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Detection and typing of herpes simplex viruses by using recombinant immunoglobulin fragments produced in bacteria

Academic Article
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Overview

authors

  • Cattani, P.
  • Rossolini, G. M.
  • Cresti, S.
  • Santangelo, R.
  • Burton, Dennis
  • Williamson, R. A.
  • Sanna, Pietro
  • Fadda, G.

publication date

  • June 1997

journal

  • Journal of Clinical Microbiology  Journal

abstract

  • Thirty-seven bacterial clones producing human recombinant monoclonal antibody Fab fragments (rFabs) reactive to herpes simplex virus (HSV) antigens were selected from a human combinatorial antibody library constructed in a phage-display vector by a panning procedure against an HSV lysate. Thirty-four of the HSV-specific rFabs were able to specifically recognize HSV-infected cells in indirect immunofluorescence (IF) assays; of these, 25 recognized cells infected by either HSV type 1 (HSV-1) or HSV-2, while 9 recognized only HSV-1-infected cells. One HSV type-common rFab (rFab H37) and one HSV-1-specific rFab (rFab H85) were further evaluated as reagents for viral detection and typing by IF staining in 134 HSV-positive (72 HSV-1 and 62 HSV-2) viral cultures from clinical specimens. The results obtained with these two rFabs were fully consistent with those obtained with a commercial preparation of fluorescein-labeled anti-HSV type-specific murine monoclonal antibodies. The detection sensitivity with the type-common rFab in indirect IF assays was higher overall than that provided by the type-specific murine monoclonal antibodies. Preparations of rFabs suitable for IF staining can be easily and inexpensively obtained in a clinical microbiology laboratory from Escherichia coli cultures. Similar HSV-specific rFabs, therefore, could be advantageous for in vitro diagnostic purposes.

subject areas

  • Animals
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral
  • Base Sequence
  • Cercopithecus aethiops
  • Escherichia coli
  • Fluorescent Antibody Technique, Indirect
  • Genetic Vectors
  • Herpesvirus 1, Human
  • Herpesvirus 2, Human
  • Humans
  • Immunoglobulin Fab Fragments
  • Molecular Sequence Data
  • Peptide Library
  • Recombinant Fusion Proteins
  • Sequence Analysis, DNA
  • Simplexvirus
  • Vero Cells
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Identity

PubMed Central ID

  • PMC229775

International Standard Serial Number (ISSN)

  • 0095-1137

PubMed ID

  • 9163470
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Additional Document Info

start page

  • 1504

end page

  • 1509

volume

  • 35

issue

  • 6

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