Epstein-Barr Virus DNA Load in Nasopharyngeal Brushings and Whole Blood in Nasopharyngeal Carcinoma Patients before and after Treatment

Abstract

Purpose: Nasopharyngeal carcinoma (NPC) is consistently associated with Epstein-Barr virus (EBV) and highly prevalent in Indonesia. EBV-DNA load can be used for early diagnosis and may have prognostic value. In this study, EBV-DNA load was evaluated in minimal invasive nasopharyngeal (NP) brushings and whole blood for initial diagnosis and therapy assessment against the standard-of-care diagnosis by biopsy with EBV-RISH and standard EBV-IgA serology.Experimental Design: NP brushings and blood samples were collected from 289 consecutive ENT patients suspected of NPCs and 53 local healthy controls. EBV-DNA load was quantified by real-time PCR and serology by peptide-based EBV-IgA ELISA. Tissue biopsies were examined by routine histochemistry and by EBER RNA in situ hybridization.Results: Repeated NP brushing was well tolerated by patients and revealed high viral load in the 228 NPC cases at diagnosis than 61 non-NPC cancer cases and healthy controls (P < 0.001). The diagnostic value of EBV-DNA load in blood and EBV-IgA serology was inferior to the NP brush results. The level of EBV-DNA load in brushes of patients with NPC was not related to T, N, or M stage, whereas elevated EBV-DNA load in blood correlated with N and M stage. EBV-DNA levels in brushings and whole blood showed a significant reduction at 2 months after treatment (P ¼ 0.001 and P ¼ 0.005, respectively), which was not reflected in EBV-IgA serology.Conclusions: NP brush sampling combined with EBV-DNA load analysis is a minimal invasive and welltolerated diagnostic procedure, suited for initial diagnosis and follow-up monitoring of NPCs.

The prognostic value of IgA/[EBNA1+VCA-p18] on survival of nasopharyngeal cancer patients

“…Post-treatment IgA-EBV serology failed to determine prognostic difference among NPC patients. This is consistent with the previous studies done by Adham et al (2013) and Ai et al (2013). Direct quantification of viral EBV DNA, therefore, might lend a hand to support the notion that viral reactivation play important role in disease progression and survival (Hassen et al, 2011;Wang et al, 2013;Yip et al, 2014).…”

“…Similar findings was reported by Adham et al (2013) among NPC patients in Indonesia. Adham et al (2013) reported that there was no significant reduction at 2-months post-treatment of IgA EBV either IgA EBNA1 or IgA VCA-p18.…”

“…Direct quantification of viral EBV DNA, therefore, might lend a hand to support the notion that viral reactivation play important role in disease progression and survival (Hassen et al, 2011;Wang et al, 2013;Yip et al, 2014). EBV DNA load from nasopharyngeal brushings and whole blood showed significant reduction at 2-month after treatment, which was not reflected by EBV-IgA serology (Adham et al, 2013). …”

Circulating Epstein–Barr virus microRNAs miR‐BART7 and miR‐BART13 as biomarkers for nasopharyngeal carcinoma diagnosis and treatment

“…Furthermore, EBV DNA levels return to normal levels after successful radiotherapy; persistently raised EBV DNA levels may indicate a poor outcome after therapy. 49,50 While there is still no direct evidence, it has been suggested that the source of plasma EBV DNA is apoptotic cancer cells which are shed into the blood stream or circulating cancer cells as they migrate to distant sites to cause metastasis. 27 While the plasma EBV DNA which originates from apoptotic or detached circulating tumor cells is a valuable marker for NPC, it is possible that new EBV biomarkers which are associated with actively growing cancer cells may provide a new approach for evaluation of the presence of NPC cells.…”

Levels of plasma Epstein-Barr virus DNA prior and subsequent to treatment predicts the prognosis of nasopharyngeal carcinoma

“…EBV-DNA may be detected in NPC cells (5,6), and cell-free EBV-DNA may be detected in the plasma of patients with NPC (7)(8)(9)(10)(11)(12)(13). Furthermore, the levels of plasma EBV-DNA in recently diagnosed NPC patients have been significantly correlated with tumor volume (14,15), response to treatment (16), tumor clearance (17,18) and tumor recurrence (19)(20)(21).…”