Fonseca Renata Fragelli, Rosas Siane Lopes Bittencourt, Oliveira José Antônio, Teixeira Anselmo, Alves Gilda, Carvalho Maria da Glória Costa
Congenital Malformations Laboratory, Department of Genetics, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
Molecular Oncology Laboratory, Clementino Fraga Filho University Hospital.
Sao Paulo Med J. 2015 Feb;133(1):51-4. doi: 10.1590/1516-3180.2013.1912814. Epub 2015 Jan 1.
The Epstein-Barr virus (EBV) is the most common cause of infectious mononucleosis and is also associated with several human tumors, including Burkitt's lymphoma, Hodgkin's lymphoma, some cases of gastric carcinoma and nasopharyngeal carcinoma, among other neoplasms. The aim of this study was to screen 75 primary gliomas for the presence of specific EBV DNA sequences by means of the polymerase chain reaction (PCR), with confirmation by direct sequencing.
Prevalence study on EBV molecular genetics at a molecular pathology laboratory in a university hospital and at an applied genetics laboratory in a national institution.
A total of 75 primary glioma biopsies and 6 others from other tumors from the central nervous system were obtained. The tissues were immediately frozen for subsequent DNA extraction by means of traditional methods using proteinase K digestion and extraction with a phenol-chloroform-isoamyl alcohol mixture. DNA was precipitated with ethanol, resuspended in buffer and stored. The PCRs were carried out using primers for amplification of the EBV BamM region. Positive and negative controls were added to each reaction. The PCR products were used for direct sequencing for confirmation.
The viral sequences were positive in 11/75 (14.7%) of our samples.
The prevalence of EBV DNA was 11/75 (14.7%) in our glioma collection. Further molecular and epidemiological studies are needed to establish the possible role played by EBV in the tumorigenesis of gliomas.
爱泼斯坦-巴尔病毒(EBV)是传染性单核细胞增多症最常见的病因,还与多种人类肿瘤相关,包括伯基特淋巴瘤、霍奇金淋巴瘤、部分胃癌和鼻咽癌等其他肿瘤。本研究旨在通过聚合酶链反应(PCR)筛查75例原发性胶质瘤中特定EBV DNA序列的存在情况,并通过直接测序进行确认。
在一所大学医院的分子病理学实验室和一家国家机构的应用遗传学实验室进行的关于EBV分子遗传学的患病率研究。
共获取75例原发性胶质瘤活检样本以及6例来自中枢神经系统其他肿瘤的样本。组织立即冷冻,随后采用传统方法,用蛋白酶K消化并用苯酚-氯仿-异戊醇混合物提取DNA。DNA用乙醇沉淀,重悬于缓冲液中并储存。使用扩增EBV BamM区域的引物进行PCR。每个反应均添加阳性和阴性对照。PCR产物用于直接测序以进行确认。
我们的样本中有11/75(14.7%)的病毒序列呈阳性。
在我们收集的胶质瘤样本中,EBV DNA的患病率为11/75(14.7%)。需要进一步的分子和流行病学研究来确定EBV在胶质瘤肿瘤发生中可能发挥的作用。