encoded small RNA targeted PCR to classify EBV positive diffuse large B-cell lymphoma (DLBCL) of the elderly. Int J Clin Exp Pathol. 2015. 8: 7859-7868.
Montes-Moreno S, Odqvist L, Diaz-Perez JA, Lopez AB, de Villambrosia SG, Mazorra F, Castillo ME, Lopez M, Pajares R, Garcia JF, Mollejo M, Camacho FI, Ruiz-Marcellan C, Adrados M, Ortiz N, Franco R, Ortiz-Hidalgo C, Suarez-Gauthier A, Young KH, Piris MA. EBV-positive diffuse large B-cell lymphoma of the elderly is an aggressive post-germinal center B-cell neoplasm characterized by prominent nuclear factor- ${\kappa}B$ activation. Mod Pathol. 2012. 25: 968-982.
Nanbo A, Inoue K, Adachi-Takasawa K, Takada K. Epstein-Barr virus RNA confers resistance to interferon-alpha-induced apoptosis in Burkitt's lymphoma. EMBO J. 2002. 21: 954-965.
Oyama T, Yamamoto K, Asano N, Oshiro A, Suzuki R, Kagami Y, Morishima Y, Takeuchi K, Izumo T, Mori S, Ohshima K, Suzumiya J, Nakamura N, Abe M, Ichimura K, Sato Y, Yoshino T, Naoe T, Shimoyama Y, Kamiya Y, Kinoshita T, Nakamura S. Age-related EBV-associated B-cell lymphoproliferative disorders constitute a distinct clinicopathologic group: a study of 96 patients. Clin Cancer Res. 2007. 13: 5124-5132.
Park S, Lee J, Ko YH, Han A, Jun HJ, Lee SC, Hwang IG, Park YH, Ahn JS, Jung CW, Kim K, Ahn YC, Kang WK, Park K, Kim WS. The impact of Epstein-Barr virus status on clinical outcome in diffuse large B-cell lymphoma. Blood. 2007. 110: 972-978.
Qi ZL, Han XQ, Hu J, Wang GH, Gao JW, Wang X, Liang DY. Comparison of three methods for the detection of Epstein-Barr virus in Hodgkin's lymphoma in paraffin-embedded tissues. Mol Med Rep. 2013. 7: 89-92.
Rosa MD, Gottlieb E, Lerner MR, Steitz JA. Striking similarities are exhibited by two small Epstein-Barr virus-encoded ribonucleic acids and the adenovirus-associated ribonucleic acids VAI and VAII. Mol Cell Biol. 1981. 1: 785-796.
Smith A, Howell D, Patmore R, Jack A, Roman E. Incidence of haematological malignancy by sub-type: a report from the Haematological Malignancy Research Network. Br J Cancer. 2011. 105: 1684-1692.
Smith A, Roman E, Howell D, Jones R, Patmore R, Jack A. The Haematological Malignancy Research Network (HMRN): a new information strategy for population based epidemiology and health service research. Br J Haematol. 2010. 148: 739-753.
The Non-Hodgkin's Lymphoma Classification Project. A clinical evaluation of the International Lymphoma Study Group classification of non-Hodgkin's lymphoma. Blood. 1997. 89: 3909-3918.
Vinay K, Abul KA, Nelson F, Jon CA. Robbins & Cotran Pathologic Basis of Disease. 2010. 607. Saunders. Philadelphia.
Yang L, Aozasa K, Oshimi K, Takada K. Epstein-Barr virus (EBV)-encoded RNA promotes growth of EBV-infected T cells through interleukin-9 induction. Cancer Res. 2004. 64: 5332-5337.
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