African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5211

Full Length Research Paper

In situ hybridization and polymerase chain reaction methods for the detection of Epstein-Barr virus RNA in breast cancer specimens

Izham N. S. A. S.1, H. Zuridah1, 2*, S. Rashidah1, O. Azizon3 and N. A. Zeenathul4
  1Faculty of Health Sciences, Universiti Teknologi MARA, 42300 Puncak Alam, Selangor, Malaysia. 2Pharmogenomic Centre (PROMISE), Faculty of Pharmacy, Universiti Teknologi MARA, 42300 Puncak Alam, Selangor, Malaysia. 3Pathology Department, Hospital Tuanku Ja’afar, Jalan Rasah, 70300 Seremban, NegeriSembilan, Malaysia. 4Department of Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Malaysia.
Email: [email protected]

  •  Accepted: 08 September 2011
  •  Published: 09 November 2011



The role of Epstein-Barr virus (EBV) as a cofactor in breast cancer is controversial and its association with breast cancer varies. In this study, EBV was detected by using in situhybridization technique (ISH) to detect Epstein-barr virus encoded RNA1 (EBER1) transcripts.  Archival formalin-fixed paraffin embedded breast cancer tissue samples (n = 139) and normal breast tissue (n = 20) obtained from Hospital Tuanku Ja’afar were sectioned, stained and examined microscopically for nuclear staining and by DNA amplification of the same gene. By ISH, 83/139 (59.7%) and 12/20 (60%) were EBV positives in the breast cancer tissues and normal tissues, respectively. On the other hand, confirmation by polymerase chain reaction (PCR) found that additional six samples (89/139 or 64%) breast cancer tissues were positive for EBER1 gene. To further confirm the identity of these amplified products, two samples (UiTM-53 and UiTM-73) were sequenced, BLAST, analyzed phylogenetically and was found to be 100% similar to the EBV EBER1 gene sequences already deposited in the GenBank (accession numbers AB065135, FN545286, EF187853 and DQ883818). These preliminary findings suggest that there is a correlation between EBV and breast cancer but need further testing with more samples to confirm the role of EBV.


Key words: Breast cancer, Epstein-Barr virus, EBER1 transcripts, in situ hybridization, polymerase chain reaction (PCR), tumour grading.