Noninvasive diagnosis of nasopharyngeal carcinoma: Nasopharyngeal brushings reveal high Epstein‐Barr virus DNA load and carcinoma‐specific viral BARF1 mRNA

• Published in: International journal of cancer Vol. 119; no. 3; pp. 608 - 614
• Main Authors: Stevens, Servi J.C., Verkuijlen, Sandra A.W.M., Hariwiyanto, Bambang, Harijadi, Paramita, Dewi K., Fachiroh, Jajah, Adham, Marlinda, Tan, I. Bing, Haryana, Sophia M., Middeldorp, Jaap M.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.08.2006; Wiley-Liss
• Subjects: Animals; Biological and medical sciences; carcinoma; Cell Line, Transformed; Cell Line, Tumor; Cytodiagnosis - methods; diagnosis; DNA, Viral - blood; DNA, Viral - genetics; EBV; Ent. Stomatology; Epstein-Barr virus; Epstein-Barr Virus Infections - blood; Epstein-Barr Virus Infections - diagnosis; Epstein-Barr Virus Infections - virology; Epstein-Barr Virus Nuclear Antigens - genetics; Herpesvirus 4, Human - genetics; Humans; Indonesia; Investigative techniques, diagnostic techniques (general aspects); Medical sciences; Nasopharyngeal Neoplasms - blood; Nasopharyngeal Neoplasms - diagnosis; Nasopharyngeal Neoplasms - virology; Nasopharynx - pathology; Nasopharynx - virology; NPC; oncogene; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Prospective Studies; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sensitivity and Specificity; Tumors; Viral Load; Viral Matrix Proteins - genetics; Viral Proteins - genetics; Indonesia; NPC; Nasopharynx cancer; Brushing; Epstein Barr virus; Nasopharynx carcinoma; Cancerology; Messenger RNA; EBV; ENT disease; Pharynx disease; Diagnosis; Viral load; Nucleic acid; Gammaherpesvirinae; Human; Herpesviridae; Malignant tumor; oncogene; Infection; Virus; Amplification; carcinoma; DNA; Viral disease; Non invasive method; Molecular biology
• ISSN: 0020-7136; 1097-0215
• DOI: 10.1002/ijc.21914

Nasopharyngeal carcinoma (NPC) is the most prevalent ENT‐tumour in Indonesia. We investigated the primary diagnostic value of Epstein‐Barr virus (EBV) DNA load and mRNA detection in noninvasive nasopharyngeal (NP) brushings, obtained prospectively from consecutive Indonesian ENT‐patients with suspected NPC (N = 106) and controls. A subsequent routine NP biopsy was taken for pathological examination and EBER‐RISH, yielding 85 confirmed NPC and 21 non‐NPC tumour patients. EBV DNA and human DNA load were quantified by real‐time PCR. NP brushings from NPC patients contained extremely high EBV DNA loads compared to the 88 non‐NPC controls (p < 0.0001). Using mean EBV DNA load in controls plus 3 SD as cut‐off value, specificity, sensitivity, positive and negative predictive values were 98, 90, 97 and 91%, respectively. Epstein‐Barr nuclear antigen 1 (EBNA1) and the carcinoma‐specific BARF1 mRNA were detected by nucleic acid sequence based amplification and found in 86 and 74% of NP brushings, confirming NPC tumour cell presence. EBV RNA positivity was even higher in fresh samples stored at −80°C until RNA expression analyses (88% for both EBNA1 and BARF1). EBV RNA‐negative NP brushings from proven NPC cases had the lowest EBV DNA loads, indicating erroneous sampling. No EBV mRNA was detected in NP brushings from healthy donors and non‐NPC patients. In conclusion, EBV DNA load measurement combined with detection of BARF1 mRNA in simple NP brushings allows noninvasive NPC diagnosis. It reflects carcinoma‐specific EBV involvement at the anatomical site of tumour development and reduces the need for invasive biopsies. This procedure may be useful for confirmatory diagnosis in large serological NPC screening programs and has potential as prognostic tool. © 2006 Wiley‐Liss, Inc.