Lack of Human Cytomegalovirus in Gliomas. A Viral Load Analysis by Quantitative Real Time–PCR
Araceli Garcia-Martinez, Esperanza Irles, Victor M Barbera, Cecilia Egoavil, Cristina Alenda, Adela Castillejo, Maria Isabel Castillejo, Enrique Ochoa, Mari Carmen Barea, Teresa Quintanar, Laura Heredia, Pedro Moreno, Joaquin Herrero, Maria Jose Portugues, Ana Culianez, Encarna Andrada, Javier Sendra, Alvaro Rodriguez-Lescure, Artemio Paya, Ignacio Aranda, Jose Luis Soto. Elche University Hospital, Elche, Alicante, Spain; Alicante University Hospital, Alicante, Spain; Castellon Provincial Hospital, Castellon, Spain
Background: Recently, a consensus has been reached that there is sufficient evidence to conclude that human cytomegalovirus (HCMV) sequences and viral gene expression exist in most, if not all, malignant gliomas. However, no previous report has approached a quantitative analysis of the HCMV in gliomas. We aimed to study the HCMV viral load in different stages of human primary gliomas.
Design: A total of 82 paraffin embedded tissues of human primary gliomas (stages II: 15; III: 13 and IV: 54) from the Elche & Alicante Hospital Biobanks were included in this study. DNA was isolated using a high performance forensic kit (QIAamp DNA Investigator; Qiagen). HCMV was analyzed by Real Time PCR using a standardized kit for HCMV quantitation in diagnostic (RealStar CMV PCR Kit 1.0; Altona DIAGNOSTICS). Positive and negative controls were included. Each DNA sample was tested in triplicate.
Results: A six points standard curve (Pearson's correlation coefficient >0.99) was used to interpolate the HCMV viral load from 10 to 10.000 copies, unexpectedly 79 samples of this serie of gliomas showed undetectable DNA of HCMV. Three cases showed a low number of viral copies (<10) (stages III: 2; IV: 1). The strength of the analytical accuracy components required to characterize a quantitative test (trueness, precision and limit of detection) is assured by the employed methodology. The controversy regarding the existence and role of HCMV in gliomas has been debated in the literature. The recent consensus pointing to an important role of HCMV in gliomas needs to be reviewed.
Conclusions: Our results strongly support that HCMV is nearly absent in gliomas. The role of HCMV infection in glioma development is irrelevant.
Wednesday, March 6, 2013 1:00 PM
Poster Session VI # 243, Wednesday Afternoon