Production of a fragment of glycoprotein G of herpes simplex virus type 2 and evaluation of its diagnostic potential

Singapore Med J. 2015 Jun;56(6):346-52. doi: 10.11622/smedj.2014197.

Abstract

Introduction: Herpes simplex virus type 2 (HSV-2) is the most common cause of genital herpes. Glycoprotein G (gG) is a prototype antigen for type-specific serodiagnosis distinguishing between HSV type 1 (HSV-1) and HSV-2 infections. As immunological diagnosis kits for accurate differentiation between HSV-1 and HSV-2 antibodies can be expensive, there is a need to develop a convenient, sensitive, specific and cost-effective serodiagnostic kit.

Methods: We successfully expressed a fragment of gG comprising residues 321-580 of HSV-2 with histidine tag (gG(321-580His)) in a Bac-to-Bac baculovirus expression system, which had an antigenicity similar to its native counterpart. An indirect enzyme-linked immunosorbent assay (ELISA) was developed using gG(321-580His) as the diagnostic antigen and evaluated by comparison with a commercial HerpeSelect 2 ELISA immunoglobulin G kit as reference.

Results: In testing 318 field serum samples, the diagnostic relative sensitivity and specificity of the developed gG(321-580His)-ELISA test in qualitative comparison with the commercial kit were 93.81% and 96.74%, respectively, and the accuracy was 94.65%.

Conclusion: The study indicates that gG(321-580His) has a high diagnostic potential for HSV-2 virus serodiagnosis in humans.

Keywords: baculovirus expression system; glycoprotein G; herpes simplex virus type 2; immunodominant fragment; indirect enzyme-linked immunosorbent assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antibodies, Viral / blood
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Herpes Genitalis / diagnosis*
  • Herpes Genitalis / virology
  • Herpes Simplex / diagnosis*
  • Herpes Simplex / virology
  • Herpesvirus 1, Human / isolation & purification
  • Herpesvirus 2, Human / isolation & purification*
  • Humans
  • Immunoglobulin G / chemistry*
  • Male
  • Polymerase Chain Reaction
  • Reagent Kits, Diagnostic
  • Reference Values
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Serologic Tests / methods*

Substances

  • Antibodies, Viral
  • Immunoglobulin G
  • Reagent Kits, Diagnostic