The application of the EBV-encoded nuclear antigen 1-targeting bioprobe in nasopharyngeal carcinoma diagnosis

Objectives:
We aim to (1) explore the potential application of our EBNA1 bioprobe L2P4 in detecting EBV in the plasma samples of NPC patients, and (2) determine whether L2P4 can be used as a rapid staining agent in FFPE tissues of NPC.

Hypothesis to be tested:
1. L2P4 can detect the presence of EBV in the plasma samples of NPC patients,

2. L2P4 can be used as a rapid staining agent of EBV with sensitivity like EBER hybridization in FFPE tissues of NPC.

Design and subjects:
The L2P4-induced EBNA1 aggregation (reflected by turbidity) will be measured in plasma samples in NPC patients versus healthy controls, and we will also determine whether the L2P4-induced aggregation levels can reflect the treatment outcome. L2P4 will be used as a dye to stain the presence of EBV in the FFPE tumor tissues of NPC patients.

Study instruments:
Measurement of turbidity by spectrophotometry, simple FFPE staining test with L2P4, qPCR, ELISA, and RNA hybridization.

Interventions:
N/A

Main outcome measures:
Sensitivity and specificity of using L2P4 to detect EBV in NPC plasma and FFPE tumor tissue samples.

Data analysis:
Receiver operating characteristic curve analysis will be performed to quantify the diagnostic potential of our EBNA1 detection methods.

Expected results:
The L2P4-induced EBNA1 aggregation can be used for detection of NPC using plasma samples, this assay in use with plasma EBV DNA qPCR can further enhance diagnostic sensitivity of detecting NPC. L2P4 can be also used as a rapid staining agent in FFPE tissues of NPC.