Immunoassay for LMP1 in nasopharyngeal tissue based on surface-enhanced Raman scattering

Int J Nanomedicine. 2012:7:73-82. doi: 10.2147/IJN.S26854. Epub 2011 Dec 30.

Abstract

Background: Previous studies have shown that Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP1) is closely associated with the occurrence and development of nasopharyngeal carcinoma, and can be used as a tumor marker in screening for the disease. Here we report a new methodology based on highly specific and sensitive surface-enhanced Raman scattering (SERS) technology to detect LMP1 in nasopharyngeal tissue sections directly with no need of tedious procedures as with conventional immunohistochemistry methods.

Methods: LMP1-functionalized 4-mercaptobenzoic acid (4-MBA)-labeled Au/Ag core-shell bimetallic nanoparticles were prepared first and then applied for analyzing LMP1 in formalin-fixed paraffin-embedded nasopharyngeal tissue sections obtained from 34 cancer patients and 20 healthy controls. SERS spectra were acquired from a 25 × 25 spot square area on each tissue section and used to generate SERS images.

Results: Data from SERS spectra and images show that this new SERS-based immunoassay detected LMP1 in formalin-fixed paraffin-embedded nasopharyngeal tissue sections with high sensitivity and specificity. The results from the new LMP1-SERS probe method are superior to those of conventional immunohistochemistry staining for LMP1, and in excellent agreement with those of in situ hybridization for EBV-encoded small RNA (EBER).

Conclusion: This new SERS technique has the potential to be developed into a new clinical tool for detection and differential diagnosis of nasopharyngeal carcinoma as well as for predicting metastasis and immune-targeted treatment of nasopharyngeal carcinoma.

Keywords: LMP1; immunoassay; immunohistochemistry; in situ hybridization; nasopharyngeal carcinoma; surface-enhanced Raman scattering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Case-Control Studies
  • Epstein-Barr Virus Infections / virology
  • Female
  • Humans
  • Immunoassay / methods*
  • Immunohistochemistry
  • In Situ Hybridization
  • Least-Squares Analysis
  • Male
  • Metal Nanoparticles / chemistry
  • Microscopy, Electron, Transmission
  • Middle Aged
  • Nasopharyngeal Neoplasms / chemistry*
  • Nasopharyngeal Neoplasms / virology
  • Nasopharynx / chemistry
  • Nasopharynx / virology
  • Sensitivity and Specificity
  • Spectrum Analysis, Raman / instrumentation
  • Spectrum Analysis, Raman / methods*
  • Viral Matrix Proteins / analysis*
  • Viral Matrix Proteins / chemistry

Substances

  • EBV-associated membrane antigen, Epstein-Barr virus
  • Viral Matrix Proteins