Production of recombinant gG-1 protein of herpes simplex virus type 1 in a prokaryotic system in order to develop a type-specific enzyme-linked immunosorbent assay kit.
The herpes simplex viruses are important causes of disease worldwide. Herpes simplex virus type 1 (HSV-1) is the primary cause of oral-facial and pharyngeal infections and may cause herpetic whitlow, eye infections as well as severe and sometimes dangerous infections of the eyes and brain. HSV-1 also accounts for 10-15% of all genital herpetic infections. Therefore, laboratory diagnosis of this virus and development of diagnostic serological techniques for HSV-1 is of particular importance. In the present study, pTrc His2A-gG1 plasmid, containing the full-length glycoprotein G (gG) protein, was produced in a prokaryotic system for the first time. Upon confirmation of a 37-kDa gG-1 protein production in a prokaryotic system based on western blotting and monoclonal antibodies, the protein was produced at a large scale and purified by ion-exchange chromatography using DEAE-sepharose. An HSV-1 type-specific diagnostic kit was designed and developed and the specificity and sensitivity of this kit were demonstrated to be 89.5% and 100%, respectively, as compared with a commercially available kit. A significant correlation was shown between the developed kit and the commercial kit.
Zandi
Keivan
K
Department of Microbiology, Faculty of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran. keivanzandi@yahoo.com
Roostaee
Mohammad Hassan
MH
Sadeghizadeh
Majid
M
Rasaee
Mohammad Javad
MJ
Sajedi
Reza Hassan
RH
Soleimanjahi
Hoorieh
H
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006561:Herpes Simplex / Q000175:diagnosis*; D018259:Herpesvirus 1, Human / Q000302:isolation & purification*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C050591:glycoprotein gG-1, herpes simplex virus type 1